# Patent

## Title
Nucleic acid and corresponding protein entitled 184P1E2 useful in treatment and detection of cancer

## Abstract

A novel gene (designated 184P1E2) and its encoded protein, and variants thereof, are described wherein 184P1E2 exhibits tissue specific expression in normal adult tissue, and is aberrantly expressed in the cancers listed in Table I. Consequently, 184P1E2 provides a diagnostic, prognostic, prophylactic and/or therapeutic target for cancer. The 184P1E2 gene or fragment thereof, or its encoded protein, or variants thereof, or a fragment thereof, can be used to elicit a humoral or cellular immune response; antibodies or T cells reactive with 184P1E2 can be used in active or passive immunization.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. Ser. No. 11/368,284 filed Mar. 2, 2006, now U.S. Pat. No. 7,592,149, which is a division of U.S. Ser. No. 10/120,901 filed Apr. 9, 2002, now U.S. Pat. No. 7,135,549, issued Nov. 14, 2006, which claims priority from U.S. Ser. No. 60/282,739 filed Apr. 10, 2001, and U.S. Ser. No. 60/286,630, filed Apr. 25, 2001. The contents of these applications are hereby incorporated by reference herein in their entirety.


SUBMISSION ON COMPACT DISC
The content of the following submission on compact discs is incorporated herein by reference in its entirety: A compact disc copy of the Sequence Listing (COPY 1) (file name: 511582006701), date recorded: Apr. 5, 2006, size: 2,523,136 bytes ); a duplicate compact disc copy of the Sequence Listing (COPY 2) (file name: 511582006701), date recorded: Apr. 5, 2006, size: 2,523,136 bytes ); and a computer readable form copy of the Sequence Listing (CRF COPY) (file name: 511582006701), date recorded: Apr. 5, 2006, size: 2,523,136 bytes).
FIELD OF THE INVENTION
The invention described herein relates to a gene and its encoded protein, termed 184P1E2, expressed in certain cancers, and to diagnostic and therapeutic methods and compositions useful in the management of cancers that express 184P1E2.
BACKGROUND OF THE INVENTION
Cancer is the second leading cause of human death next to coronary disease. Worldwide, millions of people die from cancer every year. In the United States alone, as reported by the American Cancer Society, cancer causes the death of well over a half-million people annually, with over 1.2 million new cases diagnosed per year. While deaths from heart disease have been declining significantly, those resulting from cancer generally are on the rise. In the early part of the next century, cancer is predicted to become the leading cause of death.
Worldwide, several cancers stand out as the leading killers. In particular, carcinomas of the lung, prostate, breast, colon, pancreas, and ovary represent the primary causes of cancer death. These and virtually all other carcinomas share a common lethal feature. With very few exceptions, metastatic disease from a carcinoma is fatal. Moreover, even for those cancer patients who initially survive their primary cancers, common experience has shown that their lives are dramatically altered. Many cancer patients experience strong anxieties driven by the awareness of the potential for recurrence or treatment failure. Many cancer patients experience physical debilitations following treatment. Furthermore, many cancer patients experience a recurrence.
Worldwide, prostate cancer is the fourth most prevalent cancer in men. In North America and Northern Europe, it is by far the most common cancer in males and is the second leading cause of cancer death in men. In the United States alone, well over 30,000 men die annually of this disease—second only to lung cancer. Despite the magnitude of these figures, there is still no effective treatment for metastatic prostate cancer. Surgical prostatectomy, radiation therapy, hormone ablation therapy, surgical castration and chemotherapy continue to be the main treatment modalities. Unfortunately, these treatments are ineffective for many and are often associated with undesirable consequences.
On the diagnostic front, the lack of a prostate tumor marker that can accurately detect early-stage, localized tumors remains a significant limitation in the diagnosis and management of this disease. Although the serum prostate specific antigen (PSA) assay has been a very useful tool, however its specificity and general utility is widely regarded as lacking in several important respects.
Progress in identifying additional specific markers for prostate cancer has been improved by the generation of prostate cancer xenografts that can recapitulate different stages of the disease in mice. The LAPC (Los Angeles Prostate Cancer) xenografts are prostate cancer xenografts that have survived passage in severe combined immune deficient (SCID) mice and have exhibited the capacity to mimic the transition from androgen dependence to androgen independence (Klein et al., 1997, Nat. Med. 3:402). More recently identified prostate cancer markers include PCTA-1 (Su et al., 1996, Proc. Natl. Acad. Sci. USA 93: 7252), prostate-specific membrane (PSM) antigen (Pinto et al., Clin Cancer Res 1996 Sep. 2 (9): 1445-51), STEAP (Hubert, et al., Proc Natl Acad Sci USA. 1999 Dec. 7; 96(25): 14523-8) and prostate stem cell antigen (PSCA) (Reiter et al., 1998, Proc. Natl. Acad. Sci. USA 95: 1735).
While previously identified markers such as PSA, PSM, PCTA and PSCA have facilitated efforts to diagnose and treat prostate cancer, there is need for the identification of additional markers and therapeutic targets for prostate and related cancers in order to further improve diagnosis and therapy.
Renal cell carcinoma (RCC) accounts for approximately 3 percent of adult malignancies. Once adenomas reach a diameter of 2 to 3 cm, malignant potential exists. In the adult, the two principal malignant renal tumors are renal cell adenocarcinoma and transitional cell carcinoma of the renal pelvis or ureter. The incidence of renal cell adenocarcinoma is estimated at more than 29,000 cases in the United States, and more than 11,600 patients died of this disease in 1998. Transitional cell carcinoma is less frequent, with an incidence of approximately 500 cases per year in the United States.
Surgery has been the primary therapy for renal cell adenocarcinoma for many decades. Until recently, metastatic disease has been refractory to any systemic therapy. With recent developments in systemic therapies, particularly immunotherapies, metastatic renal cell carcinoma may be approached aggressively in appropriate patients with a possibility of durable responses. Nevertheless, there is a remaining need for effective therapies for these patients.
Of all new cases of cancer in the United States, bladder cancer represents approximately 5 percent in men (fifth most common neoplasm) and 3 percent in women (eighth most common neoplasm). The incidence is increasing slowly, concurrent with an increasing older population. In 1998, there was an estimated 54,500 cases, including 39,500 in men and 15,000 in women. The age-adjusted incidence in the United States is 32 per 100,000 for men and 8 per 100,000 in women. The historic male/female ratio of 3:1 may be decreasing related to smoking patterns in women. There were an estimated 11,000 deaths from bladder cancer in 1998 (7,800 in men and 3,900 in women). Bladder cancer incidence and mortality strongly increase with age and will be an increasing problem as the population becomes more elderly.
Most bladder cancers recur in the bladder. Bladder cancer is managed with a combination of transurethral resection of the bladder (TUR) and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer points out the limitations of TUR. Most muscle-invasive cancers are not cured by TUR alone. Radical cystectomy and urinary diversion is the most effective means to eliminate the cancer but carry an undeniable impact on urinary and sexual function. There continues to be a significant need for treatment modalities that are beneficial for bladder cancer patients.
An estimated 130,200 cases of colorectal cancer occurred in 2000 in the United States, including 93,800 cases of colon cancer and 36,400 of rectal cancer. Colorectal cancers are the third most common cancers in men and women. Incidence rates declined significantly during 1992-1996 (−2.1% per year). Research suggests that these declines have been due to increased screening and polyp removal, preventing progression of polyps to invasive cancers. There were an estimated 56,300 deaths (47,700 from colon cancer, 8,600 from rectal cancer) in 2000, accounting for about 11% of all U.S. cancer deaths.
At present, surgery is the most common form of therapy for colorectal cancer, and for cancers that have not spread, it is frequently curative. Chemotherapy, or chemotherapy plus radiation, is given before or after surgery to most patients whose cancer has deeply perforated the bowel wall or has spread to the lymph nodes. A permanent colostomy (creation of an abdominal opening for elimination of body wastes) is occasionally needed for colon cancer and is infrequently required for rectal cancer. There continues to be a need for effective diagnostic and treatment modalities for colorectal cancer.
There were an estimated 164,100 new cases of lung and bronchial cancer in 2000, accounting for 14% of all U.S. cancer diagnoses. The incidence rate of lung and bronchial cancer is declining significantly in men, from a high of 86.5 per 100,000 in 1984 to 70.0 in 1996. In the 1990s, the rate of increase among women began to slow. In 1996, the incidence rate in women was 42.3 per 100,000.
Lung and bronchial cancer caused an estimated 156,900 deaths in 2000, accounting for 28% of all cancer deaths. During 1992-1996, mortality from lung cancer declined significantly among men (−1.7% per year) while rates for women were still significantly increasing (0.9% per year). Since 1987, more women have died each year of lung cancer than breast cancer, which, for over 40 years, was the major cause of cancer death in women. Decreasing lung cancer incidence and mortality rates most likely resulted from decreased smoking rates over the previous 30 years; however, decreasing smoking patterns among women lag behind those of men. Of concern, although the declines in adult tobacco use have slowed, tobacco use in youth is increasing again.
Treatment options for lung and bronchial cancer are determined by the type and stage of the cancer and include surgery, radiation therapy, and chemotherapy. For many localized cancers, surgery is usually the treatment of choice. Because the disease has usually spread by the time it is discovered, radiation therapy and chemotherapy are often needed in combination with surgery. Chemotherapy alone or combined with radiation is the treatment of choice for small cell lung cancer; on this regimen, a large percentage of patients experience remission, which in some cases is long lasting. There is however, an ongoing need for effective treatment and diagnostic approaches for lung and bronchial cancers.
An estimated 182,800 new invasive cases of breast cancer were expected to occur among women in the United States during 2000. Additionally, about 1,400 new cases of breast cancer were expected to be diagnosed in men in 2000. After increasing about 4% per year in the 1980s, breast cancer incidence rates in women have leveled off in the 1990s to about 110.6 cases per 100,000.
In the U.S. alone, there were an estimated 41,200 deaths (40,800 women, 400 men) in 2000 due to breast cancer. Breast cancer ranks second among cancer deaths in women. According to the most recent data, mortality rates declined significantly during 1992-1996 with the largest decreases in younger women, both white and black. These decreases were probably the result of earlier detection and improved treatment.
Taking into account the medical circumstances and the patient's preferences, treatment of breast cancer may involve lumpectomy (local removal of the tumor) and removal of the lymph nodes under the arm; mastectomy (surgical removal of the breast) and removal of the lymph nodes under the arm; radiation therapy; chemotherapy; or hormone therapy. Often, two or more methods are used in combination. Numerous studies have shown that, for early stage disease, long-term survival rates after lumpectomy plus radiotherapy are similar to survival rates after modified radical mastectomy. Significant advances in reconstruction techniques provide several options for breast reconstruction after mastectomy. Recently, such reconstruction has been done at the same time as the mastectomy.
Local excision of ductal carcinoma in situ (DCIS) with adequate amounts of surrounding normal breast tissue may prevent the local recurrence of the DCIS. Radiation to the breast and/or tamoxifen may reduce the chance of DCIS occurring in the remaining breast tissue. This is important because DCIS, if left untreated, may develop into invasive breast cancer. Nevertheless, there are serious side effects or sequelae to these treatments. There is, therefore, a need for efficacious breast cancer treatments.
There were an estimated 23,100 new cases of ovarian cancer in the United States in 2000. It accounts for 4% of all cancers among women and ranks second among gynecologic cancers. During 1992-1996, ovarian cancer incidence rates were significantly declining. Consequent to ovarian cancer, there were an estimated 14,000 deaths in 2000. Ovarian cancer causes more deaths than any other cancer of the female reproductive system.
Surgery, radiation therapy, and chemotherapy are treatment options for ovarian cancer. Surgery usually includes the removal of one or both ovaries, the fallopian tubes (salpingo-oophorectomy), and the uterus (hysterectomy). In some very early tumors, only the involved ovary will be removed, especially in young women who wish to have children. In advanced disease, an attempt is made to remove all intra-abdominal disease to enhance the effect of chemotherapy. There continues to be an important need for effective treatment options for ovarian cancer.
There were an estimated 28,300 new cases of pancreatic cancer in the United States in 2000. Over the past 20 years, rates of pancreatic cancer have declined in men. Rates among women have remained approximately constant but may be beginning to decline. Pancreatic cancer caused an estimated 28,200 deaths in 2000 in the United States. Over the past 20 years, there has been a slight but significant decrease in mortality rates among men (about −0.9% per year) while rates have increased slightly among women.
Surgery, radiation therapy, and chemotherapy are treatment options for pancreatic cancer. These treatment options can extend survival and/or relieve symptoms in many patients but are not likely to produce a cure for most. There is a significant need for additional therapeutic and diagnostic options for pancreatic cancer.
SUMMARY OF THE INVENTION
The present invention relates to a gene, designated 184P1E2, that has now been found to be over-expressed in the cancer(s) listed in Table I. Northern blot expression analysis of 184P1E2 gene expression in normal tissues shows a restricted expression pattern in adult tissues. The nucleotide (FIG. 2) and amino acid (FIG. 2, and FIG. 3) sequences of 184P1E2 are provided. The tissue-related profile of 184P1E2 in normal adult tissues, combined with the over-expression observed in the tissues listed in Table I, shows that 184P1E2 is aberrantly over-expressed in at least some cancers, and thus serves as a useful diagnostic, prophylactic, prognostic, and/or therapeutic target for cancers of the tissue(s) such as those listed in Table I.
The invention provides polynucleotides corresponding or complementary to all or part of the 184P1E2 genes, mRNAs, and/or coding sequences, preferably in isolated form, including polynucleotides encoding 184P1E2-related proteins and fragments of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 contiguous amino acids; at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 or more than 100 contiguous amino acids of a 184P1E2-related protein, as well as the peptides/proteins themselves; DNA, RNA, DNA/RNA hybrids, and related molecules, polynucleotides or oligonucleotides complementary or having at least a 90% homology to the 184P1E2 genes or mRNA sequences or parts thereof, and polynucleotides or oligonucleotides that hybridize to the 184P1E2 genes, mRNAs, or to 184P1E2-encoding polynucleotides. Also provided are means for isolating cDNAs and the genes encoding 184P1E2. Recombinant DNA molecules containing 184P1E2 polynucleotides, cells transformed or transduced with such molecules, and host-vector systems for the expression of 184P1E2 gene products are also provided. The invention further provides antibodies that bind to 184P1E2 proteins and polypeptide fragments thereof, including polyclonal and monoclonal antibodies, murine and other mammalian antibodies, chimeric antibodies, humanized and fully human antibodies, and antibodies labeled with a detectable marker or therapeutic agent. In certain embodiments there is a proviso that the entire nucleic acid sequence of FIG. 2 is not encoded and/or the entire amino acid sequence of FIG. 2 is not prepared. In certain embodiments, the entire nucleic acid sequence of FIG. 2 is encoded and/or the entire amino acid sequence of FIG. 2 is prepared, either of which are in respective human unit dose forms.
The invention further provides methods for detecting the presence and status of 184P1E2 polynucleotides and proteins in various biological samples, as well as methods for identifying cells that express 184P1E2. A typical embodiment of this invention provides methods for monitoring 184P1E2 gene products in a tissue or hematology sample having or suspected of having some form of growth dysregulation such as cancer.
The invention further provides various immunogenic or therapeutic compositions and strategies for treating cancers that express 184P1E2 such as cancers of tissues listed in Table I, including therapies aimed at inhibiting the transcription, translation, processing or function of 184P1E2 as well as cancer vaccines. In one aspect, the invention provides compositions, and methods comprising them, for treating a cancer that expresses 184P1E2 in a human subject wherein the composition comprises a carrier suitable for human use and a human unit dose of one or more than one agent that inhibits the production or function of 184P1E2. Preferably, the carrier is a uniquely human carrier. In another aspect of the invention, the agent is a moiety that is immunoreactive with 184P1E2 protein. Non-limiting examples of such moieties include, but are not limited to, antibodies (such as single chain, monoclonal, polyclonal, humanized, chimeric, or human antibodies), functional equivalents thereof (whether naturally occurring or synthetic), and combinations thereof. The antibodies can be conjugated to a diagnostic or therapeutic moiety. In another aspect, the agent is a small molecule as defined herein.
In another aspect, the agent comprises one or more than one peptide which comprises a cytotoxic T lymphocyte (CTL) epitope that binds an HLA class I molecule in a human to elicit a CTL response to 184P1E2 and/or one or more than one peptide which comprises a helper T lymphocyte (HTL) epitope which binds an HLA class II molecule in a human to elicit an HTL response. The peptides of the invention may be on the same or on one or more separate polypeptide molecules. In a further aspect of the invention, the agent comprises one or more than one nucleic acid molecule that expresses one or more than one of the CTL or HTL response stimulating peptides as described above. In yet another aspect of the invention, the one or more than one nucleic acid molecule may express a moiety that is immunologically reactive with 184P1E2 as described above. The one or more than one nucleic acid molecule may also be, or encodes, a molecule that inhibits production of 184P1E2. Non-limiting examples of such molecules include, but are not limited to, those complementary to a nucleotide sequence essential for production of 184P1E2 (e.g. antisense sequences or molecules that form a triple helix with a nucleotide double helix essential for 184P1E2 production) or a ribozyme effective to lyse 184P1E2 mRNA.
Note that to determine the starting position of any peptide set forth in Tables V-XVII and XXII to LI (collectively HLA Peptide Tables) respective to its parental protein, e.g., variant 1, variant 2, etc., reference is made to three factors: the particular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides in Table LII. Generally, a unique Search Peptide is used to obtain HLA peptides of a particular for a particular variant. The position of each Search Peptide relative to its respective parent molecule is listed in Table LII. Accordingly, if a Search Peptide begins at position “X”, one must add the value “X−1” to each position in Tables V-XVIII and XXII to LI to obtain the actual position of the HLA peptides in their parental molecule. For example, if a particular Search Peptide begins at position 150 of its parental molecule, one must add 150−1, i.e., 149 to each HLA peptide amino acid position to calculate the position of that amino acid in the parent molecule.
One embodiment of the invention comprises an HLA peptide, that occurs at least twice in Tables V-XVIII and XXI to LI collectively, or an oligonucleotide that encodes the HLA peptide. Another embodiment of the invention comprises an HLA peptide that occurs at least once in Tables V-XVIII and at least once in tables XXII to LI, or an oligonucleotide that encodes the HLA peptide.
Another embodiment of the invention is antibody epitopes which comprise a peptide regions, or an oligonucleotide encoding the peptide region, that has one two, three, four, or five of the following characteristics:
i) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Hydrophilicity profile of FIG. 5;
ii) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or less than 0.5, 0.4, 0.3, 0.2, 0.1, or having a value equal to 0.0, in the Hydropathicity profile of FIG. 6;
iii) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Percent Accessible Residues profile of FIG. 7;
iv) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Average Flexibility profile of FIG. 8; or
v) a peptide region of at least 5 amino acids of a particular peptide of FIG. 3, in any whole number increment up to the full length of that protein in FIG. 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Beta-turn profile of FIG. 9.



BRIEF DESCRIPTION OF THE FIGURES
FIG. 1. The 184P1E2 SSH sequence (SEQ ID NO.: 1) of 132 nucleotides.
FIG. 2. The cDNA (SEQ ID. NO.: 2) and amino acid sequence (SEQ ID. NO.: 3) of 184P1E2 variant 1 (also called “184P1E2 v.1” or “184P1E2 variant 1”) is shown in FIG. 2A. The start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 4) and amino acid sequence (SEQ ID. NO.: 5) of 184P1E2 variant 2 (also called “184P1E2 v.2”) is shown in FIG. 2B. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 6) and amino acid sequence (SEQ ID. NO.: 7) of 184P1E2 variant 3 (also called “184P1E2 v.3”) is shown in FIG. 2C. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 8) and amino acid sequence (SEQ ID. NO.: 9) of 184P1E2 variant 4 (also called “184P1E2 v.4”) is shown in FIG. 2D. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 10) and amino acid sequence (SEQ ID. NO.: 11) of 184P1E2 variant 5 (also called “184P1E2 v.5”) is shown in FIG. 2E. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 12) and amino acid sequence (SEQ ID. NO.: 13) of 184P1E2 variant 6 (also called “184P1E2 v.6”) is shown in FIG. 2F. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 14) and amino acid sequence (SEQ ID. NO.: 15) of 184P1E2 variant 7 (also called “184P1E2 v.7”) is shown in FIG. 2G. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 16) and amino acid sequence (SEQ ID. NO.: 17) of 184P1E2 variant 8 (also called “184P1E2 v.8”) is shown in FIG. 2H. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 18) and amino acid sequence (SEQ ID. NO.: 19) of 184P1E2 variant 9 (also called “184P1E2 v.9”) is shown in FIG. 2I. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. The cDNA (SEQ ID. NO.: 20) and amino acid sequence (SEQ ID. NO.: 21) of 184P1E2 variant 10 (also called “184P1E2 v.10”) is shown in FIG. 2J. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 42-2036 including the stop codon. As used herein, a reference to 184P1E2 includes all variants thereof, including those shown in FIGS. 10 and 12.
FIG. 3. Amino acid sequence of 184P1E2 v.1 (SEQ ID. NO.: 3) is shown in FIG. 3A; it has 664 amino acids. The amino acid sequence of 184P1E2 v.2 (SEQ ID. NO.: 4) is shown in FIG. 3B; it has 664 amino acids. The amino acid sequence of 184P1E2 v.3 (SEQ ID. NO.: 5) is shown in FIG. 3C; it has 664 amino acids. As used herein, a reference to 184P1E2 includes all variants thereof, including those shown in FIG. 11.
FIG. 4. The nucleic acid sequence alignment of 184P1E2 v.1 (SEQ ID NO.: 2) with human peptidylarginine deiminase type III (SEQ ID NO.: 22) is shown in FIG. 4A. The nucleic acid differences are underlined. The amino acid sequence alignment of 184P1E2 v.1 (SEQ ID NO.: 3) with human peptidylarginine deiminase type III (SEQ ID NO.: 23) is shown in FIG. 4B. The amino acid variation at position 480 is underlined. The amino acid sequence alignment of 184P1E2 v.1 (SEQ ID NO.: 3) with mouse peptidylarginine deiminase type III (SEQ ID NO.: 24) is shown in FIG. 4C. The amino acid sequence alignment of 184P1E2 v.1 (SEQ ID NO.: 3) with Rat deiminase type III (SEQ ID NO.: 25) is shown in FIG. 4D.
FIG. 5. Hydrophilicity amino acid profile of 184P1E2 variant 1, determined by computer algorithm sequence analysis using the method of Hopp and Woods (Hopp T. P., Woods K. R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828) accessed on the Protscale website through the ExPasy molecular biology server.
FIG. 6. Hydropathicity amino acid profile of 184P1E2 variant 1, determined by computer algorithm sequence analysis using the method of Kyte and Doolittle (Kyte J., Doolittle R. F., 1982. J. Mol. Biol. 157:105-132) accessed on the ProtScale website through the ExPasy molecular biology server.
FIG. 7. Percent accessible residues amino acid profile of 184P1E2 variant 1, determined by computer algorithm sequence analysis using the method of Janin (Janin J., 1979 Nature 277:491-492) accessed on the ProtScale website through the ExPasy molecular biology server.
FIG. 8. Average flexibility amino acid profile of 184P1E2 variant 1, determined by computer algorithm sequence analysis using the method of Bhaskaran and Ponnuswamy (Bhaskaran R., and Ponnuswamy P. K., 1988. Int. J. Pept. Protein Res. 32:242-255) accessed on the ProtScale website through the ExPasy molecular biology server.
FIG. 9. Beta-turn amino acid profile of 184P1E2 variant 1, determined by computer algorithm sequence analysis using the method of Deleage and Roux (Deleage, G., Roux B. 1987 Protein Engineering 1:289-294) accessed on the ProtScale website through the ExPasy molecular biology server.
FIG. 10. Schematic display of nucleotide variants of 184P1E2. Variants 184P1E2 v.2 through v.10 are variants with single nucleotide variation. The black boxes show the same sequence as 184P1E2 v.1. The numbers correspond to those of 184P1E2 v.1. Single Nucleotide Polymorphisms (also called “SNPs”) are indicated above the boxes.
FIG. 11. Schematic display of protein variants of 184P1E2. Nucleotide variants 184P1E2 v.1, v.2 and v.3 in FIG. 10 code for protein variants 184P1E2 v.1, v.2 and v.3, respectively. Variants 184P1E2 v.4 through v.10 code the same protein as variant 184P1E2 v.1. Protein variants 184P1E2 v.2 and v.3 are variants with single amino acid variations. The black boxes show the same sequence as 184P1E2 v.1. The numbers correspond to those of 184P1E2 v.1. Single amino acid differences are indicated above the box.
FIG. 12. The exon composition of the original transcript, designated as 184P1E2 v.1.
FIG. 13. Secondary structure prediction for 184P1E2 (SEQ ID NO.: 3). The secondary structure of 184P1E2 variant 1 was predicted using the HNN—Hierarchical Neural Network method (Guermeur, 1997), accessed from the ExPasy molecular biology server. This method predicts the presence and location of alpha helices, extended strands, and random coils from the primary protein sequence. The percent of the protein in a given secondary structure is also listed.
FIG. 14. Expression of 184P1E2 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P1E2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 184P1E2 in bladder cancer pool. Expression of 184P1E2 is also detected in kidney cancer pool, lung cancer pool and cancer metastasis pool, but not in vital pool 1 and vital pool 2.
FIG. 15. Expression of 184P1E2 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 ug of mRNA/lane were probed with the 184P1E2 sequence. Size standards in kilobases (kb) are indicated on the side. Results show absence of expression of 184P1E2 in all 16 normal tissues tested.
FIG. 16. Expression of 184P1E2 in Patient Cancer Specimens and Normal Tissues. RNA was extracted from a pool of three bladder cancers (BCP), as well as from normal prostate (NP), normal bladder (NB), normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr) and normal ovary (NO). Northern blot with 10 μg of total RNAAane was probed with 184P1E2 sequence. Size standards in kilobases (kb) are indicated on the side. An approximately 4.5 kb 184P1E2 transcript was detected in the bladder cancer pool but not in the normal tissues tested.
FIG. 17. Expression of 184P1E2 in bladder cancer patient tissues. RNA was extracted from normal bladder (NB), bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), bladder cancer patient tumors (T) and their normal adjacent tissues (N). Northern blots with 10 ug of total RNA were probed with the 184P1E2 sequence. Size standards in kilobases are indicated on the side. Results show strong expression of 184P1E2 in patient bladder cancer tissues.
FIG. 18. Expression of 184P1E2 in lung cancer patient tissues. RNA was extracted from lung cancer cell lines (CL) (CALU-1, A427, NCI-H82, NCI-146), normal lung (N), lung cancer patient tumors (T) and their normal adjacent tissues (Nat). Northern blots with 10 ug of total RNA were probed with the 184P1E2 sequence. Size standards in kilobases are on the side. Results show strong expression of 184P1E2 in patient lung cancer tissues, but not in normal lung. A lower molecular weight transcript was also detected in the two cell lines CALU-1 and NCI-146.



DETAILED DESCRIPTION OF THE INVENTION
Outline of Sections
I.) Definitions
II.) 184P1E2 Polynucleotides
II.A.) Uses of 184P1E2 Polynucleotides
II.A.1.) Monitoring of Genetic Abnormalities
II.A.2.) Antisense Embodiments
II.A.3.) Primers and Primer Pairs
II.A.4.) Isolation of 184P1E2-Encoding Nucleic Acid Molecules
II.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems
III.) 184P1E2-related Proteins
III.A.) Motif-bearing Protein Embodiments
III.B.) Expression of 184P1E2-related Proteins
III.C.) Modifications of 184P1E2-related Proteins
III.D.) Uses of 184P1E2-related Proteins
IV.) 184P1E2 Antibodies
V.) 184P1E2 Cellular Immune Responses
VI.) 184P1E2 Transgenic Animals
VII.) Methods for the Detection of 184P1E2
VIII.) Methods for Monitoring the Status of 184P1E2-related Genes and Their Products
IX.) Identification of Molecules That Interact With 184P1E2
X.) Therapeutic Methods and Compositions
X.A.) Anti-Cancer Vaccines
X.B.) 184P1E2 as a Target for Antibody-Based Therapy
X.C.) 184P1E2 as a Target for Cellular Immune Responses
X.C.1. Minigene Vaccines
X.C.2. Combinations of CTL Peptides with Helper Peptides
X.C.3. Combinations of CTL Peptides with T Cell Priming Agents
X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or HTL Peptides
X.D.) Adoptive Immunotherapy
X.E.) Administration of Vaccines for Therapeutic or Prophylactic Purposes
XI.) Diagnostic and Prognostic Embodiments of 184P1E2.
XII.) Inhibition of 184P1E2 Protein Function
XII.A.) Inhibition of 184P1E2 With Intracellular Antibodies
XII.B.) Inhibition of 184P1E2 with Recombinant Proteins
XII.C.) Inhibition of 184P1E2 Transcription or Translation
XII.D.) General Considerations for Therapeutic Strategies
XIII.) KITS
I.) Definitions:
Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many of the techniques and procedures described or referenced herein are well understood and commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 2nd. edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and/or parameters unless otherwise noted.
The terms “advanced prostate cancer”, “locally advanced prostate cancer”, “advanced disease” and “locally advanced disease” mean prostate cancers that have extended through the prostate capsule, and are meant to include stage C disease under the American Urological Association (AUA) system, stage C1-C2 disease under the Whitmore-Jewett system, and stage T3-T4 and N+ disease under the TNM (tumor, node, metastasis) system. In general, surgery is not recommended for patients with locally advanced disease, and these patients have substantially less favorable outcomes compared to patients having clinically localized (organ-confined) prostate cancer. Locally advanced disease is clinically identified by palpable evidence of induration beyond the lateral border of the prostate, or asymmetry or induration above the prostate base. Locally advanced prostate cancer is presently diagnosed pathologically following radical prostatectomy if the tumor invades or penetrates the prostatic capsule, extends into the surgical margin, or invades the seminal vesicles.
“Altering the native glycosylation pattern” is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence 184P1E2 (either by removing the underlying glycosylation site or by deleting the glycosylation by chemical and/or enzymatic means), and/or adding one or more glycosylation sites that are not present in the native sequence 184P1E2. In addition, the phrase includes qualitative changes in the glycosylation of the native proteins, involving a change in the nature and proportions of the various carbohydrate moieties present.
The term “analog” refers to a molecule which is structurally similar or shares similar or corresponding attributes with another molecule (e.g. a 184P1E2-related protein). For example an analog of a 184P1E2 protein can be specifically bound by an antibody or T cell that specifically binds to 184P1E2.
The term “antibody” is used in the broadest sense. Therefore an “antibody” can be naturally occurring or man-made such as monoclonal antibodies produced by conventional hybridoma technology. Anti-184P1E2 antibodies comprise monoclonal and polyclonal antibodies as well as fragments containing the antigen-binding domain and/or one or more complementarity determining regions of these antibodies.
An “antibody fragment” is defined as at least a portion of the variable region of the immunoglobulin molecule that binds to its target, i.e., the antigen-binding region. In one embodiment it specifically covers single anti-184P1E2 antibodies and clones thereof (including agonist, antagonist and neutralizing antibodies) and anti-184P1E2 antibody compositions with polyepitopic specificity.
The term “codon optimized sequences” refers to nucleotide sequences that have been optimized for a particular host species by replacing any codons having a usage frequency of less than about 20%. Nucleotide sequences that have been optimized for expression in a given host species by elimination of spurious polyadenylation sequences, elimination of exon/intron splicing signals, elimination of transposon-like repeats and/or optimization of GC content in addition to codon optimization are referred to herein as an “expression enhanced sequences.”
The term “cytotoxic agent” refers to a substance that inhibits or prevents the expression activity of cells, function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof. Examples of cytotoxic agents include, but are not limited to maytansinoids, yttrium, bismuth, ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, alpha-sarcin, gelonin, mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, sapaonaria officinalis inhibitor, and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes such as At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32 and radioactive isotopes of Lu. Antibodies may also be conjugated to an anti-cancer pro-drug activating enzyme capable of converting the pro-drug to its active form.
The term “homolog” refers to a molecule which exhibits homology to another molecule, by for example, having sequences of chemical residues that are the same or similar at corresponding positions.
“Human Leukocyte Antigen” or “HLA” is a human class I or class II Major Histocompatibility Complex (MHC) protein (see, e.g., Stites, et al., IMMUNOLOGY, 8TH ED., Lange Publishing, Los Altos, Calif. (1994).
The terms “hybridize”, “hybridizing”, “hybridizes” and the like, used in the context of polynucleotides, are meant to refer to conventional hybridization conditions, preferably such as hybridization in 50% formamide/6×SSC/0.1% SDS/100 μg/ml ssDNA, in which temperatures for hybridization are above 37 degrees C. and temperatures for washing in 0.1×SSC/0.1% SDS are above 55 degrees C.
The phrases “isolated” or “biologically pure” refer to material which is substantially or essentially free from components which normally accompany the material as it is found in its native state. Thus, isolated peptides in accordance with the invention preferably do not contain materials normally associated with the peptides in their in situ environment. For example, a polynucleotide is said to be “isolated” when it is substantially separated from contaminant polynucleotides that correspond or are complementary to genes other than the 184P1E2 genes or that encode polypeptides other than 184P1E2 gene product or fragments thereof. A skilled artisan can readily employ nucleic acid isolation procedures to obtain an isolated 184P1E2 polynucleotide. A protein is said to be “isolated,” for example, when physical, mechanical or chemical methods are employed to remove the 184P1E2 proteins from cellular constituents that are normally associated with the protein. A skilled artisan can readily employ standard purification methods to obtain an isolated 184P1E2 protein. Alternatively, an isolated protein can be prepared by chemical means.
The term “mammal” refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses and humans. In one embodiment of the invention, the mammal is a mouse. In another embodiment of the invention, the mammal is a human.
The terms “metastatic prostate cancer” and “metastatic disease” mean prostate cancers that have spread to regional lymph nodes or to distant sites, and are meant to include stage D disease under the AUA system and stage T×N×M+under the TNM system. As is the case with locally advanced prostate cancer, surgery is generally not indicated for patients with metastatic disease, and hormonal (androgen ablation) therapy is a preferred treatment modality. Patients with metastatic prostate cancer eventually develop an androgen-refractory state within 12 to 18 months of treatment initiation. Approximately half of these androgen-refractory patients die within 6 months after developing that status. The most common site for prostate cancer metastasis is bone. Prostate cancer bone metastases are often osteoblastic rather than osteolytic (i.e., resulting in net bone formation). Bone metastases are found most frequently in the spine, followed by the femur, pelvis, rib cage, skull and humerus. Other common sites for metastasis include lymph nodes, lung, liver and brain. Metastatic prostate cancer is typically diagnosed by open or laparoscopic pelvic lymphadenectomy, whole body radionuclide scans, skeletal radiography, and/or bone lesion biopsy.
The term “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the antibodies comprising the population are identical except for possible naturally occurring mutations that are present in minor amounts.
A “motif”, as in biological motif of a 184P1E2-related protein, refers to any pattern of amino acids forming part of the primary sequence of a protein, that is associated with a particular function (e.g. protein-protein interaction, protein-DNA interaction, etc) or modification (e.g. that is phosphorylated, glycosylated or amidated), or localization (e.g. secretory sequence, nuclear localization sequence, etc.) or a sequence that is correlated with being immunogenic, either humorally or cellularly. A motif can be either contiguous or capable of being aligned to certain positions that are generally correlated with a certain function or property. In the context of HLA motifs, “motif” refers to the pattern of residues in a peptide of defined length, usually a peptide of from about 8 to about 13 amino acids for a class I HLA motif and from about 6 to about 25 amino acids for a class II HLA motif, which is recognized by a particular HLA molecule. Peptide motifs for HLA binding are typically different for each protein encoded by each human HLA allele and differ in the pattern of the primary and secondary anchor residues.
A “pharmaceutical excipient” comprises a material such as an adjuvant, a carrier, pH-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservative, and the like.
“Pharmaceutically acceptable” refers to a non-toxic, inert, and/or composition that is physiologically compatible with humans or other mammals.
The term “polynucleotide” means a polymeric form of nucleotides of at least 10 bases or base pairs in length, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide, and is meant to include single and double stranded forms of DNA and/or RNA. In the art, this term if often used interchangeably with “oligonucleotide”. A polynucleotide can comprise a nucleotide sequence disclosed herein wherein thymidine (T), as shown for example in FIG. 2, can also be uracil (U); this definition pertains to the differences between the chemical structures of DNA and RNA, in particular the observation that one of the four major bases in RNA is uracil (U) instead of thymidine (T).
The term “polypeptide” means a polymer of at least about 4, 5, 6, 7, or 8 amino acids. Throughout the specification, standard three letter or single letter designations for amino acids are used. In the art, this term is often used interchangeably with “peptide” or “protein”.
An HLA “primary anchor residue” is an amino acid at a specific position along a peptide sequence which is understood to provide a contact point between the immunogenic peptide and the HLA molecule. One to three, usually two, primary anchor residues within a peptide of defined length generally defines a “motif” for an immunogenic peptide. These residues are understood to fit in close contact with peptide binding groove of an HLA molecule, with their side chains buried in specific pockets of the binding groove. In one embodiment, for example, the primary anchor residues for an HLA class I molecule are located at position 2 (from the amino terminal position) and at the carboxyl terminal position of a 8, 9, 10, 11, or 12 residue peptide epitope in accordance with the invention. In another embodiment, for example, the primary anchor residues of a peptide that will bind an HLA class II molecule are spaced relative to each other, rather than to the termini of a peptide, where the peptide is generally of at least 9 amino acids in length. The primary anchor positions for each motif and supermotif are set forth in Table IV. For example, analog peptides can be created by altering the presence or absence of particular residues in the primary and/or secondary anchor positions shown in Table IV. Such analogs are used to modulate the binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supermotif.
A “recombinant” DNA or RNA molecule is a DNA or RNA molecule that has been subjected to molecular manipulation in vitro.
Non-limiting examples of small molecules include compounds that bind or interact with 184P1E2, ligands including hormones, neuropeptides, chemokines, odorants, phospholipids, and functional equivalents thereof that bind and preferably inhibit 184P1E2 protein function. Such non-limiting small molecules preferably have a molecular weight of less than about 10 kDa, more preferably below about 9, about 8, about 7, about 6, about 5 or about 4 kDa. In certain embodiments, small molecules physically associate with, or bind, 184P1E2 protein; are not found in naturally occurring metabolic pathways; and/or are more soluble in aqueous than non-aqueous solutions
“Stringency” of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured nucleic acid sequences to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature that can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).
“Stringent conditions” or “high stringency conditions”, as defined herein, are identified by, but not limited to, those that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.; (2) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42° C.; or (3) employ 50% formamide, 5×SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5×Denhardt's solution, sonicated salmon sperm DNA (50 μg/ml), 0.1% SDS, and 10% dextran sulfate at 42° C., with washes at 42° C. in 0.2×SSC (sodium chloride/sodium. citrate) and 50% formamide at 55° C., followed by a high-stringency wash consisting of 0.1×SSC containing EDTA at 55° C. “Moderately stringent conditions” are described by, but not limited to, those in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and include the use of washing solution and hybridization conditions (e.g., temperature, ionic strength and % SDS) less stringent than those described above. An example of moderately stringent conditions is overnight incubation at 37° C. in a solution comprising: 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 mg/mL denatured sheared salmon sperm DNA, followed by washing the filters in 1×SSC at about 37-50° C. The skilled artisan will recognize how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and the like.
An HLA “supermotif” is a peptide binding specificity shared by HLA molecules encoded by two or more HLA alleles.
As used herein “to treat” or “therapeutic” and grammatically related terms, refer to any improvement of any consequence of disease, such as prolonged survival, less morbidity, and/or a lessening of side effects which are the byproducts of an alternative therapeutic modality; full eradication of disease is not required.
A “transgenic animal” (e.g., a mouse or rat) is an animal having cells that contain a transgene, which transgene was introduced into the animal or an ancestor of the animal at a prenatal, e.g., an embryonic stage. A “transgene” is a DNA that is integrated into the genome of a cell from which a transgenic animal develops.
As used herein, an HLA or cellular immune response “vaccine” is a composition that contains or encodes one or more peptides of the invention. There are numerous embodiments of such vaccines, such as a cocktail of one or more individual peptides; one or more peptides of the invention comprised by a polyepitopic peptide; or nucleic acids that encode such individual peptides or polypeptides, e.g., a minigene that encodes a polyepitopic peptide. The “one or more peptides” can include any whole unit integer from 1-150 or more, e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150 or more peptides of the invention. The peptides or polypeptides can optionally be modified, such as by lipidation, addition of targeting or other sequences. HLA class I peptides of the invention can be admixed with, or linked to, HLA class II peptides, to facilitate activation of both cytotoxic T lymphocytes and helper T lymphocytes. HLA vaccines can also comprise peptide-pulsed antigen presenting cells, e.g., dendritic cells.
The term “variant” refers to a molecule that exhibits a variation from a described type or norm, such as a protein that has one or more different amino acid residues in the corresponding position(s) of a specifically described protein (e.g. the 184P1E2 protein shown in FIG. 2 or FIG. 3. An analog is an example of a variant protein. Splice isoforms and single nucleotides polymorphisms (SNPs) are further examples of variants.
The “184P1E2-related proteins” of the invention include those specifically identified herein, as well as allelic variants, conservative substitution variants, analogs and homologs that can be isolated/generated and characterized without undue experimentation following the methods outlined herein or readily available in the art. Fusion proteins that combine parts of different 184P1E2 proteins or fragments thereof, as well as fusion proteins of a 184P1E2 protein and a heterologous polypeptide are also included. Such 184P1E2 proteins are collectively referred to as the 184P1E2-related proteins, the proteins of the invention, or 184P1E2. The term “184P1E2-related protein” refers to a polypeptide fragment or a 184P1E2 protein sequence of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 amino acids; or, at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, or 664 or more amino acids.
II.) 184P1E2 Polynucleotides
One aspect of the invention provides polynucleotides corresponding or complementary to all or part of a 184P1E2 gene, mRNA, and/or coding sequence, preferably in isolated form, including polynucleotides encoding a 184P1E2-related protein and fragments thereof, DNA, RNA, DNA/RNA hybrid, and related molecules, polynucleotides or oligonucleotides complementary to a 184P1E2 gene or mRNA sequence or a part thereof, and polynucleotides or oligonucleotides that hybridize to a 184P1E2 gene, mRNA, or to a 184P1E2 encoding polynucleotide (collectively, “184P1E2 polynucleotides”). In all instances when referred to in this section, T can also be U in FIG. 2.
Embodiments of a 184P1E2 polynucleotide include: a 184P1E2 polynucleotide having the sequence shown in FIG. 2, the nucleotide sequence of 184P1E2 as shown in FIG. 2 wherein T is U; at least 10 contiguous nucleotides of a polynucleotide having the sequence as shown in FIG. 2; or, at least 10 contiguous nucleotides of a polynucleotide having the sequence as shown in FIG. 2 where T is U. For example, embodiments of 184P1E2 nucleotides comprise, without limitation:
(I) a polynucleotide comprising, consisting essentially of, or consisting of a sequence as shown in FIG. 2A (SEQ ID NO: 2), wherein T can also be U;
(II) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2A (SEQ ID NO: 2), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(III) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2B (SEQ ID NO: 4), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(IV) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2C (SEQ ID NO: 6), from nucleotide residue number 42 through nucleotide residue number 2036, including the a stop codon, wherein T can also be U;
(V) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2D (SEQ ID NO: 8), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(VI) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2E (SEQ ID NO: 10), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(VII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2F (SEQ ID NO: 12), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(VIII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2G (SEQ ID NO: 14), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(IX) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2H (SEQ ID NO: 16), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(X) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2I (SEQ ID NO: 18), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(XII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in FIG. 2J (SEQ ID NO: 20), from nucleotide residue number 42 through nucleotide residue number 2036, including the stop codon, wherein T can also be U;
(XIII) a polynucleotide that encodes a 184P1E2-related protein that is at least 90% homologous to an entire amino acid sequence shown in FIG. 2A-J (SEQ ID NO: 3);
(XIV) a polynucleotide that encodes a 184P1E2-related protein that is at least 90% identical to an entire amino acid sequence shown in FIG. 2A-J (SEQ ID NO: 5);
(XV) a polynucleotide that encodes at least one peptide set forth in Tables V-XVIII and XXII-LI;
(XVI) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of FIG. 3A in any whole number increment up to 664 that includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of FIG. 5;
(XVII) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of FIG. 3A in any whole number increment up to 664 that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of FIG. 6;
(XVIII) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of FIG. 3A in any whole number increment up to 664 that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of FIG. 7;
(XIX) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of FIG. 3A in any whole number increment up to 664 that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profile of FIG. 8;
(XX) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of FIG. 3A in any whole number increment up to 664 that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of FIG. 9;
(XXI) a polynucleotide that is fully complementary to a polynucleotide of any one of (I) —(XX).
(XXII) a peptide that is encoded by any of (I) —(XXI); and
(XXI) a polynucleotide of any of (I) —(XXI) or peptide of (XXII) together with a pharmaceutical excipient and/or in a human unit dose form.
As used herein, a range is understood to specifically disclose all whole unit positions thereof.
Typical embodiments of the invention disclosed herein include 184P1E2 polynucleotides that encode specific portions of 184P1E2 mRNA sequences (and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example:
(a) 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, or 664 or more contiguous amino acids of 184P1E2.
For example, representative embodiments of the invention disclosed herein include: polynucleotides and their encoded peptides themselves encoding about amino acid 1 to about amino acid 10 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 10 to about amino acid 20 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 20 to about amino acid 30 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 30 to about amino acid 40 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 40 to about amino acid 50 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 50 to about amino acid 60 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 60 to about amino acid 70 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 70 to about amino acid 80 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 80 to about amino acid 90 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, polynucleotides encoding about amino acid 90 to about amino acid 100 of the 184P1E2 protein shown in FIG. 2 or FIG. 3, in increments of about 10 amino acids, ending at the carboxyl terminal amino acid set forth in FIG. 2 or FIG. 3. Accordingly polynucleotides encoding portions of the amino acid sequence (of about 10 amino acids), of amino acids 100 through the carboxyl terminal amino acid of the 184P1E2 protein are embodiments of the invention. Wherein it is understood that each particular amino acid position discloses that position plus or minus five amino acid residues.
Polynucleotides encoding relatively long portions of a 184P1E2 protein are also within the scope of the invention. For example, polynucleotides encoding from about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 30, or 40 or 50 etc.) of the 184P1E2 protein “or variant” shown in FIG. 2 or FIG. 3 can be generated by a variety of techniques well known in the art. These polynucleotide fragments can include any portion of the 184P1E2 sequence as shown in FIG. 2.
Additional illustrative embodiments of the invention disclosed herein include 184P1E2 polynucleotide fragments encoding one or more of the biological motifs contained within a 184P1E2 protein “or variant” sequence, including one or more of the motif-bearing subsequences of a 184P1E2 protein “or variant” set forth in Tables V-XVIII and XXII-LI. In another embodiment, typical polynucleotide fragments of the invention encode one or more of the regions of 184P1E2 protein or variant that exhibit homology to a known molecule. In another embodiment of the invention, typical polynucleotide fragments can encode one or more of the 184P1E2 protein or variant N-glycosylation sites, cAMP and cGMP-dependent protein kinase phosphorylation sites, casein kinase II phosphorylation sites or N-myristoylation site and amidation sites.
II.A.) Uses of 184P1E2 Polynucleotides
II.A.I.) Monitoring of Genetic Abnormalities
The polynucleotides of the preceding paragraphs have a number of different specific uses. The human 184P1E2 gene maps to the chromosomal location set forth in the Example entitled “Chromosomal Mapping of 184P1E2.” For example, because the 184P1E2 gene maps to this chromosome, polynucleotides that encode different regions of the 184P1E2 proteins are used to characterize cytogenetic abnormalities of this chromosomal locale, such as abnormalities that are identified as being associated with various cancers. In certain genes, a variety of chromosomal abnormalities including rearrangements have been identified as frequent cytogenetic abnormalities in a number of different cancers (see e.g. Krajinovic et al., Mutat. Res. 382(3-4): 81-83 (1998); Johansson et al., Blood 86(10): 3905-3914 (1995) and Finger et al., P.N.A.S. 85(23): 9158-9162 (1988)). Thus, polynucleotides encoding specific regions of the 184P1E2 proteins provide new tools that can be used to delineate, with greater precision than previously possible, cytogenetic abnormalities in the chromosomal region that encodes 184P1E2 that may contribute to the malignant phenotype. In this context, these polynucleotides satisfy a need in the art for expanding the sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see e.g. Evans et al., Am. J. Obstet. Gynecol 171(4): 1055-1057 (1994)).
Furthermore, as 184P1E2 was shown to be highly expressed in bladder and other cancers, 184P1E2 polynucleotides are used in methods assessing the status of 184P1E2 gene products in normal versus cancerous tissues. Typically, polynucleotides that encode specific regions of the 184P1E2 proteins are used to assess the presence of perturbations (such as deletions, insertions, point mutations, or alterations resulting in a loss of an antigen etc.) in specific regions of the 184P1E2 gene, such as regions containing one or more motifs. Exemplary assays include both RT-PCR assays as well as single-strand conformation polymorphism (SSCP) analysis (see, e.g., Marrogi et al., J. Cutan. Pathol. 26(8): 369-378 (1999), both of which utilize polynucleotides encoding specific regions of a protein to examine these regions within the protein.
II.A.2.) Anti sense Embodiments
Other specifically contemplated nucleic acid related embodiments of the invention disclosed herein are genomic DNA, cDNAs, ribozymes, and antisense molecules, as well as nucleic acid molecules based on an alternative backbone, or including alternative bases, whether derived from natural sources or synthesized, and include molecules capable of inhibiting the RNA or protein expression of 184P1E2. For example, antisense molecules can be RNAs or other molecules, including peptide nucleic acids (PNAs) or non-nucleic acid molecules such as phosphorothioate derivatives, that specifically bind DNA or RNA in a base pair-dependent manner. A skilled artisan can readily obtain these classes of nucleic acid molecules using the 184P1E2 polynucleotides and polynucleotide sequences disclosed herein.
Antisense technology entails the administration of exogenous oligonucleotides that bind to a target polynucleotide located within the cells. The term “antisense” refers to the fact that such oligonucleotides are complementary to their intracellular targets, e.g., 184P1E2. See for example, Jack Cohen, Oligodeoxynucleotides, Antisense Inhibitors of Gene Expression, CRC Press, 1989; and Synthesis 1:1-5 (1988). The 184P1E2 antisense oligonucleotides of the present invention include derivatives such as S-oligonucleotides (phosphorothioate derivatives or S-oligos, see, Jack Cohen, supra), which exhibit enhanced cancer cell growth inhibitory action. S-oligos (nucleoside phosphorothioates) are isoelectronic analogs of an oligonucleotide (O-oligo) in which a nonbridging oxygen atom of the phosphate group is replaced by a sulfur atom. The S-oligos of the present invention can be prepared by treatment of the corresponding O-oligos with 3H-1,2-benzodithiol-3-one-1,1-dioxide, which is a sulfur transfer reagent. See, e.g., Iyer, R. P. et al., J. Org. Chem. 55:4693-4698 (1990); and Iyer, R. P. et al., J. Am. Chem. Soc. 112:1253-1254 (1990). Additional 184P1E2 antisense oligonucleotides of the present invention include morpholino antisense oligonucleotides known in the art (see, e.g., Partridge et al., 1996, Antisense & Nucleic Acid Drug Development 6: 169-175).
The 184P1E2 antisense oligonucleotides of the present invention typically can be RNA or DNA that is complementary to and stably hybridizes with the first 100 5′ codons or last 100 3′ codons of a 184P1E2 genomic sequence or the corresponding mRNA. Absolute complementarity is not required, although high degrees of complementarity are preferred. Use of an oligonucleotide complementary to this region allows for the selective hybridization to 184P1E2 mRNA and not to mRNA specifying other regulatory subunits of protein kinase. In one embodiment, 184P1E2 antisense oligonucleotides of the present invention are 15 to 30-mer fragments of the antisense DNA molecule that have a sequence that hybridizes to 184P1E2 mRNA. Optionally, 184P1E2 antisense oligonucleotide is a 30-mer oligonucleotide that is complementary to a region in the first 10 5′ codons or last 10 3′ codons of 184P1E2. Alternatively, the antisense molecules are modified to employ ribozymes in the inhibition of 184P1E2 expression, see, e.g., L. A. Couture & D. T. Stinchcomb; Trends Genet. 12: 510-515 (1996).
II.A.3.) Primers and Primer Pairs
Further specific embodiments of this nucleotides of the invention include primers and primer pairs, which allow the specific amplification of polynucleotides of the invention or of any specific parts thereof, and probes that selectively or specifically hybridize to nucleic acid molecules of the invention or to any part thereof. Probes can be labeled with a detectable marker, such as, for example, a radioisotope, fluorescent compound, bioluminescent compound, a chemiluminescent compound, metal chelator or enzyme. Such probes and primers are used to detect the presence of a 184P1E2 polynucleotide in a sample and as a means for detecting a cell expressing a 184P1E2 protein.
Examples of such probes include polypeptides comprising all or part of the human 184P1E2 cDNA sequence shown in FIG. 2. Examples of primer pairs capable of specifically amplifying 184P1E2 mRNAs are also described in the Examples. As will be understood by the skilled artisan, a great many different primers and probes can be prepared based on the sequences provided herein and used effectively to amplify and/or detect a 184P1E2 mRNA.
The 184P1E2 polynucleotides of the invention are useful for a variety of purposes, including but not limited to their use as probes and primers for the amplification and/or detection of the 184P1E2 gene(s), mRNA(s), or fragments thereof; as reagents for the diagnosis and/or prognosis of prostate cancer and other cancers; as coding sequences capable of directing the expression of 184P1E2 polypeptides; as tools for modulating or inhibiting the expression of the 184P1E2 gene(s) and/or translation of the 184P1E2 transcript(s); and as therapeutic agents.
The present invention includes the use of any probe as described herein to identify and isolate a 184P1E2 or 184P1E2 related nucleic acid sequence from a naturally occurring source, such as humans or other mammals, as well as the isolated nucleic acid sequence per se, which would comprise all or most of the sequences found in the probe used.
II.A.4.) Isolation of 184P1E2-Encoding Nucleic Acid Molecules
The 184P1E2 cDNA sequences described herein enable the isolation of other polynucleotides encoding 184P1E2 gene product(s), as well as the isolation of polynucleotides encoding 184P1E2 gene product homologs, alternatively spliced isoforms, allelic variants, and mutant forms of a 184P1E2 gene product as well as polynucleotides that encode analogs of 184P1E2-related proteins. Various molecular cloning methods that can be employed to isolate full length cDNAs encoding a 184P1E2 gene are well known (see, for example, Sambrook, J. et al., Molecular Cloning: A Laboratory Manual, 2d edition, Cold Spring Harbor Press, New York, 1989; Current Protocols in Molecular Biology. Ausubel et al., Eds., Wiley and Sons, 1995). For example, lambda phage cloning methodologies can be conveniently employed, using commercially available cloning systems (e.g., Lambda ZAP Express, Stratagene). Phage clones containing 184P1E2 gene cDNAs can be identified by probing with a labeled 184P1E2 cDNA or a fragment thereof. For example, in one embodiment, a 184P1E2 cDNA (e.g., FIG. 2) or a portion thereof can be synthesized and used as a probe to retrieve overlapping and full-length cDNAs corresponding to a 184P1E2 gene. A 184P1E2 gene itself can be isolated by screening genomic DNA libraries, bacterial artificial chromosome libraries (BACs), yeast artificial chromosome libraries (YACs), and the like, with 184P1E2 DNA probes or primers.
II.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems
The invention also provides recombinant DNA or RNA molecules containing a 184P1E2 polynucleotide, a fragment, analog or homologue thereof, including but not limited to phages, plasmids, phagemids, cosmids, YACs, BACs, as well as various viral and non-viral vectors well known in the art, and cells transformed or transfected with such recombinant DNA or RNA molecules. Methods for generating such molecules are well known (see, for example, Sambrook et al., 1989, supra).
The invention further provides a host-vector system comprising a recombinant DNA molecule containing a 184P1E2 polynucleotide, fragment, analog or homologue thereof within a suitable prokaryotic or eukaryotic host cell. Examples of suitable eukaryotic host cells include a yeast cell, a plant cell, or an animal cell, such as a mammalian cell or an insect cell (e.g., a baculovirus-infectible cell such as an Sf9 or HighFive cell). Examples of suitable mammalian cells include various prostate cancer cell lines such as DU145 and TsuPr1, other transfectable or transducible prostate cancer cell lines, primary cells (PrEC), as well as a number of mammalian cells routinely used for the expression of recombinant proteins (e.g., COS, CHO, 293, 293T cells). More particularly, a polynucleotide comprising the coding sequence of 184P1E2 or a fragment, analog or homolog thereof can be used to generate 184P1E2 proteins or fragments thereof using any number of host-vector systems routinely used and widely known in the art.
A wide range of host-vector systems suitable for the expression of 184P1E2 proteins or fragments thereof are available, see for example, Sambrook et al., 1989, supra; Current Protocols in Molecular Biology, 1995, supra). Preferred vectors for mammalian expression include but are not limited to pcDNA 3.1 myc-His-tag (Invitrogen) and the retroviral vector pSRαtkneo (Muller et al., 1991, MCB 11:1785). Using these expression vectors, 184P1E2 can be expressed in several prostate cancer and non-prostate cell lines, including for example 293, 293T, rat-1, NIH 3T3 and TsuPr1. The host-vector systems of the invention are useful for the production of a 184P1E2 protein or fragment thereof. Such host-vector systems can be employed to study the functional properties of 184P1E2 and 184P1E2 mutations or analogs.
Recombinant human 184P1E2 protein or an analog or homolog or fragment thereof can be produced by mammalian cells transfected with a construct encoding a 184P1E2-related nucleotide. For example, 293T cells can be transfected with an expression plasmid encoding 184P1E2 or fragment, analog or homolog thereof, a 184P1E2-related protein is expressed in the 293T cells, and the recombinant 184P1E2 protein is isolated using standard purification methods (e.g., affinity purification using anti-184P1E2 antibodies). In another embodiment, a 184P1E2 coding sequence is subcloned into the retroviral vector pSRαMSVtkneo and used to infect various mammalian cell lines, such as NIH 3T3, TsuPr1, 293 and rat-1 in order to establish 184P1E2 expressing cell lines. Various other expression systems well known in the art can also be employed. Expression constructs encoding a leader peptide joined in frame to a 184P1E2 coding sequence can be used for the generation of a secreted form of recombinant 184P1E2 protein.
As discussed herein, redundancy in the genetic code permits variation in 184P1E2 gene sequences. In particular, it is known in the art that specific host species often have specific codon preferences, and thus one can adapt the disclosed sequence as preferred for a desired host. For example, preferred analog codon sequences typically have rare codons (i.e., codons having a usage frequency of less than about 20% in known sequences of the desired host) replaced with higher frequency codons. Codon preferences for a specific species are calculated, for example, by utilizing codon usage tables available on the INTERNET.
Additional sequence modifications are known to enhance protein expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon/intron splice site signals, transposon-like repeats, and/or other such well-characterized sequences that are deleterious to gene expression. The GC content of the sequence is adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell. Where possible, the sequence is modified to avoid predicted hairpin secondary mRNA structures. Other useful modifications include the addition of a translational initiation consensus sequence at the start of the open reading frame, as described in Kozak, Mol. Cell Biol., 9:5073-5080 (1989). Skilled artisans understand that the general rule that eukaryotic ribosomes initiate translation exclusively at the 5′ proximal AUG codon is abrogated only under rare conditions (see, e.g., Kozak PNAS 92(7): 2662-2666, (1995) and Kozak NAR 15(20): 8125-8148 (1987)).
III.) 184P1E2-Related Proteins
Another aspect of the present invention provides 184P1E2-related proteins. Specific embodiments of 184P1E2 proteins comprise a polypeptide having all or part of the amino acid sequence of human 184P1E2 as shown in FIG. 2 or FIG. 3. Alternatively, embodiments of 184P1E2 proteins comprise variant, homolog or analog polypeptides that have alterations in the amino acid sequence of 184P1E2 shown in FIG. 2 or FIG. 3.
In general, naturally occurring allelic variants of human 184P1E2 share a high degree of structural identity and homology (e.g., 90% or more homology). Typically, allelic variants of a 184P1E2 protein contain conservative amino acid substitutions within the 184P1E2 sequences described herein or contain a substitution of an amino acid from a corresponding position in a homologue of 184P1E2. One class of 184P1E2 allelic variants are proteins that share a high degree of homology with at least a small region of a particular 184P1E2 amino acid sequence, but further contain a radical departure from the sequence, such as a non-conservative substitution, truncation, insertion or frame shift. In comparisons of protein sequences, the terms, similarity, identity, and homology each have a distinct meaning as appreciated in the field of genetics. Moreover, orthology and paralogy can be important concepts describing the relationship of members of a given protein family in one organism to the members of the same family in other organisms.
Amino acid abbreviations are provided in Table H. Conservative amino acid substitutions can frequently be made in a protein without altering either the conformation or the function of the protein. Proteins of the invention can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 conservative substitutions. Such changes include substituting any of isoleucine (I), valine (V), and leucine (L) for any other of these hydrophobic amino acids; aspartic acid (D) for glutamic acid (E) and vice versa; glutamine (Q) for asparagine (N) and vice versa; and serine (S) for threonine (T) and vice versa. Other substitutions can also be considered conservative, depending on the environment of the particular amino acid and its role in the three-dimensional structure of the protein. For example, glycine (G) and alanine (A) can frequently be interchangeable, as can alanine (A) and valine (V). Methionine (M), which is relatively hydrophobic, can frequently be interchanged with leucine and isoleucine, and sometimes with valine. Lysine (K) and arginine (R) are frequently interchangeable in locations in which the significant feature of the amino acid residue is its charge and the differing pK's of these two amino acid residues are not significant. Still other changes can be considered “conservative” in particular environments (see, e.g. Table III herein; pages 13-15 “Biochemistry” 2nd ED. Lubert Stryer ed (Stanford University); Henikoff et al., PNAS 1992 Vol 89 10915-10919; Lei et al., J Biol Chem 1995 May 19; 270(20):11882-6).
Embodiments of the invention disclosed herein include a wide variety of art-accepted variants or analogs of 184P1E2 proteins such as polypeptides having amino acid insertions, deletions and substitutions. 184P1E2 variants can be made using methods known in the art such as site-directed mutagenesis, alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (Carter et al., Nucl. Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res., 10:6487 (1987)), cassette mutagenesis (Wells et al., Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et al., Philos. Trans. R. Soc. London SerA, 317:415 (1986)) or other known techniques can be performed on the cloned DNA to produce the 184P1E2 variant DNA.
Scanning amino acid analysis can also be employed to identify one or more amino acids along a contiguous sequence that is involved in a specific biological activity such as a protein-protein interaction. Among the preferred scanning amino acids are relatively small, neutral amino acids. Such amino acids include alanine, glycine, serine, and cysteine. Alanine is typically a preferred scanning amino acid among this group because it eliminates the side-chain beyond the beta-carbon and is less likely to alter the main-chain conformation of the variant. Alanine is also typically preferred because it is the most common amino acid. Further, it is frequently found in both buried and exposed positions (Creighton, The Proteins, (W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150:1 (1976)). If alanine substitution does not yield adequate amounts of variant, an isosteric amino acid can be used.
As defined herein, 184P1E2 variants, analogs or homologs, have the distinguishing attribute of having at least one epitope that is “cross reactive” with a 184P1E2 protein having an amino acid sequence of FIG. 3. As used in this sentence, “cross reactive” means that an antibody or T cell that specifically binds to a 184P1E2 variant also specifically binds to a 184P1E2 protein having an amino acid sequence set forth in FIG. 3. A polypeptide ceases to be a variant of a protein shown in FIG. 3, when it no longer contains any epitope capable of being recognized by an antibody or T cell that specifically binds to the starting 184P1E2 protein. Those skilled in the art understand that antibodies that recognize proteins bind to epitopes of varying size, and a grouping of the order of about four or five amino acids, contiguous or not, is regarded as a typical number of amino acids in a minimal epitope. See, e.g., Nair et al., J. Immunol 2000 165(12): 6949-6955; Hebbes et al., Mol Immunol (1989) 26(9):865-73; Schwartz et al., J Immunol (1985) 135(4):2598-608.
Other classes of 184P1E2-related protein variants share 70%, 75%, 80%, 85% or 90% or more similarity with an amino acid sequence of FIG. 3, or a fragment thereof. Another specific class of 184P1E2 protein variants or analogs comprise one or more of the 184P1E2 biological motifs described herein or presently known in the art. Thus, encompassed by the present invention are analogs of 184P1E2 fragments (nucleic or amino acid) that have altered functional (e.g. immunogenic) properties relative to the starting fragment. It is to be appreciated that motifs now or which become part of the art are to be applied to the nucleic or amino acid sequences of FIG. 2 or 3.
As discussed herein, embodiments of the claimed invention include polypeptides containing less than the full amino acid sequence of a 184P1E2 protein shown in FIG. 2 or FIG. 3. For example, representative embodiments of the invention comprise peptides/proteins having any 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino acids of a 184P1E2 protein shown in FIG. 2 or FIG. 3.
Moreover, representative embodiments of the invention disclosed herein include polypeptides consisting of about amino acid 1 to about amino acid 10 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 10 to about amino acid 20 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 20 to about amino acid 30 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 30 to about amino acid 40 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 40 to about amino acid 50 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 50 to about amino acid 60 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 60 to about amino acid 70 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 70 to about amino acid 80 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 80 to about amino acid 90 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, polypeptides consisting of about amino acid 90 to about amino acid 100 of a 184P1E2 protein shown in FIG. 2 or FIG. 3, etc. throughout the entirety of a 184P1E2 amino acid sequence. Moreover, polypeptides consisting of about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 130, or 140 or 150 etc.) of a 184P1E2 protein shown in FIG. 2 or FIG. 3 are embodiments of the invention. It is to be appreciated that the starting and stopping positions in this paragraph refer to the specified position as well as that position plus or minus 5 residues.
184P1E2-related proteins are generated using standard peptide synthesis technology or using chemical cleavage methods well known in the art. Alternatively, recombinant methods can be used to generate nucleic acid molecules that encode a 184P1E2-related protein. In one embodiment, nucleic acid molecules provide a means to generate defined fragments of a 184P1E2 protein (or variants, homologs or analogs thereof).
III.A.) Motif-Bearing Protein Embodiments
Additional illustrative embodiments of the invention disclosed herein include 184P1E2 polypeptides comprising the amino acid residues of one or more of the biological motifs contained within a 184P1E2 polypeptide sequence set forth in FIG. 2 or FIG. 3. Various motifs are known in the art, and a protein can be evaluated for the presence of such motifs by a number of publicly available Internet sites (see, e.g., EpimatrixTM and EpimerTM, Brown University, and BIMAS.
Motif bearing subsequences of all 184P1E2 variant proteins are set forth and identified in Tables V-XVIII and XXII-LI.
Table XIX sets forth several frequently occurring motifs based on pfam searches (see URL address pfam.wustl.edu/). The columns of Table XIX list (1) motif name abbreviation, (2) percent identity found amongst the different member of the motif family, (3) motif name or description and (4) most common function; location information is included if the motif is relevant for location.
Polypeptides comprising one or more of the 184P1E2 motifs discussed above are useful in elucidating the specific characteristics of a malignant phenotype in view of the observation that the 184P1E2 motifs discussed above are associated with growth dysregulation and because 184P1E2 is overexpressed in certain cancers (See, e.g., Table I). Casein kinase II, cAMP and camp-dependent protein kinase, and Protein Kinase C, for example, are enzymes known to be associated with the development of the malignant phenotype (see e.g. Chen et al., Lab Invest., 78(2): 165-174 (1998); Gaiddon et al., Endocrinology 136(10): 4331-4338 (1995); Hall et al., Nucleic Acids Res. 24(6): 1119-1126 (1996); Peterziel et al., Oncogene 18(46): 6322-6329 (1999) and O'Brian, Oncol. Rep. 5(2): 305-309 (1998)). Moreover, both glycosylation and myristoylation are protein modifications also associated with cancer and cancer progression (see e.g. Dennis et al., Biochem. Biophys. Acta 1473(1):21-34 (1999); Raju et al., Exp. Cell Res. 235(1): 145-154 (1997)). Amidation is another protein modification also associated with cancer and cancer progression (see e.g. Treston et al., J. Natl. Cancer Inst. Monogr. (13): 169-175 (1992)).
In another embodiment, proteins of the invention comprise one or more of the immunoreactive epitopes identified in accordance with art-accepted methods, such as the peptides set forth in Tables V-XVIII and XXII-LI. CTL epitopes can be determined using specific algorithms to identify peptides within a 184P1E2 protein that are capable of optimally binding to specified HLA alleles (e.g., Table IV; EpimatrixTM and EpimerTM, Brown University, and BIMAS). Moreover, processes for identifying peptides that have sufficient binding affinity for HLA molecules and which are correlated with being immunogenic epitopes, are well known in the art, and are carried out without undue experimentation. In addition, processes for identifying peptides that are immunogenic epitopes, are well known in the art, and are carried out without undue experimentation either in vitro or in vivo.
Also known in the art are principles for creating analogs of such epitopes in order to modulate immunogenicity. For example, one begins with an epitope that bears a CTL or HTL motif (see, e.g., the HLA Class I and HLA Class II motifs/supermotifs of Table IV). The epitope is analoged by substituting out an amino acid at one of the specified positions, and replacing it with another amino acid specified for that position. For example, one can substitute out a deleterious residue in favor of any other residue, such as a preferred residue as defined in Table IV; substitute a less-preferred residue with a preferred residue as defined in Table IV; or substitute an originally-occurring preferred residue with another preferred residue as defined in Table IV. Substitutions can occur at primary anchor positions or at other positions in a peptide; see, e.g., Table IV.
A variety of references reflect the art regarding the identification and generation of epitopes in a protein of interest as well as analogs thereof. See, for example, WO 97/33602 to Chesnut et al.; Sette, Immunogenetics 1999 50(3-4): 201-212; Sette et al., J. Immunol. 2001 166(2): 1389-1397; Sidney et al., Hum. Immunol. 1997 58(1): 12-20; Kondo et al., Immunogenetics 1997 45(4): 249-258; Sidney et al., J. Immunol. 1996 157(8): 3480-90; and Falk et al., Nature 351: 290-6 (1991); Hunt et al., Science 255:1261-3 (1992); Parker et al., J. Immunol. 149:3580-7 (1992); Parker et al., J. Immunol. 152:163-75 (1994)); Kast et al., 1994 152(8): 3904-12; Borras-Cuesta et al., Hum. Immunol. 2000 61(3): 266-278; Alexander et al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., PMID: 7895164, UI: 95202582; O'Sullivan et al., J. Immunol. 1991 147(8): 2663-2669; Alexander et al., Immunity 1994 1(9): 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92.
Related embodiments of the invention include polypeptides comprising combinations of the different motifs set forth in Table XX, and/or, one or more of the predicted CTL epitopes of Tables V-XVII and XXII-XLVII, and/or, one or more of the predicted HTL epitopes of Tables XLVIII-LI, and/or, one or more of the T cell binding motifs known in the art. Preferred embodiments contain no insertions, deletions or substitutions either within the motifs or the intervening sequences of the polypeptides. In addition, embodiments which include a number of either N-terminal and/or C-terminal amino acid residues on either side of these motifs may be desirable (to, for example, include a greater portion of the polypeptide architecture in which the motif is located). Typically the number of N-terminal and/or C-terminal amino acid residues on either side of a motif is between about 1 to about 100 amino acid residues, preferably 5 to about 50 amino acid residues.
184P1E2-related proteins are embodied in many forms, preferably in isolated form. A purified 184P1E2 protein molecule will be substantially free of other proteins or molecules that impair the binding of 184P1E2 to antibody, T cell or other ligand. The nature and degree of isolation and purification will depend on the intended use. Embodiments of a 184P1E2-related proteins include purified 184P1E2-related proteins and functional, soluble 184P1E2-related proteins. In one embodiment, a functional, soluble 184P1E2 protein or fragment thereof retains the ability to be bound by antibody, T cell or other ligand.
The invention also provides 184P1E2 proteins comprising biologically active fragments of a 184P1E2 amino acid sequence shown in FIG. 2 or FIG. 3. Such proteins exhibit properties of the starting 184P1E2 protein, such as the ability to elicit the generation of antibodies that specifically bind an epitope associated with the starting 184P1E2 protein; to be bound by such antibodies; to elicit the activation of HTL or CTL; and/or, to be recognized by HTL or CTL that also specifically bind to the starting protein.
184P1E2-related polypeptides that contain particularly interesting structures can be predicted and/or identified using various analytical techniques well known in the art, including, for example, the methods of Chou-Fasman, Garnier-Robson, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis, or on the basis of immunogenicity. Fragments that contain such structures are particularly useful in generating subunit-specific anti-184P1E2 antibodies, or T cells or in identifying cellular factors that bind to 184P1E2. For example, hydrophilicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Hopp, T. P. and Woods, K. R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 157:105-132. Percent (%) Accessible Residues profiles can be generated, and immunogenic peptide fragments identified, using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated, and immunogenic peptide fragments identified, using the method of Bhaskaran R., Ponnuswamy P. K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated, and immunogenic peptide fragments identified, using the method of Deleage, G., Roux B., 1987, Protein Engineering 1:289-294.
CTL epitopes can be determined using specific algorithms to identify peptides within a 184P1E2 protein that are capable of optimally binding to specified HLA alleles (e.g., by using the SYFPEITHI site on the INTERNET (the listings in Table IV(A)-(E)), EpimatrixTM and EpimerTM, Brown University, and BIMAS). Illustrating this, peptide epitopes from 184P1E2 that are presented in the context of human MHC Class I molecules, e.g., HLA-A1, A2, A3, A11, A24, B7 and B35 were predicted (see, e.g., Tables V-XVIII, XXII-LI). Specifically, the complete amino acid sequence of the 184P1E2 protein and relevant portions of other variants, i.e., for HLA Class I predictions 9 flanking residues on either side of a point mutation, and for HLA Class II predictions 14 flanking residues on either side of a point mutation, were entered into the HLA Peptide Motif Search algorithm found in the Bioinformatics and Molecular Analysis Section (BIMAS) web site listed above; in addition to the INTERNET site SYFPEITHI.
The HLA peptide motif search algorithm was developed by Dr. Ken Parker based on binding of specific peptide sequences in the groove of HLA Class I molecules, in particular HLA-A2 (see, e.g., Falk et al., Nature 351: 290-6 (1991); Hunt et al., Science 255:1261-3 (1992); Parker et al., J. Immunol. 149:3580-7 (1992); Parker et al., J. Immunol. 152:163-75 (1994)). This algorithm allows location and ranking of 8-mer, 9-mer, and 10-mer peptides from a complete protein sequence for predicted binding to HLA-A2 as well as numerous other HLA Class I molecules. Many HLA class I binding peptides are 8-, 9-, 10 or 11-mers. For example, for Class I HLA-A2, the epitopes preferably contain a leucine (L) or methionine (M) at position 2 and a valine (V) or leucine (L) at the C-terminus (see, e.g., Parker et al., J. Immunol. 149:3580-7 (1992)). Selected results of 184P1E2 predicted binding peptides are shown in Tables V-XVIII and XXII-LI herein. In Tables V-XVIII and XXII-XLVII, selected candidates, 9-mers and 10-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. In Tables XLVIII-LI, selected candidates, 15-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. The binding score corresponds to the estimated half time of dissociation of complexes containing the peptide at 37° C. at pH 6.5. Peptides with the highest binding score are predicted to be the most tightly bound to HLA Class I on the cell surface for the greatest period of time and thus represent the best immunogenic targets for T-cell recognition.
Actual binding of peptides to an HLA allele can be evaluated by stabilization of HLA expression on the antigen-processing defective cell line T2 (see, e.g., Xue et al., Prostate 30:73-8 (1997) and Peshwa et al., Prostate 36:129-38 (1998)). Immunogenicity of specific peptides can be evaluated in vitro by stimulation of CD8+ cytotoxic T lymphocytes (CTL) in the presence of antigen presenting cells such as dendritic cells.
It is to be appreciated that every epitope predicted by the BIMAS site, EpimerTM and EpimatrixTM sites, or specified by the HLA class I or class II motifs available in the art or which become part of the art such as set forth in Table IV (or determined using World Wide Web site URL syfpeithi.bmi-heidelberg.com/, or BIMAS, bimas.dcrt.nih.gov/) are to be “applied” to a 184P1E2 protein in accordance with the invention. As used in this context “applied” means that a 184P1E2 protein is evaluated, e.g., visually or by computer-based patterns finding methods, as appreciated by those of skill in the relevant art. Every subsequence of a 184P1E2 protein of 8, 9, 10, or 11 amino acid residues that bears an HLA Class I motif, or a subsequence of 9 or more amino acid residues that bear an HLA Class II motif are within the scope of the invention.
III.B.) Expression of 184P1E2-Related Proteins
In an embodiment described in the examples that follow, 184P1E2 can be conveniently expressed in cells (such as 293T cells) transfected with a commercially available expression vector such as a CMV-driven expression vector encoding 184P1E2 with a C-terminal 6×His and MYC tag (pcDNA3.1/mycHIS, Invitrogen or Tag5, GenHunter Corporation, Nashville Tenn.). The Tag5 vector provides an IgGK secretion signal that can be used to facilitate the production of a secreted 184P1E2 protein in transfected cells. The secreted HIS-tagged 184P1E2 in the culture media can be purified, e.g., using a nickel column using standard techniques.
III.C.) Modifications of 184P1E2-Related Proteins
Modifications of 184P1E2-related proteins such as covalent modifications are included within the scope of this invention. One type of covalent modification includes reacting targeted amino acid residues of a 184P1E2 polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C-terminal residues of a 184P1E2 protein. Another type of covalent modification of a 184P1E2 polypeptide included within the scope of this invention comprises altering the native glycosylation pattern of a protein of the invention. Another type of covalent modification of 184P1E2 comprises linking a 184P1E2 polypeptide to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol (PEG), polypropylene glycol, or polyoxyalkylenes, in the manner set forth in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.
The 184P1E2-related proteins of the present invention can also be modified to form a chimeric molecule comprising 184P1E2 fused to another, heterologous polypeptide or amino acid sequence. Such a chimeric molecule can be synthesized chemically or recombinantly. A chimeric molecule can have a protein of the invention fused to another tumor-associated antigen or fragment thereof. Alternatively, a protein in accordance with the invention can comprise a fusion of fragments of a 184P1E2 sequence (amino or nucleic acid) such that a molecule is created that is not, through its length, directly homologous to the amino or nucleic acid sequences shown in FIG. 2 or FIG. 3. Such a chimeric molecule can comprise multiples of the same subsequence of 184P1E2. A chimeric molecule can comprise a fusion of a 184P1E2-related protein with a polyhistidine epitope tag, which provides an epitope to which immobilized nickel can selectively bind, with cytokines or with growth factors. The epitope tag is generally placed at the amino- or carboxyl-terminus of a 184P1E2 protein. In an alternative embodiment, the chimeric molecule can comprise a fusion of a 184P1E2-related protein with an immunoglobulin or a particular region of an immunoglobulin. For a bivalent form of the chimeric molecule (also referred to as an “immunoadhesin”), such a fusion could be to the Fc region of an IgG molecule. The Ig fusions preferably include the substitution of a soluble (transmembrane domain deleted or inactivated) form of a 184P1E2 polypeptide in place of at least one variable region within an Ig molecule. In a preferred embodiment, the immunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge, CH1, CH2 and CH3 regions of an IgGI molecule. For the production of immunoglobulin fusions see, e.g., U.S. Pat. No. 5,428,130 issued Jun. 27, 1995.
III.D.) Uses of 184P1E2-Related Proteins
The proteins of the invention have a number of different specific uses. As 184P1E2 is highly expressed in prostate and other cancers, 184P1E2-related proteins are used in methods that assess the status of 184P1E2 gene products in normal versus cancerous tissues, thereby elucidating the malignant phenotype. Typically, polypeptides from specific regions of a 184P1E2 protein are used to assess the presence of perturbations (such as deletions, insertions, point mutations etc.) in those regions (such as regions containing one or more motifs). Exemplary assays utilize antibodies or T cells targeting 184P1E2-related proteins comprising the amino acid residues of one or more of the biological motifs contained within a 184P1E2 polypeptide sequence in order to evaluate the characteristics of this region in normal versus cancerous tissues or to elicit an immune response to the epitope. Alternatively, 184P1E2-related proteins that contain the amino acid residues of one or more of the biological motifs in a 184P1E2 protein are used to screen for factors that interact with that region of 184P1E2.
184P1E2 protein fragments/subsequences are particularly useful in generating and characterizing domain-specific antibodies (e.g., antibodies recognizing an extracellular or intracellular epitope of a 184P1E2 protein), for identifying agents or cellular factors that bind to 184P1E2 or a particular structural domain thereof, and in various therapeutic and diagnostic contexts, including but not limited to diagnostic assays, cancer vaccines and methods of preparing such vaccines.
Proteins encoded by the 184P1E2 genes, or by analogs, homologs or fragments thereof, have a variety of uses, including but not limited to generating antibodies and in methods for identifying ligands and other agents and cellular constituents that bind to a 184P1E2 gene product. Antibodies raised against a 184P1E2 protein or fragment thereof are useful in diagnostic and prognostic assays, and imaging methodologies in the management of human cancers characterized by expression of 184P1E2 protein, such as those listed in Table I. Such antibodies can be expressed intracellularly and used in methods of treating patients with such cancers. 184P1E2-related nucleic acids or proteins are also used in generating HTL or CTL responses.
Various immunological assays useful for the detection of 184P1E2 proteins are used, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), immunocytochemical methods, and the like. Antibodies can be labeled and used as immunological imaging reagents capable of detecting 184P1E2-expressing cells (e.g., in radioscintigraphic imaging methods). 184P1E2 proteins are also particularly useful in generating cancer vaccines, as further described herein.
IV.) 184P1E2 Antibodies
Another aspect of the invention provides antibodies that bind to 184P1E2-related proteins. Preferred antibodies specifically bind to a 184P1E2-related protein and do not bind (or bind weakly) to peptides or proteins that are not 184P1E2-related proteins. For example, antibodies that bind 184P1E2 can bind 184P1E2-related proteins such as the homologs or analogs thereof.
184P1E2 antibodies of the invention are particularly useful in cancer (see, e.g., Table I) diagnostic and prognostic assays, and imaging methodologies. Similarly, such antibodies are useful in the treatment, diagnosis, and/or prognosis of other cancers, to the extent 184P1E2 is also expressed or overexpressed in these other cancers. Moreover, intracellularly expressed antibodies (e.g., single chain antibodies) are therapeutically useful in treating cancers in which the expression of 184P1E2 is involved, such as advanced or metastatic prostate cancers.
The invention also provides various immunological assays useful for the detection and quantification of 184P1E2 and mutant 184P1E2-related proteins. Such assays can comprise one or more 184P1E2 antibodies capable of recognizing and binding a 184P1E2-related protein, as appropriate. These assays are performed within various immunological assay formats well known in the art, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), and the like.
Immunological non-antibody assays of the invention also comprise T cell immunogenicity assays (inhibitory or stimulatory) as well as major histocompatibility complex (MHC) binding assays.
In addition, immunological imaging methods capable of detecting prostate cancer and other cancers expressing 184P1E2 are also provided by the invention, including but not limited to radioscintigraphic imaging methods using labeled 184P1E2 antibodies. Such assays are clinically useful in the detection, monitoring, and prognosis of 184P1E2 expressing cancers such as prostate cancer.
184P1E2 antibodies are also used in methods for purifying a 184P1E2-related protein and for isolating 184P1E2 homologues and related molecules. For example, a method of purifying a 184P1E2-related protein comprises incubating a 184P1E2 antibody, which has been coupled to a solid matrix, with a lysate or other solution containing a 184P1E2-related protein under conditions that permit the 184P1E2 antibody to bind to the 184P1E2-related protein; washing the solid matrix to eliminate impurities; and eluting the 184P1E2-related protein from the coupled antibody. Other uses of 184P1E2 antibodies in accordance with the invention include generating anti-idiotypic antibodies that mimic a 184P1E2 protein.
Various methods for the preparation of antibodies are well known in the art. For example, antibodies can be prepared by immunizing a suitable mammalian host using a 184P1E2-related protein, peptide, or fragment, in isolated or immunoconjugated form (Antibodies: A Laboratory Manual, CSH Press, Eds., Harlow, and Lane (1988); Harlow, Antibodies, Cold Spring Harbor Press, NY (1989)). In addition, fusion proteins of 184P1E2 can also be used, such as a 184P1E2 GST-fusion protein. In a particular embodiment, a GST fusion protein comprising all or most of the amino acid sequence of FIG. 2 or FIG. 3 is produced, then used as an immunogen to generate appropriate antibodies. In another embodiment, a 184P1E2-related protein is synthesized and used as an immunogen.
In addition, naked DNA immunization techniques known in the art are used (with or without purified 184P1E2-related protein or 184P1E2 expressing cells) to generate an immune response to the encoded immunogen (for review, see Donnelly et al., 1997, Ann. Rev. Immunol. 15: 617-648).
The amino acid sequence of a 184P1E2 protein as shown in FIG. 2 or FIG. 3 can be analyzed to select specific regions of the 184P1E2 protein for generating antibodies. For example, hydrophobicity and hydrophilicity analyses of a 184P1E2 amino acid sequence are used to identify hydrophilic regions in the 184P1E2 structure. Regions of a 184P1E2 protein that show immunogenic structure, as well as other regions and domains, can readily be identified using various other methods known in the art, such as Chou-Fasman, Garnier-Robson, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis. Hydrophilicity profiles can be generated using the method of Hopp, T. P. and Woods, K. R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated using the method of Kyte, J. and Doolittle, R. F., 1982, J. Mol. Biol. 157:105-132. Percent (%) Accessible Residues profiles can be generated using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated using the method of Bhaskaran R., Ponnuswamy P. K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated using the method of Deleage, G., Roux B., 1987, Protein Engineering 1:289-294. Thus, each region identified by any of these programs or methods is within the scope of the present invention. Methods for the generation of 184P1E2 antibodies are further illustrated by way of the examples provided herein. Methods for preparing a protein or polypeptide for use as an immunogen are well known in the art. Also well known in the art are methods for preparing immunogenic conjugates of a protein with a carrier, such as BSA, KLH or other carrier protein. In some circumstances, direct conjugation using, for example, carbodiimide reagents are used; in other instances linking reagents such as those supplied by Pierce Chemical Co., Rockford, Ill., are effective. Administration of a 184P1E2 immunogen is often conducted by injection over a suitable time period and with use of a suitable adjuvant, as is understood in the art. During the immunization schedule, titers of antibodies can be taken to determine adequacy of antibody formation.
184P1E2 monoclonal antibodies can be produced by various means well known in the art. For example, immortalized cell lines that secrete a desired monoclonal antibody are prepared using the standard hybridoma technology of Kohler and Milstein or modifications that immortalize antibody-producing B cells, as is generally known. Immortalized cell lines that secrete the desired antibodies are screened by immunoassay in which the antigen is a 184P1E2-related protein. When the appropriate immortalized cell culture is identified, the cells can be expanded and antibodies produced either from in vitro cultures or from ascites fluid.
The antibodies or fragments of the invention can also be produced, by recombinant means. Regions that bind specifically to the desired regions of a 184P1E2 protein can also be produced in the context of chimeric or complementarity determining region (CDR) grafted antibodies of multiple species origin. Humanized or human 184P1E2 antibodies can also be produced, and are preferred for use in therapeutic contexts. Methods for humanizing murine and other non-human antibodies, by substituting one or more of the non-human antibody CDRs for corresponding human antibody sequences, are well known (see for example, Jones et al., 1986, Nature 321: 522-525; Riechmann et al., 1988, Nature 332: 323-327; Verhoeyen et al., 1988, Science 239: 1534-1536). See also, Carter et al., 1993, Proc. Natl. Acad. Sci. USA 89: 4285 and Sims et al., 1993, J. Immunol. 151: 2296.
Methods for producing fully human monoclonal antibodies include phage display and transgenic methods (for review, see Vaughan et al., 1998, Nature Biotechnology 16: 535-539). Fully human 184P1E2 monoclonal antibodies can be generated using cloning technologies employing large human Ig gene combinatorial libraries (i.e., phage display) (Griffiths and Hoogenboom, Building an in vitro immune system: human antibodies from phage display libraries. In: Protein Engineering of Antibody Molecules for Prophylactic and Therapeutic Applications in Man, Clark, M. (Ed.), Nottingham Academic, pp 45-64 (1993); Burton and Barbas, Human Antibodies from combinatorial libraries. Id., pp 65-82). Fully human 184P1E2 monoclonal antibodies can also be produced using transgenic mice engineered to contain human immunoglobulin gene loci as described in PCT Patent Application WO98/24893, Kucherlapati and Jakobovits et al., published Dec. 3, 1997 (see also, Jakobovits, 1998, Exp. Opin. Invest. Drugs 7(4): 607-614; U.S. Pat. No. 6,162,963 issued 19 Dec. 2000; U.S. Pat. No. 6,150,584 issued 12 Nov. 2000; and, U.S. Pat. No. 6,114,598 issued 5 Sep. 2000). This method avoids the in vitro manipulation required with phage display technology and efficiently produces high affinity authentic human antibodies.
Reactivity of 184P1E2 antibodies with a 184P1E2-related protein can be established by a number of well known means, including Western blot, immunoprecipitation, ELISA, and FACS analyses using, as appropriate, 184P1E2-related proteins, 184P1E2-expressing cells or extracts thereof. A 184P1E2 antibody or fragment thereof can be labeled with a detectable marker or conjugated to a second molecule. Suitable detectable markers include, but are not limited to, a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme. Further, bi-specific antibodies specific for two or more 184P1E2 epitopes are generated using methods generally known in the art. Homodimeric antibodies can also be generated by cross-linking techniques known in the art (e.g., Wolff et al., Cancer Res. 53: 2560-2565).
V.) 184P1E2 Cellular Immune Responses
The mechanism by which T cells recognize antigens has been delineated. Efficacious peptide epitope vaccine compositions of the invention induce a therapeutic or prophylactic immune responses in very broad segments of the world-wide population. For an understanding of the value and efficacy of compositions of the invention that induce cellular immune responses, a brief review of immunology-related technology is provided.
A complex of an HLA molecule and a peptidic antigen acts as the ligand recognized by HLA-restricted T cells (Buus, S. et al., Cell 47:1071, 1986; Babbitt, B. P. et al., Nature 317:359, 1985; Townsend, A. and Bodmer, H., Annu. Rev. Immunol. 7:601, 1989; Germain, R. N., Annu. Rev. Immunol. 11:403, 1993). Through the study of single amino acid substituted antigen analogs and the sequencing of endogenously bound, naturally processed peptides, critical residues that correspond to motifs required for specific binding to HLA antigen molecules have been identified and are set forth in Table IV (see also, e.g., Southwood, et al., J. Immunol. 160:3363, 1998; Rammensee, et al., Immunogenetics 41:178, 1995; Rammensee et al., SYFPEITHI, access via World Wide Web at URL syfpeithi.bmi-heidelberg.com/; Sette, A. and Sidney, J. Curr. Opin. Immunol. 10:478, 1998; Engelhard, V. H., Curr. Opin. Immunol. 6:13, 1994; Sette, A. and Grey, H. M., Curr. Opin. Immunol. 4:79, 1992; Sinigaglia, F. and Hammer, J. Curr. Biol. 6:52, 1994; Ruppert et al., Cell 74:929-937, 1993; Kondo et al., J. Immunol. 155:4307-4312, 1995; Sidney et al., J. Immunol. 157:3480-3490, 1996; Sidney et al., Human Immunol. 45:79-93, 1996; Sette, A. and Sidney, J. Immunogenetics 1999 November; 50(3-4):201-12, Review).
Furthermore, x-ray crystallographic analyses of HLA-peptide complexes have revealed pockets within the peptide binding cleft/groove of HLA molecules which accommodate, in an allele-specific mode, residues borne by peptide ligands; these residues in turn determine the HLA binding capacity of the peptides in which they are present. (See, e.g., Madden, D. R. Annu. Rev. Immunol. 13:587, 1995; Smith, et al., Immunity 4:203, 1996; Fremont et al., Immunity 8:305, 1998; Stern et al., Structure 2:245, 1994; Jones, E. Y. Curr. Opin. Immunol. 9:75, 1997; Brown, J. H. et al., Nature 364:33, 1993; Guo, H. C. et al., Proc. Natl. Acad. Sci. USA 90:8053, 1993; Guo, H. C. et al., Nature 360:364, 1992; Silver, M. L. et al., Nature 360:367, 1992; Matsumura, M. et al., Science 257:927, 1992; Madden et al., Cell 70:1035, 1992; Fremont, D. H. et al., Science 257:919, 1992; Saper, M. A., Bjorkman, P. J. and Wiley, D.C., J. Mol. Biol. 219:277, 1991.)
Accordingly, the definition of class I and class II allele-specific HLA binding motifs, or class I or class II supermotifs allows identification of regions within a protein that are correlated with binding to particular HLA antigen(s).
Thus, by a process of HLA motif identification, candidates for epitope-based vaccines have been identified; such candidates can be further evaluated by HLA-peptide binding assays to determine binding affinity and/or the time period of association of the epitope and its corresponding HLA molecule. Additional confirmatory work can be performed to select, amongst these vaccine candidates, epitopes with preferred characteristics in terms of population coverage, and/or immunogenicity.
Various strategies can be utilized to evaluate cellular immunogenicity, including:
1) Evaluation of primary T cell cultures from normal individuals (see, e.g., Wentworth, P. A. et al., Mol. Immunol. 32:603, 1995; Celis, E. et al., Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al., J. Immunol. 158:1796, 1997; Kawashima, I. et al., Human Immunol. 59:1, 1998). This procedure involves the stimulation of peripheral blood lymphocytes (PBL) from normal subjects with a test peptide in the presence of antigen presenting cells in vitro over a period of several weeks. T cells specific for the peptide become activated during this time and are detected using, e.g., a lymphokine- or 51Cr-release assay involving peptide sensitized target cells.
2) Immunization of HLA transgenic mice (see, e.g., Wentworth, P. A. et al., J. Immunol. 26:97, 1996; Wentworth, P. A. et al., Int. Immunol. 8:651, 1996; Alexander, J. et al., J. Immunol. 159:4753, 1997). For example, in such methods peptides in incomplete Freund's adjuvant are administered subcutaneously to HLA transgenic mice. Several weeks following immunization, splenocytes are removed and cultured in vitro in the presence of test peptide for approximately one week. Peptide-specific T cells are detected using, e.g., a 51Cr-release assay involving peptide sensitized target cells and target cells expressing endogenously generated antigen.
3) Demonstration of recall T cell responses from immune individuals who have been either effectively vaccinated and/or from chronically ill patients (see, e.g., Rehermann, B. et al., J. Exp. Med. 181:1047, 1995; Doolan, D. L. et al., Immunity 7:97, 1997; Bertoni, R. et al., J. Clin. Invest. 100:503, 1997; Threlkeld, S. C. et al., J. Immunol. 159:1648, 1997; Diepolder, H. M. et al., J. Virol. 71:6011, 1997). Accordingly, recall responses are detected by culturing PBL from subjects that have been exposed to the antigen due to disease and thus have generated an immune response “naturally”, or from patients who were vaccinated against the antigen. PBL from subjects are cultured in vitro for 1-2 weeks in the presence of test peptide plus antigen presenting cells (APC) to allow activation of “memory” T cells, as compared to “naive” T cells. At the end of the culture period, T cell activity is detected using assays including 51Cr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.
VI.) 184P1E2 Transgenic Animals
Nucleic acids that encode a 184P1E2-related protein can also be used to generate either transgenic animals or “knock out” animals that, in turn, are useful in the development and screening of therapeutically useful reagents. In accordance with established techniques, cDNA encoding 184P1E2 can be used to clone genomic DNA that encodes 184P1E2. The cloned genomic sequences can then be used to generate transgenic animals containing cells that express DNA that encode 184P1E2. Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art and are described, for example, in U.S. Pat. No. 4,736,866 issued 12 Apr. 1988, and U.S. Pat. No. 4,870,009 issued 26 Sep. 1989. Typically, particular cells would be targeted for 184P1E2 transgene incorporation with tissue-specific enhancers.
Transgenic animals that include a copy of a transgene encoding 184P1E2 can be used to examine the effect of increased expression of DNA that encodes 184P1E2. Such animals can be used as tester animals for reagents thought to confer protection from, for example, pathological conditions associated with its overexpression. In accordance with this aspect of the invention, an animal is treated with a reagent and a reduced incidence of a pathological condition, compared to untreated animals that bear the transgene, would indicate a potential therapeutic intervention for the pathological condition.
Alternatively, non-human homologues of 184P1E2 can be used to construct a 184P1E2 “knock out” animal that has a defective or altered gene encoding 184P1E2 as a result of homologous recombination between the endogenous gene encoding 184P1E2 and altered genomic DNA encoding 184P1E2 introduced into an embryonic cell of the animal. For example, cDNA that encodes 184P1E2 can be used to clone genomic DNA encoding 184P1E2 in accordance with established techniques. A portion of the genomic DNA encoding 184P1E2 can be deleted or replaced with another gene, such as a gene encoding a selectable marker that can be used to monitor integration. Typically, several kilobases of unaltered flanking DNA (both at the 5′ and 3′ ends) are included in the vector (see, e.g., Thomas and Capecchi, Cell, 51:503 (1987) for a description of homologous recombination vectors). The vector is introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced DNA has homologously recombined with the endogenous DNA are selected (see, e.g., Li et al., Cell, 69:915 (1992)). The selected cells are then injected into a blastocyst of an animal (e.g., a mouse or rat) to form aggregation chimeras (see, e.g., Bradley, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152). A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal, and the embryo brought to term to create a “knock out” animal. Progeny harboring the homologously recombined DNA in their germ cells can be identified by standard techniques and used to breed animals in which all cells of the animal contain the homologously recombined DNA. Knock out animals can be characterized, for example, for their ability to defend against certain pathological conditions or for their development of pathological conditions due to absence of a 184P1E2 polypeptide.
VII.) Methods for the Detection of 184P1E2
Another aspect of the present invention relates to methods for detecting 184P1E2 polynucleotides and 184P1E2-related proteins, as well as methods for identifying a cell that expresses 184P1E2. The expression profile of 184P1E2 makes it a diagnostic marker for metastasized disease. Accordingly, the status of 184P1E2 gene products provides information useful for predicting a variety of factors including susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. As discussed in detail herein, the status of 184P1E2 gene products in patient samples can be analyzed by a variety protocols that are well known in the art including immunohistochemical analysis, the variety of Northern blotting techniques including in situ hybridization, RT-PCR analysis (for example on laser capture micro-dissected samples), Western blot analysis and tissue array analysis.
More particularly, the invention provides assays for the detection of 184P1E2 polynucleotides in a biological sample, such as serum, bone, prostate, and other tissues, urine, semen, cell preparations, and the like. Detectable 184P1E2 polynucleotides include, for example, a 184P1E2 gene or fragment thereof, 184P1E2 mRNA, alternative splice variant 184P1E2 mRNAs, and recombinant DNA or RNA molecules that contain a 184P1E2 polynucleotide. A number of methods for amplifying and/or detecting the presence of 184P1E2 polynucleotides are well known in the art and can be employed in the practice of this aspect of the invention.
In one embodiment, a method for detecting a 184P1E2 mRNA in a biological sample comprises producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using a 184P1E2 polynucleotides as sense and antisense primers to amplify 184P1E2 cDNAs therein; and detecting the presence of the amplified 184P1E2 cDNA. Optionally, the sequence of the amplified 184P1E2 cDNA can be determined.
In another embodiment, a method of detecting a 184P1E2 gene in a biological sample comprises first isolating genomic DNA from the sample; amplifying the isolated genomic DNA using 184P1E2 polynucleotides as sense and antisense primers; and detecting the presence of the amplified 184P1E2 gene. Any number of appropriate sense and antisense probe combinations can be designed from a 184P1E2 nucleotide sequence (see, e.g., FIG. 2) and used for this purpose.
The invention also provides assays for detecting the presence of a 184P1E2 protein in a tissue or other biological sample such as serum, semen, bone, prostate, urine, cell preparations, and the like. Methods for detecting a 184P1E2-related protein are also well known and include, for example, immunoprecipitation, immunohistochemical analysis, Western blot analysis, molecular binding assays, ELISA, ELIFA and the like. For example, a method of detecting the presence of a 184P1E2-related protein in a biological sample comprises first contacting the sample with a 184P1E2 antibody, a 184P1E2-reactive fragment thereof, or a recombinant protein containing an antigen binding region of a 184P1E2 antibody; and then detecting the binding of 184P1E2-related protein in the sample.
Methods for identifying a cell that expresses 184P1E2 are also within the scope of the invention. In one embodiment, an assay for identifying a cell that expresses a 184P1E2 gene comprises detecting the presence of 184P1E2 mRNA in the cell. Methods for the detection of particular mRNAs in cells are well known and include, for example, hybridization assays using complementary DNA probes (such as in situ hybridization using labeled 184P1E2 riboprobes, Northern blot and related techniques) and various nucleic acid amplification assays (such as RT-PCR using complementary primers specific for 184P1E2, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like). Alternatively, an assay for identifying a cell that expresses a 184P1E2 gene comprises detecting the presence of 184P1E2-related protein in the cell or secreted by the cell. Various methods for the detection of proteins are well known in the art and are employed for the detection of 184P1E2-related proteins and cells that express 184P1E2-related proteins.
184P1E2 expression analysis is also useful as a tool for identifying and evaluating agents that modulate 184P1E2 gene expression. For example, 184P1E2 expression is significantly upregulated in prostate cancer, and is expressed in cancers of the tissues listed in Table I. Identification of a molecule or biological agent that inhibits 184P1E2 expression or over-expression in cancer cells is of therapeutic value. For example, such an agent can be identified by using a screen that quantifies 184P1E2 expression by RT-PCR, nucleic acid hybridization or antibody binding.
VIII.) Methods for Monitoring the Status of 184P1E2-Related Genes and their Products
Oncogenesis is known to be a multistep process where cellular growth becomes progressively dysregulated and cells progress from a normal physiological state to precancerous and then cancerous states (see, e.g., Alers et al., Lab Invest. 77(5): 437-438 (1997) and Isaacs et al., Cancer Surv. 23: 19-32 (1995)). In this context, examining a biological sample for evidence of dysregulated cell growth (such as aberrant 184P1E2 expression in cancers) allows for early detection of such aberrant physiology, before a pathologic state such as cancer has progressed to a stage that therapeutic options are more limited and or the prognosis is worse. In such examinations, the status of 184P1E2 in a biological sample of interest can be compared, for example, to the status of 184P1E2 in a corresponding normal sample (e.g. a sample from that individual or alternatively another individual that is not affected by a pathology). An alteration in the status of 184P1E2 in the biological sample (as compared to the normal sample) provides evidence of dysregulated cellular growth. In addition to using a biological sample that is not affected by a pathology as a normal sample, one can also use a predetermined normative value such as a predetermined normal level of mRNA expression (see, e.g., Grever et al., J. Comp. Neurol. 1996 Dec. 9; 376(2): 306-14 and U.S. Pat. No. 5,837,501) to compare 184P1E2 status in a sample.
The term “status” in this context is used according to its art accepted meaning and refers to the condition or state of a gene and its products. Typically, skilled artisans use a number of parameters to evaluate the condition or state of a gene and its products. These include, but are not limited to the location of expressed gene products (including the location of 184P1E2 expressing cells) as well as the level, and biological activity of expressed gene products (such as 184P1E2 mRNA, polynucleotides and polypeptides). Typically, an alteration in the status of 184P1E2 comprises a change in the location of 184P1E2 and/or 184P1E2 expressing cells and/or an increase in 184P1E2 mRNA and/or protein expression.
184P1E2 status in a sample can be analyzed by a number of means well known in the art, including without limitation, immunohistochemical analysis, in situ hybridization, RT-PCR analysis on laser capture micro-dissected samples, Western blot analysis, and tissue array analysis. Typical protocols for evaluating the status of a 184P1E2 gene and gene products are found, for example in Ausubel et al. eds., 1995, Current Protocols In Molecular Biology, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting) and 18 (PCR Analysis). Thus, the status of 184P1E2 in a biological sample is evaluated by various methods utilized by skilled artisans including, but not limited to genomic Southern analysis (to examine, for example perturbations in a 184P1E2 gene), Northern analysis and/or PCR analysis of 184P1E2 mRNA (to examine, for example alterations in the polynucleotide sequences or expression levels of 184P1E2 mRNAs), and, Western and/or immunohistochemical analysis (to examine, for example alterations in polypeptide sequences, alterations in polypeptide localization within a sample, alterations in expression levels of 184P1E2 proteins and/or associations of 184P1E2 proteins with polypeptide binding partners). Detectable 184P1E2 polynucleotides include, for example, a 184P1E2 gene or fragment thereof, 184P1E2 mRNA, alternative splice variants, 184P1E2 mRNAs, and recombinant DNA or RNA molecules containing a 184P1E2 polynucleotide.
The expression profile of 184P1E2 makes it a diagnostic marker for local and/or metastasized disease, and provides information on the growth or oncogenic potential of a biological sample. In particular, the status of 184P1E2 provides information useful for predicting susceptibility to particular disease stages, progression, and/or tumor aggressiveness. The invention provides methods and assays for determining 184P1E2 status and diagnosing cancers that express 184P1E2, such as cancers of the tissues listed in Table 1. For example, because 184P1E2 mRNA is so highly expressed in prostate and other cancers relative to normal prostate tissue, assays that evaluate the levels of 184P1E2 mRNA transcripts or proteins in a biological sample can be used to diagnose a disease associated with 184P1E2 dysregulation, and can provide prognostic information useful in defining appropriate therapeutic options.
The expression status of 184P1E2 provides information including the presence, stage and location of dysplastic, precancerous and cancerous cells, predicting susceptibility to various stages of disease, and/or for gauging tumor aggressiveness. Moreover, the expression profile makes it useful as an imaging reagent for metastasized disease. Consequently, an aspect of the invention is directed to the various molecular prognostic and diagnostic methods for examining the status of 184P1E2 in biological samples such as those from individuals suffering from, or suspected of suffering from a pathology characterized by dysregulated cellular growth, such as cancer.
As described above, the status of 184P1E2 in a biological sample can be examined by a number of well-known procedures in the art. For example, the status of 184P1E2 in a biological sample taken from a specific location in the body can be examined by evaluating the sample for the presence or absence of 184P1E2 expressing cells (e.g. those that express 184P1E2 mRNAs or proteins). This examination can provide evidence of dysregulated cellular growth, for example, when 184P1E2-expressing cells are found in a biological sample that does not normally contain such cells (such as a lymph node), because such alterations in the status of 184P1E2 in a biological sample are often associated with dysregulated cellular growth. Specifically, one indicator of dysregulated cellular growth is the metastases of cancer cells from an organ of origin (such as the prostate) to a different area of the body (such as a lymph node). In this context, evidence of dysregulated cellular growth is important for example because occult lymph node metastases can be detected in a substantial proportion of patients with prostate cancer, and such metastases are associated with known predictors of disease progression (see, e.g., Murphy et al., Prostate 42(4): 315-317 (2000);Su et al., Semin. Surg. Oncol. 18(1): 17-28 (2000) and Freeman et al., J Urol 1995 August 154(2 Pt 1):474-8).
In one aspect, the invention provides methods for monitoring 184P1E2 gene products by determining the status of 184P1E2 gene products expressed by cells from an individual suspected of having a disease associated with dysregulated cell growth (such as hyperplasia or cancer) and then comparing the status so determined to the status of 184P1E2 gene products in a corresponding normal sample. The presence of aberrant 184P1E2 gene products in the test sample relative to the normal sample provides an indication of the presence of dysregulated cell growth within the cells of the individual.
In another aspect, the invention provides assays useful in determining the presence of cancer in an individual, comprising detecting a significant increase in 184P1E2 mRNA or protein expression in a test cell or tissue sample relative to expression levels in the corresponding normal cell or tissue. The presence of 184P1E2 mRNA can, for example, be evaluated in tissues including but not limited to those listed in Table I. The presence of significant 184P1E2 expression in any of these tissues is useful to indicate the emergence, presence and/or severity of a cancer, since the corresponding normal tissues do not express 184P1E2 mRNA or express it at lower levels.
In a related embodiment, 184P1E2 status is determined at the protein level rather than at the nucleic acid level. For example, such a method comprises determining the level of 184P1E2 protein expressed by cells in a test tissue sample and comparing the level so determined to the level of 184P1E2 expressed in a corresponding normal sample. In one embodiment, the presence of 184P1E2 protein is evaluated, for example, using immunohistochemical methods. 184P1E2 antibodies or binding partners capable of detecting 184P1E2 protein expression are used in a variety of assay formats well known in the art for this purpose.
In a further embodiment, one can evaluate the status of 184P1E2 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules. These perturbations can include insertions, deletions, substitutions and the like. Such evaluations are useful because perturbations in the nucleotide and amino acid sequences are observed in a large number of proteins associated with a growth dysregulated phenotype (see, e.g., Marrogi et al., 1999, J. Cutan. Pathol. 26(8):369-378). For example, a mutation in the sequence of 184P1E2 may be indicative of the presence or promotion of a tumor. Such assays therefore have diagnostic and predictive value where a mutation in 184P1E2 indicates a potential loss of function or increase in tumor growth.
A wide variety of assays for observing perturbations in nucleotide and amino acid sequences are well known in the art. For example, the size and structure of nucleic acid or amino acid sequences of 184P1E2 gene products are observed by the Northern, Southern, Western, PCR and DNA sequencing protocols discussed herein. In addition, other methods for observing perturbations in nucleotide and amino acid sequences such as single strand conformation polymorphism analysis are well known in the art (see, e.g., U.S. Pat. No. 5,382,510 issued 7 Sep. 1999, and U.S. Pat. No. 5,952,170 issued 17 Jan. 1995).
Additionally, one can examine the methylation status of a 184P1E2 gene in a biological sample. Aberrant demethylation and/or hypermethylation of CpG islands in gene 5′ regulatory regions frequently occurs in immortalized and transformed cells, and can result in altered expression of various genes. For example, promoter hypermethylation of the pi-class glutathione S-transferase (a protein expressed in normal prostate but not expressed in >90% of prostate carcinomas) appears to permanently silence transcription of this gene and is the most frequently detected genomic alteration in prostate carcinomas (De Marzo et al., Am. J. Pathol. 155(6): 1985-1992 (1999)). In addition, this alteration is present in at least 70% of cases of high-grade prostatic intraepithelial neoplasia (PIN) (Brooks et al., Cancer Epidemiol. Biomarkers Prev., 1998, 7:531-536). In another example, expression of the LAGE-I tumor specific gene (which is not expressed in normal prostate but is expressed in 25-50% of prostate cancers) is induced by deoxy-azacytidine in lymphoblastoid cells, suggesting that tumoral expression is due to demethylation (Lethe et al., Int. J. Cancer 76(6): 903-908 (1998)). A variety of assays for examining methylation status of a gene are well known in the art. For example, one can utilize, in Southern hybridization approaches, methylation-sensitive restriction enzymes that cannot cleave sequences that contain methylated CpG sites to assess the methylation status of CpG islands. In addition, MSP (methylation specific PCR) can rapidly profile the methylation status of all the CpG sites present in a CpG island of a given gene. This procedure involves initial modification of DNA by sodium bisulfite (which will convert all unmethylated cytosines to uracil) followed by amplification using primers specific for methylated versus unmethylated DNA. Protocols involving methylation interference can also be found for example in Current Protocols In Molecular Biology, Unit 12, Frederick M. Ausubel et al. eds., 1995.
Gene amplification is an additional method for assessing the status of 184P1E2. Gene amplification is measured in a sample directly, for example, by conventional Southern blotting or Northern blotting to quantitate the transcription of mRNA (Thomas, 1980, Proc. Natl. Acad. Sci. USA, 77:5201-5205), dot blotting (DNA analysis), or in situ hybridization, using an appropriately labeled probe, based on the sequences provided herein. Alternatively, antibodies are employed that recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. The antibodies in turn are labeled and the assay carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected.
Biopsied tissue or peripheral blood can be conveniently assayed for the presence of cancer cells using for example, Northern, dot blot or RT-PCR analysis to detect 184P1E2 expression. The presence of RT-PCR amplifiable 184P1E2 mRNA provides an indication of the presence of cancer. RT-PCR assays are well known in the art. RT-PCR detection assays for tumor cells in peripheral blood are currently being evaluated for use in the diagnosis and management of a number of human solid tumors. In the prostate cancer field, these include RT-PCR assays for the detection of cells expressing PSA and PSM (Verkaik et al., 1997, Urol. Res. 25:373-384; Ghossein et al., 1995, J. Clin. Oncol. 13:1195-2000; Heston et al., 1995, Clin. Chem. 41:1687-1688).
A further aspect of the invention is an assessment of the susceptibility that an individual has for developing cancer. In one embodiment, a method for predicting susceptibility to cancer comprises detecting 184P1E2 mRNA or 184P1E2 protein in a tissue sample, its presence indicating susceptibility to cancer, wherein the degree of 184P1E2 mRNA expression correlates to the degree of susceptibility. In a specific embodiment, the presence of 184P1E2 in prostate or other tissue is examined, with the presence of 184P1E2 in the sample providing an indication of prostate cancer susceptibility (or the emergence or existence of a prostate tumor). Similarly, one can evaluate the integrity 184P1E2 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations in 184P1E2 gene products in the sample is an indication of cancer susceptibility (or the emergence or existence of a tumor).
The invention also comprises methods for gauging tumor aggressiveness. In one embodiment, a method for gauging aggressiveness of a tumor comprises determining the level of 184P1E2 mRNA or 184P1E2 protein expressed by tumor cells, comparing the level so determined to the level of 184P1E2 mRNA or 184P1E2 protein expressed in a corresponding normal tissue taken from the same individual or a normal tissue reference sample, wherein the degree of 184P1E2 mRNA or 184P1E2 protein expression in the tumor sample relative to the normal sample indicates the degree of aggressiveness. In a specific embodiment, aggressiveness of a tumor is evaluated by determining the extent to which 184P1E2 is expressed in the tumor cells, with higher expression levels indicating more aggressive tumors. Another embodiment is the evaluation of the integrity of 184P1E2 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations indicates more aggressive tumors.
Another embodiment of the invention is directed to methods for observing the progression of a malignancy in an individual over time. In one embodiment, methods for observing the progression of a malignancy in an individual over time comprise determining the level of 184P1E2 mRNA or 184P1E2 protein expressed by cells in a sample of the tumor, comparing the level so determined to the level of 184P1E2 mRNA or 184P1E2 protein expressed in an equivalent tissue sample taken from the same individual at a different time, wherein the degree of 184P1E2 mRNA or 184P1E2 protein expression in the tumor sample over time provides information on the progression of the cancer. In a specific embodiment, the progression of a cancer is evaluated by determining 184P1E2 expression in the tumor cells over time, where increased expression over time indicates a progression of the cancer. Also, one can evaluate the integrity 184P1E2 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like, where the presence of one or more perturbations indicates a progression of the cancer.
The above diagnostic approaches can be combined with any one of a wide variety of prognostic and diagnostic protocols known in the art. For example, another embodiment of the invention is directed to methods for observing a coincidence between the expression of 184P1E2 gene and 184P1E2 gene products (or perturbations in 184P1E2 gene and 184P1E2 gene products) and a factor that is associated with malignancy, as a means for diagnosing and prognosticating the status of a tissue sample. A wide variety of factors associated with malignancy can be utilized, such as the expression of genes associated with malignancy (e.g. PSA, PSCA and PSM expression for prostate cancer etc.) as well as gross cytological observations (see, e.g., Bocking et al., 1984, Anal. Quant. Cytol. 6(2):74-88; Epstein, 1995, Hum. Pathol. 26(2):223-9; Thorson et al., 1998, Mod. Pathol. 11(6):543-51; Baisden et al., 1999, Am. J. Surg. Pathol. 23(8):918-24). Methods for observing a coincidence between the expression of 184P1E2 gene and 184P1E2 gene products (or perturbations in 184P1E2 gene and 184P1E2 gene products) and another factor that is associated with malignancy are useful, for example, because the presence of a set of specific factors that coincide with disease provides information crucial for diagnosing and prognosticating the status of a tissue sample.
In one embodiment, methods for observing a coincidence between the expression of 184P1E2 gene and 184P1E2 gene products (or perturbations in 184P1E2 gene and 184P1E2 gene products) and another factor associated with malignancy entails detecting the overexpression of 184P1E2 mRNA or protein in a tissue sample, detecting the overexpression of PSA mRNA or protein in a tissue sample (or PSCA or PSM expression), and observing a coincidence of 184P1E2 mRNA or protein and PSA mRNA or protein overexpression (or PSCA or PSM expression). In a specific embodiment, the expression of 184P1E2 and PSA mRNA in prostate tissue is examined, where the coincidence of 184P1E2 and PSA mRNA overexpression in the sample indicates the existence of prostate cancer, prostate cancer susceptibility or the emergence or status of a prostate tumor.
Methods for detecting and quantifying the expression of 184P1E2 mRNA or protein are described herein, and standard nucleic acid and protein detection and quantification technologies are well known in the art. Standard methods for the detection and quantification of 184P1E2 mRNA include in situ hybridization using labeled 184P1E2 riboprobes, Northern blot and related techniques using 184P1E2 polynucleotide probes, RT-PCR analysis using primers specific for 184P1E2, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like. In a specific embodiment, semi-quantitative RT-PCR is used to detect and quantify 184P1E2 mRNA expression. Any number of primers capable of amplifying 184P1E2 can be used for this purpose, including but not limited to the various primer sets specifically described herein. In a specific embodiment, polyclonal or monoclonal antibodies specifically reactive with the wild-type 184P1E2 protein can be used in an immunohistochemical assay of biopsied tissue.
IX.) Identification of Molecules that Interact with 184P1E2
The 184P1E2 protein and nucleic acid sequences disclosed herein allow a skilled artisan to identify proteins, small molecules and other agents that interact with 184P1E2, as well as pathways activated by 184P1E2 via any one of a variety of art accepted protocols. For example, one can utilize one of the so-called interaction trap systems (also referred to as the “two-hybrid assay”). In such systems, molecules interact and reconstitute a transcription factor which directs expression of a reporter gene, whereupon the expression of the reporter gene is assayed. Other systems identify protein-protein interactions in vivo through reconstitution of a eukaryotic transcriptional activator, see, e.g., U.S. Pat. No. 5,955,280 issued 21 Sep. 1999, U.S. Pat. No. 5,925,523 issued 20 Jul. 1999, U.S. Pat. No. 5,846,722 issued 8 Dec. 1998 and U.S. Pat. No. 6,004,746 issued 21 Dec. 1999. Algorithms are also available in the art for genome-based predictions of protein function (see, e.g., Marcotte, et al., Nature 402: 4 Nov. 1999, 83-86).
Alternatively one can screen peptide libraries to identify molecules that interact with 184P1E2 protein sequences. In such methods, peptides that bind to 184P1E2 are identified by screening libraries that encode a random or controlled collection of amino acids. Peptides encoded by the libraries are expressed as fusion proteins of bacteriophage coat proteins, the bacteriophage particles are then screened against the 184P1E2 protein(s).
Accordingly, peptides having a wide variety of uses, such as therapeutic, prognostic or diagnostic reagents, are thus identified without any prior information on the structure of the expected ligand or receptor molecule. Typical peptide libraries and screening methods that can be used to identify molecules that interact with 184P1E2 protein sequences are disclosed for example in U.S. Pat. No. 5,723,286 issued 3 Mar. 1998 and U.S. Pat. No. 5,733,731 issued 31 Mar. 1998.
Alternatively, cell lines that express 184P1E2 are used to identify protein-protein interactions mediated by 184P1E2. Such interactions can be examined using immunoprecipitation techniques (see, e.g., Hamilton B. J., et al. Biochem. Biophys. Res. Commun. 1999, 261:646-51). 184P1E2 protein can be immunoprecipitated from 184P1E2-expressing cell lines using anti-184P1E2 antibodies. Alternatively, antibodies against His-tag can be used in a cell line engineered to express fusions of 184P1E2 and a His-tag (vectors mentioned above). The immunoprecipitated complex can be examined for protein association by procedures such as Western blotting, 35S-methionine labeling of proteins, protein microsequencing, silver staining and two-dimensional gel electrophoresis.
Small molecules and ligands that interact with 184P1E2 can be identified through related embodiments of such screening assays. For example, small molecules can be identified that interfere with protein function, including molecules that interfere with 184P1E2's ability to mediate phosphorylation and de-phosphorylation, interaction with DNA or RNA molecules as an indication of regulation of cell cycles, second messenger signaling or tumorigenesis. Similarly, small molecules that modulate 184P1E2-related ion channel, protein pump, or cell communication functions are identified and used to treat patients that have a cancer that expresses 184P1E2 (see, e.g., Hille, B., Ionic Channels of Excitable Membranes 2nd Ed., Sinauer Assoc., Sunderland, Mass., 1992). Moreover, ligands that regulate 184P1E2 function can be identified based on their ability to bind 184P1E2 and activate a reporter construct. Typical methods are discussed for example in U.S. Pat. No. 5,928,868 issued 27 Jul. 1999, and include methods for forming hybrid ligands in which at least one ligand is a small molecule. In an illustrative embodiment, cells engineered to express a fusion protein of 184P1E2 and a DNA-binding protein are used to co-express a fusion protein of a hybrid ligand/small molecule and a cDNA library transcriptional activator protein. The cells further contain a reporter gene, the expression of which is conditioned on the proximity of the first and second fusion proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cells that express the reporter gene are selected and the unknown small molecule or the unknown ligand is identified. This method provides a means of identifying modulators which activate or inhibit 184P1E2.
An embodiment of this invention comprises a method of screening for a molecule that interacts with a 184P1E2 amino acid sequence shown in FIG. 2 or FIG. 3, comprising the steps of contacting a population of molecules with a 184P1E2 amino acid sequence, allowing the population of molecules and the 184P1E2 amino acid sequence to interact under conditions that facilitate an interaction, determining the presence of a molecule that interacts with the 184P1E2 amino acid sequence, and then separating molecules that do not interact with the 184P1E2 amino acid sequence from molecules that do. In a specific embodiment, the method further comprises purifying, characterizing and identifying a molecule that interacts with the 184P1E2 amino acid sequence. The identified molecule can be used to modulate a function performed by 184P1E2. In a preferred embodiment, the 184P1E2 amino acid sequence is contacted with a library of peptides.
X.) Therapeutic Methods and Compositions
The identification of 184P1E2 as a protein that is normally expressed in a restricted set of tissues, but which is also expressed in prostate and other cancers, opens a number of therapeutic approaches to the treatment of such cancers. As contemplated herein, 184P1E2 functions as a transcription factor involved in activating tumor-promoting genes or repressing genes that block tumorigenesis.
Accordingly, therapeutic approaches that inhibit the activity of a 184P1E2 protein are useful for patients suffering from a cancer that expresses 184P1E2. These therapeutic approaches generally fall into two classes. One class comprises various methods for inhibiting the binding or association of a 184P1E2 protein with its binding partner or with other proteins. Another class comprises a variety of methods for inhibiting the transcription of a 184P1E2 gene or translation of 184P1E2 mRNA.
X.A.) Anti-Cancer Vaccines
The invention provides cancer vaccines comprising a 184P1E2-related protein or 184P1E2-related nucleic acid. In view of the expression of 184P1E2, cancer vaccines prevent and/or treat 184P1E2-expressing cancers with minimal or no effects on non-target tissues. The use of a tumor antigen in a vaccine that generates humoral and/or cell-mediated immune responses as anti-cancer therapy is well known in the art and has been employed in prostate cancer using human PSMA and rodent PAP immunogens (Hodge et al., 1995, Int. J. Cancer 63:231-237; Fong et al., 1997, J. Immunol. 159:3113-3117).
Such methods can be readily practiced by employing a 184P1E2-related protein, or a 184P1E2-encoding nucleic acid molecule and recombinant vectors capable of expressing and presenting the 184P1E2 immunogen (which typically comprises a number of antibody or T cell epitopes). Skilled artisans understand that a wide variety of vaccine systems for delivery of immunoreactive epitopes are known in the art (see, e.g., Heryln et al., Ann Med 1999 February 31(1):66-78; Maruyama et al., Cancer Immunol Immunother 2000 June 49(3):123-32) Briefly, such methods of generating an immune response (e.g. humoral and/or cell-mediated) in a mammal, comprise the steps of: exposing the mammal's immune system to an immunoreactive epitope (e.g. an epitope present in a 184P1E2 protein shown in FIG. 3 or analog or homolog thereof) so that the mammal generates an immune response that is specific for that epitope (e.g. generates antibodies that specifically recognize that epitope). In a preferred method, a 184P1E2 immunogen contains a biological motif, see e.g., Tables V-XVIII and XXII-LI, or a peptide of a size range from 184P1E2 indicated in FIG. 5, FIG. 6, FIG. 7, FIG. 8, and FIG. 9.
The entire 184P1E2 protein, immunogenic regions or epitopes thereof can be combined and delivered by various means. Such vaccine compositions can include, for example, lipopeptides (e.g., Vitiello, A. et al., J. Clin. Invest. 95:341, 1995), peptide compositions encapsulated in poly(DL-lactide-co-glycolide) (“PLG”) microspheres (see, e.g., Eldridge, et al., Molec. Immunol. 28:287-294, 1991: Alonso et al., Vaccine 12:299-306, 1994; Jones et al., Vaccine 13:675-681, 1995), peptide compositions contained in immune stimulating complexes (ISCOMS) (see, e.g., Takahashi et al., Nature 344:873-875, 1990; Hu et al., Clin Exp Immunol. 113:235-243, 1998), multiple antigen peptide systems (MAPs) (see e.g., Tam, J. P., Proc. Natl. Acad. Sci. U.S.A. 85:5409-5413, 1988; Tam, J. P., J. Immunol. Methods 196:17-32, 1996), peptides formulated as multivalent peptides; peptides for use in ballistic delivery systems, typically crystallized peptides, viral delivery vectors (Perkus, M. E. et al., In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 379, 1996; Chakrabarti, S. et al., Nature 320:535, 1986; Hu, S. L. et al., Nature 320:537, 1986; Kieny, M.-P. et al., AIDS Bio/Technology 4:790, 1986; Top, F. H. et al., J. Infect. Dis. 124:148, 1971; Chanda, P. K. et al., Virology 175:535, 1990), particles of viral or synthetic origin (e.g., Kofler, N. et al., J. Immunol. Methods. 192:25, 1996; Eldridge, J. H. et al., Sem. Hematol. 30:16, 1993; Falo, L. D., Jr. et al., Nature Med. 7:649, 1995), adjuvants (Warren, H. S., Vogel, F. R., and Chedid, L. A. Annu. Rev. Immunol. 4:369, 1986; Gupta, R. K. et al., Vaccine 11:293, 1993), liposomes (Reddy, R. et al., J. Immunol. 148:1585, 1992; Rock, K. L., Immunol. Today 17:131, 1996), or, naked or particle absorbed cDNA (Ulmer, J. B. et al., Science 259:1745, 1993; Robinson, H. L., Hunt, L. A., and Webster, R. G., Vaccine 11:957, 1993; Shiver, J. W. et al., In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 423, 1996; Cease, K. B., and Berzofsky, J. A., Annu. Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et al., Sem. Hematol. 30:16, 1993). Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Mass.) may also be used.
In patients with 184P1E2-associated cancer, the vaccine compositions of the invention can also be used in conjunction with other treatments used for cancer, e.g., surgery, chemotherapy, drug therapies, radiation therapies, etc. including use in combination with immune adjuvants such as IL-2, IL-12, GM-CSF, and the like.
Cellular Vaccines:
CTL epitopes can be determined using specific algorithms to identify peptides within 184P1E2 protein that bind corresponding HLA alleles (see e.g., Table IV; EpimerTM and EpimatrixTM, Brown University, BIMAS and INTERNET site SYFPEITHI. In a preferred embodiment, a 184P1E2 immunogen contains one or more amino acid sequences identified using techniques well known in the art, such as the sequences shown in Tables V-XVIII and XXII-LI or a peptide of 8, 9, 10 or 11 amino acids specified by an HLA Class I motif/supermotif (e.g., Table IV (A), Table IV (D), or Table IV (E)) and/or a peptide of at least 9 amino acids that comprises an HLA Class II motif/supermotif (e.g., Table IV (B) or Table IV (C)). As is appreciated in the art, the HLA Class I binding groove is essentially closed ended so that peptides of only a particular size range can fit into the groove and be bound, generally HLA Class I epitopes are 8, 9, 10, or 11 amino acids long. In contrast, the HLA Class II binding groove is essentially open ended; therefore a peptide of about 9 or more amino acids can be bound by an HLA Class I molecule. Due to the binding groove differences between HLA Class I and II, HLA Class I motifs are length specific, i.e., position two of a Class I motif is the second amino acid in an amino to carboxyl direction of the peptide. The amino acid positions in a Class II motif are relative only to each other, not the overall peptide, i.e., additional amino acids can be attached to the amino and/or carboxyl termini of a motif-bearing sequence. HLA Class II epitopes are often 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long, or longer than 25 amino acids.
Antibody-Based Vaccines
A wide variety of methods for generating an immune response in a mammal are known in the art (for example as the first step in the generation of hybridomas). Methods of generating an immune response in a mammal comprise exposing the mammal's immune system to an immunogenic epitope on a protein (e.g. a 184P1E2 protein) so that an immune response is generated. A typical embodiment consists of a method for generating an immune response to 184P1E2 in a host, by contacting the host with a sufficient amount of at least one 184P1E2 B cell or cytotoxic T-cell epitope or analog thereof; and at least one periodic interval thereafter re-contacting the host with the 184P1E2 B cell or cytotoxic T-cell epitope or analog thereof. A specific embodiment consists of a method of generating an immune response against a 184P1E2-related protein or a man-made multiepitopic peptide comprising: administering 184P1E2 immunogen (e.g. a 184P1E2 protein or a peptide fragment thereof, a 184P1E2 fusion protein or analog etc.) in a vaccine preparation to a human or another mammal. Typically, such vaccine preparations further contain a suitable adjuvant (see, e.g., U.S. Pat. No. 6,146,635) or a universal helper epitope such as a PADRETM peptide (Epimmune Inc., San Diego, Calif.; see, e.g., Alexander et al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., Immunity 1994 1(9): 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92). An alternative method comprises generating an immune response in an individual against a 184P1E2 immunogen by: administering in vivo to muscle or skin of the individual's body a DNA molecule that comprises a DNA sequence that encodes a 184P1E2 immunogen, the DNA sequence operatively linked to regulatory sequences which control the expression of the DNA sequence; wherein the DNA molecule is taken up by cells, the DNA sequence is expressed in the cells and an immune response is generated against the immunogen (see, e.g., U.S. Pat. No. 5,962,428). Optionally a genetic vaccine facilitator such as anionic lipids; saponins; lectins; estrogenic compounds; hydroxylated lower alkyls; dimethyl sulfoxide; and urea is also administered. In addition, an antiidiotypic antibody can be administered that mimics 184P1E2, in order to generate a response to the target antigen.
Nucleic Acid Vaccines:
Vaccine compositions of the invention include nucleic acid-mediated modalities. DNA or RNA that encode protein(s) of the invention can be administered to a patient. Genetic immunization methods can be employed to generate prophylactic or therapeutic humoral and cellular immune responses directed against cancer cells expressing 184P1E2. Constructs comprising DNA encoding a 184P1E2-related protein/immunogen and appropriate regulatory sequences can be injected directly into muscle or skin of an individual, such that the cells of the muscle or skin take-up the construct and express the encoded 184P1E2 protein/immunogen. Alternatively, a vaccine comprises a 184P1E2-related protein. Expression of the 184P1E2-related protein immunogen results in the generation of prophylactic or therapeutic humoral and cellular immunity against cells that bear a 184P1E2 protein. Various prophylactic and therapeutic genetic immunization techniques known in the art can be used (for review, see information and references published on the INTERNET). Nucleic acid-based delivery is described, for instance, in Wolff, et. al., Science 247:1465 (1990) as well as U.S. Pat. Nos. 5,580,859; 5,589,466; 5,804,566; 5,739,118; 5,736,524; 5,679,647; WO 98/04720. Examples of DNA-based delivery technologies include “naked DNA”, facilitated (bupivicaine, polymers, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated (“gene gun”) or pressure-mediated delivery (see, e.g., U.S. Pat. No. 5,922,687).
For therapeutic or prophylactic immunization purposes, proteins of the invention can be expressed via viral or bacterial vectors. Various viral gene delivery systems that can be used in the practice of the invention include, but are not limited to, vaccinia, fowlpox, canarypox, adenovirus, influenza, poliovirus, adeno-associated virus, lentivirus, and sindbis virus (see, e.g., Restifo, 1996, Curr. Opin. Immunol. 8:658-663; Tsang et al. J. Natl Cancer Inst. 87:982-990 (1995)). Non-viral delivery systems can also be employed by introducing naked DNA encoding a 184P1E2-related protein into the patient (e.g., intramuscularly or intradermally) to induce an anti-tumor response.
Vaccinia virus is used, for example, as a vector to express nucleotide sequences that encode the peptides of the invention. Upon introduction into a host, the recombinant vaccinia virus expresses the protein immunogenic peptide, and thereby elicits a host immune response. Vaccinia vectors and methods useful in immunization protocols are described in, e.g., U.S. Pat. No. 4,722,848. Another vector is BCG (Bacille Calmette Guerin). BCG vectors are described in Stover et al., Nature 351:456-460 (1991). A wide variety of other vectors useful for therapeutic administration or immunization of the peptides of the invention, e.g. adeno and adeno-associated virus vectors, retroviral vectors, Salmonella typhi vectors, detoxified anthrax toxin vectors, and the like, will be apparent to those skilled in the art from the description herein.
Thus, gene delivery systems are used to deliver a 184P1E2-related nucleic acid molecule. In one embodiment, the full-length human 184P1E2 cDNA is employed. In another embodiment, 184P1E2 nucleic acid molecules encoding specific cytotoxic T lymphocyte (CTL) and/or antibody epitopes are employed.
Ex Vivo Vaccines
Various ex vivo strategies can also be employed to generate an immune response. One approach involves the use of antigen presenting cells (APCs) such as dendritic cells (DC) to present 184P1E2 antigen to a patient's immune system. Dendritic cells express MHC class I and II molecules, B7 co-stimulator, and IL-12, and are thus highly specialized antigen presenting cells. In prostate cancer, autologous dendritic cells pulsed with peptides of the prostate-specific membrane antigen (PSMA) are being used in a Phase I clinical trial to stimulate prostate cancer patients' immune systems (Tjoa et al., 1996, Prostate 28:65-69; Murphy et al., 1996, Prostate 29:371-380). Thus, dendritic cells can be used to present 184P1E2 peptides to T cells in the context of MHC class I or II molecules. In one embodiment, autologous dendritic cells are pulsed with 184P1E2 peptides capable of binding to MHC class I and/or class II molecules. In another embodiment, dendritic cells are pulsed with the complete 184P1E2 protein. Yet another embodiment involves engineering the overexpression of a 184P1E2 gene in dendritic cells using various implementing vectors known in the art, such as adenovirus (Arthur et al., 1997, Cancer Gene Ther. 4:17-25), retrovirus (Henderson et al., 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-associated virus, DNA transfection (Ribas et al., 1997, Cancer Res. 57:2865-2869), or tumor-derived RNA transfection (Ashley et al., 1997, J. Exp. Med. 186:1177-1182). Cells that express 184P1E2 can also be engineered to express immune modulators, such as GM-CSF, and used as immunizing agents.
X.B.) 184P1E2 as a Target for Antibody-Based Therapy
184P1E2 is an attractive target for antibody-based therapeutic strategies. A number of antibody strategies are known in the art for targeting both extracellular and intracellular molecules (see, e.g., complement and ADCC mediated killing as well as the use of intrabodies). Because 184P1E2 is expressed by cancer cells of various lineages relative to corresponding normal cells, systemic administration of 184P1E2-immunoreactive compositions are prepared that exhibit excellent sensitivity without toxic, non-specific and/or non-target effects caused by binding of the immunoreactive composition to non-target organs and tissues. Antibodies specifically reactive with domains of 184P1E2 are useful to treat 184P1E2-expressing cancers systemically, either as conjugates with a toxin or therapeutic agent, or as naked antibodies capable of inhibiting cell proliferation or function.
184P1E2 antibodies can be introduced into a patient such that the antibody binds to 184P1E2 and modulates a function, such as an interaction with a binding partner, and consequently mediates destruction of the tumor cells and/or inhibits the growth of the tumor cells. Mechanisms by which such antibodies exert a therapeutic effect can include complement-mediated cytolysis, antibody-dependent cellular cytotoxicity, modulation of the physiological function of 184P1E2, inhibition of ligand binding or signal transduction pathways, modulation of tumor cell differentiation, alteration of tumor angiogenesis factor profiles, and/or apoptosis.
Those skilled in the art understand that antibodies can be used to specifically target and bind immunogenic molecules such as an immunogenic region of a 184P1E2 sequence shown in FIG. 2 or FIG. 3. In addition, skilled artisans understand that it is routine to conjugate antibodies to cytotoxic agents (see, e.g., Slevers et al. Blood 93:11 3678-3684 (Jun. 1, 1999)). When cytotoxic and/or therapeutic agents are delivered directly to cells, such as by conjugating them to antibodies specific for a molecule expressed by that cell (e.g. 184P1E2), the cytotoxic agent will exert its known biological effect (i.e. cytotoxicity) on those cells.
A wide variety of compositions and methods for using antibody-cytotoxic agent conjugates to kill cells are known in the art. In the context of cancers, typical methods entail administering to an animal having a tumor a biologically effective amount of a conjugate comprising a selected cytotoxic and/or therapeutic agent linked to a targeting agent (e.g. an anti-184P1E2 antibody) that binds to a marker (e.g. 184P1E2) expressed, accessible to binding or localized on the cell surfaces. A typical embodiment is a method of delivering a cytotoxic and/or therapeutic agent to a cell expressing 184P1E2, comprising conjugating the cytotoxic agent to an antibody that immunospecifically binds to a 184P1E2 epitope, and, exposing the cell to the antibody-agent conjugate. Another illustrative embodiment is a method of treating an individual suspected of suffering from metastasized cancer, comprising a step of administering parenterally to said individual a pharmaceutical composition comprising a therapeutically effective amount of an antibody conjugated to a cytotoxic and/or therapeutic agent.
Cancer immunotherapy using anti-184P1E2 antibodies can be done in accordance with various approaches that have been successfully employed in the treatment of other types of cancer, including but not limited to colon cancer (Arlen et al., 1998, Crit. Rev. Immunol. 18:133-138), multiple myeloma (Ozaki et al., 1997, Blood 90:3179-3186, Tsunenari et al., 1997, Blood 90:2437-2444), gastric cancer (Kasprzyk et al., 1992, Cancer Res. 52:2771-2776), B-cell lymphoma (Funakoshi et al., 1996, J. Immunother. Emphasis Tumor Immunol. 19:93-101), leukemia (Zhong et al., 1996, Leuk. Res. 20:581-589), colorectal cancer (Moun et al., 1994, Cancer Res. 54:6160-6166; Velders et al., 1995, Cancer Res. 55:4398-4403), and breast cancer (Shepard et al., 1991, J. Clin. Immunol. 11:117-127). Some therapeutic approaches involve conjugation of naked antibody to a toxin or radioisotope, such as the conjugation of Y91 or I131 to anti-CD20 antibodies (e.g., ZevalinTM, IDEC Pharmaceuticals Corp. or BexxarTM, Coulter Pharmaceuticals), while others involve co-administration of antibodies and other therapeutic agents, such as HerceptinTM (trastuzumab) with paclitaxel (Genentech, Inc.). The antibodies can be conjugated to a therapeutic agent. To treat prostate cancer, for example, 184P1E2 antibodies can be administered in conjunction with radiation, chemotherapy or hormone ablation. Also, antibodies can be conjugated to a toxin such as calicheamicin (e.g., MylotargTM, Wyeth-Ayerst, Madison, N.J., a recombinant humanized IgG4 kappa antibody conjugated to antitumor antibiotic calicheamicin) or a maytansinoid (e.g., taxane-based Tumor-Activated Prodrug, TAP, platform, ImmunoGen, Cambridge, Mass., also see e.g., U.S. Pat. No. 5,416,064).
Although 184P1E2 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemotherapeutic agent very well. Fan et al. (Cancer Res. 53:4637-4642, 1993), Prewett et al. (International J. of Onco. 9:217-224, 1996), and Hancock et al. (Cancer Res. 51:4575-4580, 1991) describe the use of various antibodies together with chemotherapeutic agents.
Although 184P1E2 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemotherapeutic agent very well.
Cancer patients can be evaluated for the presence and level of 184P1E2 expression, preferably using immunohistochemical assessments of tumor tissue, quantitative 184P1E2 imaging, or other techniques that reliably indicate the presence and degree of 184P1E2 expression. Immunohistochemical analysis of tumor biopsies or surgical specimens is preferred for this purpose. Methods for immunohistochemical analysis of tumor tissues are well known in the art.
Anti-184P1E2 monoclonal antibodies that treat prostate and other cancers include those that initiate a potent immune response against the tumor or those that are directly cytotoxic. In this regard, anti-184P1E2 monoclonal antibodies (mAbs) can elicit tumor cell lysis by either complement-mediated or antibody-dependent cell cytotoxicity (ADCC) mechanisms, both of which require an intact Fc portion of the immunoglobulin molecule for interaction with effector cell Fc receptor sites on complement proteins. In addition, anti-184P1E2 mAbs that exert a direct biological effect on tumor growth are useful to treat cancers that express 184P1E2. Mechanisms by which directly cytotoxic mAbs act include: inhibition of cell growth, modulation of cellular differentiation, modulation of tumor angiogenesis factor profiles, and the induction of apoptosis. The mechanism(s) by which a particular anti-184P1E2 mAb exerts an anti-tumor effect is evaluated using any number of in vitro assays that evaluate cell death such as ADCC, ADMMC, complement-mediated cell lysis, and so forth, as is generally known in the art.
In some patients, the use of murine or other non-human monoclonal antibodies, or human/mouse chimeric mAbs can induce moderate to strong immune responses against the non-human antibody. This can result in clearance of the antibody from circulation and reduced efficacy. In the most severe cases, such an immune response can lead to the extensive formation of immune complexes which, potentially, can cause renal failure. Accordingly, preferred monoclonal antibodies used in the therapeutic methods of the invention are those that are either fully human or humanized and that bind specifically to the target 184P1E2 antigen with high affinity but exhibit low or no antigenicity in the patient.
Therapeutic methods of the invention contemplate the administration of single anti-184P1E2 mAbs as well as combinations, or cocktails, of different mAbs. Such mAb cocktails can have certain advantages inasmuch as they contain mAbs that target different epitopes, exploit different effector mechanisms or combine directly cytotoxic mAbs with mAbs that rely on immune effector functionality. Such mAbs in combination can exhibit synergistic therapeutic effects. In addition, anti-184P1E2 mAbs can be administered concomitantly with other therapeutic modalities, including but not limited to various chemotherapeutic agents, androgen-blockers, immune modulators (e.g., IL-2, GM-CSF), surgery or radiation. The anti-184P1E2 mAbs are administered in their “naked” or unconjugated form, or can have a therapeutic agent(s) conjugated to them.
Anti-184P1E2 antibody formulations are administered via any route capable of delivering the antibodies to a tumor cell. Routes of administration include, but are not limited to, intravenous, intraperitoneal, intramuscular, intratumor, intradermal, and the like. Treatment generally involves repeated administration of the anti-184P1E2 antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25 mg/kg body weight. In general, doses in the range of 10-1000 mg mAb per week are effective and well tolerated.
Based on clinical experience with the HerceptinTM mAb in the treatment of metastatic breast cancer, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV of the anti-184P1E2 mAb preparation represents an acceptable dosing regimen. Preferably, the initial loading dose is administered as a 90 minute or longer infusion. The periodic maintenance dose is administered as a 30 minute or longer infusion, provided the initial dose was well tolerated. As appreciated by those of skill in the art, various factors can influence the ideal dose regimen in a particular case. Such factors include, for example, the binding affinity and half life of the Ab or mAbs used, the degree of 184P1E2 expression in the patient, the extent of circulating shed 184P1E2 antigen, the desired steady-state antibody concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient.
Optionally, patients should be evaluated for the levels of 184P1E2 in a given sample (e.g. the levels of circulating 184P1E2 antigen and/or 184P1E2 expressing cells) in order to assist in the determination of the most effective dosing regimen, etc. Such evaluations are also used for monitoring purposes throughout therapy, and are useful to gauge therapeutic success in combination with the evaluation of other parameters (for example, urine cytology and/or ImmunoCyt levels in bladder cancer therapy, or by analogy, serum PSA levels in prostate cancer therapy).
Anti-idiotypic anti-184P1E2 antibodies can also be used in anti-cancer therapy as a vaccine for inducing an immune response to cells expressing a 184P1E2-related protein. In particular, the generation of anti-idiotypic antibodies is well known in the art; this methodology can readily be adapted to generate anti-idiotypic anti-184P1E2 antibodies that mimic an epitope on a 184P1E2-related protein (see, for example, Wagner et al., 1997, Hybridoma 16: 33-40; Foon et al., 1995, J. Clin. Invest. 96:334-342; Herlyn et al., 1996, Cancer Immunol. Immunother. 43:65-76). Such an anti-idiotypic antibody can be used in cancer vaccine strategies.
X.C.) 184P1E2 as a Target for Cellular Immune Responses
Vaccines and methods of preparing vaccines that contain an immunogenically effective amount of one or more HLA-binding peptides as described herein are further embodiments of the invention. Furthermore, vaccines in accordance with the invention encompass compositions of one or more of the claimed peptides. A peptide can be present in a vaccine individually. Alternatively, the peptide can exist as a homopolymer comprising multiple copies of the same peptide, or as a heteropolymer of various peptides. Polymers have the advantage of increased immunological reaction and, where different peptide epitopes are used to make up the polymer, the additional ability to induce antibodies and/or CTLs that react with different antigenic determinants of the pathogenic organism or tumor-related peptide targeted for an immune response. The composition can be a naturally occurring region of an antigen or can be prepared, e.g., recombinantly or by chemical synthesis.
Carriers that can be used with vaccines of the invention are well known in the art, and include, e.g., thyroglobulin, albumins such as human serum albumin, tetanus toxoid, polyamino acids such as poly L-lysine, poly L-glutamic acid, influenza, hepatitis B virus core protein, and the like. The vaccines can contain a physiologically tolerable (i.e., acceptable) diluent such as water, or saline, preferably phosphate buffered saline. The vaccines also typically include an adjuvant. Adjuvants such as incomplete Freund's adjuvant, aluminum phosphate, aluminum hydroxide, or alum are examples of materials well known in the art. Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides of the invention to lipids, such as tripalmitoyl-S-glycerylcysteinlyseryl-serine (P3CSS). Moreover, an adjuvant such as a synthetic cytosine-phosphorothiolated-guanine-containing (CpG) oligonucleotides has been found to increase CTL responses 10- to 100-fold. (see, e.g. Davila and Celis, J. Immunol. 165:539-547 (2000))
Upon immunization with a peptide composition in accordance with the invention, via injection, aerosol, oral, transdermal, transmucosal, intrapleural, intrathecal, or other suitable routes, the immune system of the host responds to the vaccine by producing large amounts of CTLs and/or HTLs specific for the desired antigen. Consequently, the host becomes at least partially immune to later development of cells that express or overexpress 184P1E2 antigen, or derives at least some therapeutic benefit when the antigen was tumor-associated.
In some embodiments, it may be desirable to combine the class I peptide components with components that induce or facilitate neutralizing antibody and or helper T cell responses directed to the target antigen. A preferred embodiment of such a composition comprises class I and class II epitopes in accordance with the invention. An alternative embodiment of such a composition comprises a class I and/or class II epitope in accordance with the invention, along with a cross reactive HTL epitope such as PADRETM (Epimmune, San Diego, Calif.) molecule (described e.g., in U.S. Pat. No. 5,736,142).
A vaccine of the invention can also include antigen-presenting cells (APC), such as dendritic cells (DC), as a vehicle to present peptides of the invention. Vaccine compositions can be created in vitro, following dendritic cell mobilization and harvesting, whereby loading of dendritic cells occurs in vitro. For example, dendritic cells are transfected, e.g., with a minigene in accordance with the invention, or are pulsed with peptides. The dendritic cell can then be administered to a patient to elicit immune responses in vivo. Vaccine compositions, either DNA- or peptide-based, can also be administered in vivo in combination with dendritic cell mobilization whereby loading of dendritic cells occurs in vivo.
Preferably, the following principles are utilized when selecting an array of epitopes for inclusion in a polyepitopic composition for use in a vaccine, or for selecting discrete epitopes to be included in a vaccine and/or to be encoded by nucleic acids such as a minigene. It is preferred that each of the following principles be balanced in order to make the selection. The multiple epitopes to be incorporated in a given vaccine composition may be, but need not be, contiguous in sequence in the native antigen from which the epitopes are derived.
1.) Epitopes are selected which, upon administration, mimic immune responses that have been observed to be correlated with tumor clearance. For HLA Class I this includes 3-4 epitopes that come from at least one tumor associated antigen (TAA). For HLA Class II a similar rationale is employed; again 3-4 epitopes are selected from at least one TAA (see, e.g., Rosenberg et al., Science 278:1447-1450). Epitopes from one TAA may be used in combination with epitopes from one or more additional TAAs to produce a vaccine that targets tumors with varying expression patterns of frequently-expressed TAAs.
2.) Epitopes are selected that have the requisite binding affinity established to be correlated with immunogenicity: for HLA Class I an IC50 of 500 nM or less, often 200 nM or less; and for Class II an IC50 of 1000 nM or less.
3.) Sufficient supermotif bearing-peptides, or a sufficient array of allele-specific motif-bearing peptides, are selected to give broad population coverage. For example, it is preferable to have at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess the breadth, or redundancy of, population coverage.
4.) When selecting epitopes from cancer-related antigens it is often useful to select analogs because the patient may have developed tolerance to the native epitope.
5.) Of particular relevance are epitopes referred to as “nested epitopes.” Nested epitopes occur where at least two epitopes overlap in a given peptide sequence. A nested peptide sequence can comprise B cell, HLA class I and/or HLA class II epitopes. When providing nested epitopes, a general objective is to provide the greatest number of epitopes per sequence. Thus, an aspect is to avoid providing a peptide that is any longer than the amino terminus of the amino terminal epitope and the carboxyl terminus of the carboxyl terminal epitope in the peptide. When providing a multi-epitopic sequence, such as a sequence comprising nested epitopes, it is generally important to screen the sequence in order to insure that it does not have pathological or other deleterious biological properties.
6.) If a polyepitopic protein is created, or when creating a minigene, an objective is to generate the smallest peptide that encompasses the epitopes of interest. This principle is similar, if not the same as that employed when selecting a peptide comprising nested epitopes. However, with an artificial polyepitopic peptide, the size minimization objective is balanced against the need to integrate any spacer sequences between epitopes in the polyepitopic protein. Spacer amino acid residues can, for example, be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not present in the target antigen, and only created by the man-made juxtaposition of epitopes), or to facilitate cleavage between epitopes and thereby enhance epitope presentation. Junctional epitopes are generally to be avoided because the recipient may generate an immune response to that non-native epitope. Of particular concern is a junctional epitope that is a “dominant epitope.” A dominant epitope may lead to such a zealous response that immune responses to other epitopes are diminished or suppressed.
7.) Where the sequences of multiple variants of the same target protein are present, potential peptide epitopes can also be selected on the basis of their conservancy. For example, a criterion for conservancy may define that the entire sequence of an HLA class I binding peptide or the entire 9-mer core of a class II binding peptide be conserved in a designated percentage of the sequences evaluated for a specific protein antigen.
X.C.1. Minigene Vaccines
A number of different approaches are available which allow simultaneous delivery of multiple epitopes. Nucleic acids encoding the peptides of the invention are a particularly useful embodiment of the invention. Epitopes for inclusion in a minigene are preferably selected according to the guidelines set forth in the previous section. A preferred means of administering nucleic acids encoding the peptides of the invention uses minigene constructs encoding a peptide comprising one or multiple epitopes of the invention.
The use of multi-epitope minigenes is described below and in, Ishioka et al., J. Immunol. 162:3915-3925, 1999; An, L. and Whitton, J. L., J. Virol. 71:2292, 1997; Thomson, S. A. et al., J. Immunol. 157:822, 1996; Whitton, J. L. et al., J. Virol. 67:348, 1993; Hanke, R. et al., Vaccine 16:426, 1998. For example, a multi-epitope DNA plasmid encoding supermotif- and/or motif-bearing epitopes derived 184P1E2, the PADRE® universal helper T cell epitope or multiple HTL epitopes from 184P1E2 (see e.g., Tables V-XVIII and XXII to LI), and an endoplasmic reticulum-translocating signal sequence can be engineered. A vaccine may also comprise epitopes that are derived from other TAAs.
The immunogenicity of a multi-epitopic minigene can be confirmed in transgenic mice to evaluate the magnitude of CTL induction responses against the epitopes tested. Further, the immunogenicity of DNA-encoded epitopes in vivo can be correlated with the in vitro responses of specific CTL lines against target cells transfected with the DNA plasmid. Thus, these experiments can show that the minigene serves to both: 1.) generate a CTL response and 2.) that the induced CTLs recognized cells expressing the encoded epitopes.
For example, to create a DNA sequence encoding the selected epitopes (minigene) for expression in human cells, the amino acid sequences of the epitopes may be reverse translated. A human codon usage table can be used to guide the codon choice for each amino acid. These epitope-encoding DNA sequences may be directly adjoined, so that when translated, a continuous polypeptide sequence is created. To optimize expression and/or immunogenicity, additional elements can be incorporated into the minigene design. Examples of amino acid sequences that can be reverse translated and included in the minigene sequence include: HLA class I epitopes, HLA class II epitopes, antibody epitopes, a ubiquitination signal sequence, and/or an endoplasmic reticulum targeting signal. In addition, HLA presentation of CTL and HTL epitopes may be improved by including synthetic (e.g. poly-alanine) or naturally-occurring flanking sequences adjacent to the CTL or HTL epitopes; these larger peptides comprising the epitope(s) are within the scope of the invention.
The minigene sequence may be converted to DNA by assembling oligonucleotides that encode the plus and minus strands of the minigene. Overlapping oligonucleotides (30-100 bases long) may be synthesized, phosphorylated, purified and annealed under appropriate conditions using well known techniques. The ends of the oligonucleotides can be joined, for example, using T4 DNA ligase. This synthetic minigene, encoding the epitope polypeptide, can then be cloned into a desired expression vector.
Standard regulatory sequences well known to those of skill in the art are preferably included in the vector to ensure expression in the target cells. Several vector elements are desirable: a promoter with a down-stream cloning site for minigene insertion; a polyadenylation signal for efficient transcription termination; an E. coli origin of replication; and an E. coli selectable marker (e.g. ampicillin or kanamycin resistance). Numerous promoters can be used for this purpose, e.g., the human cytomegalovirus (hCMV) promoter. See, e.g., U.S. Pat. Nos. 5,580,859 and 5,589,466 for other suitable promoter sequences.
Additional vector modifications may be desired to optimize minigene expression and immunogenicity. In some cases, introns are required for efficient gene expression, and one or more synthetic or naturally-occurring introns could be incorporated into the transcribed region of the minigene. The inclusion of mRNA stabilization sequences and sequences for replication in mammalian cells may also be considered for increasing minigene expression.
Once an expression vector is selected, the minigene is cloned into the polylinker region downstream of the promoter. This plasmid is transformed into an appropriate E. coli strain, and DNA is prepared using standard techniques. The orientation and DNA sequence of the minigene, as well as all other elements included in the vector, are confirmed using restriction mapping and DNA sequence analysis. Bacterial cells harboring the correct plasmid can be stored as a master cell bank and a working cell bank.
In addition, immunostimulatory sequences (ISSs or CpGs) appear to play a role in the immunogenicity of DNA vaccines. These sequences may be included in the vector, outside the minigene coding sequence, if desired to enhance immunogenicity.
In some embodiments, a bi-cistronic expression vector which allows production of both the minigene-encoded epitopes and a second protein (included to enhance or decrease immunogenicity) can be used. Examples of proteins or polypeptides that could beneficially enhance the immune response if co-expressed include cytokines (e.g., IL-2, IL-12, GM-CSF), cytokine-inducing molecules (e.g., LeIF), costimulatory molecules, or for HTL responses, pan-DR binding proteins (PADRETM, Epimmune, San Diego, Calif.). Helper (HTL) epitopes can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes; this allows direction of the HTL epitopes to a cell compartment different than that of the CTL epitopes. If required, this could facilitate more efficient entry of HTL epitopes into the HLA class II pathway, thereby improving HTL induction. In contrast to HTL or CTL induction, specifically decreasing the immune response by co-expression of immunosuppressive molecules (e.g. TGF-β) may be beneficial in certain diseases.
Therapeutic quantities of plasmid DNA can be produced for example, by fermentation in E. coli, followed by purification. Aliquots from the working cell bank are used to inoculate growth medium, and grown to saturation in shaker flasks or a bioreactor according to well-known techniques. Plasmid DNA can be purified using standard bioseparation technologies such as solid phase anion-exchange resins supplied by QIAGEN, Inc. (Valencia, Calif.). If required, supercoiled DNA can be isolated from the open circular and linear forms using gel electrophoresis or other methods.
Purified plasmid DNA can be prepared for injection using a variety of formulations. The simplest of these is reconstitution of lyophilized DNA in sterile phosphate-buffer saline (PBS). This approach, known as “naked DNA,” is currently being used for intramuscular (IM) administration in clinical trials. To maximize the immunotherapeutic effects of minigene DNA vaccines, an alternative method for formulating purified plasmid DNA may be desirable. A variety of methods have been described, and new techniques may become available. Cationic lipids, glycolipids, and fusogenic liposomes can also be used in the formulation (see, e.g., as described by WO 93/24640; Mannino & Gould-Fogerite, BioTechniques 6(7): 682 (1988); U.S. Pat. No. 5,279,833; WO 91/06309; and Feigner, et al., Proc. Nat'l Acad. Sci. USA 84:7413 (1987). In addition, peptides and compounds referred to collectively as protective, interactive, non-condensing compounds (PINC) could also be complexed to purified plasmid DNA to influence variables such as stability, intramuscular dispersion, or trafficking to specific organs or cell types.
Target cell sensitization can be used as a functional assay for expression and HLA class I presentation of minigene-encoded CTL epitopes. For example, the plasmid DNA is introduced into a mammalian cell line that is suitable as a target for standard CTL chromium release assays. The transfection method used will be dependent on the final formulation. Electroporation can be used for “naked” DNA, whereas cationic lipids allow direct in vitro transfection. A plasmid expressing green fluorescent protein (GFP) can be co-transfected to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). These cells are then chromium-51 (51Cr) labeled and used as target cells for epitope-specific CTL lines; cytolysis, detected by 51Cr release, indicates both production of, and HLA presentation of, minigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated in an analogous manner using assays to assess HTL activity.
In vivo immunogenicity is a second approach for functional testing of minigene DNA formulations. Transgenic mice expressing appropriate human HLA proteins are immunized with the DNA product. The dose and route of administration are formulation dependent (e.g., IM for DNA in PBS, intraperitoneal (i.p.) for lipid-complexed DNA). Twenty-one days after immunization, splenocytes are harvested and restimulated for one week in the presence of peptides encoding each epitope being tested. Thereafter, for CTL effector cells, assays are conducted for cytolysis of peptide-loaded, 51Cr-labeled target cells using standard techniques. Lysis of target cells that were sensitized by HLA loaded with peptide epitopes, corresponding to minigene-encoded epitopes, demonstrates DNA vaccine function for in vivo induction of CTLs. Immunogenicity of HTL epitopes is confirmed in transgenic mice in an analogous manner.
Alternatively, the nucleic acids can be administered using ballistic delivery as described, for instance, in U.S. Pat. No. 5,204,253. Using this technique, particles comprised solely of DNA are administered. In a further alternative embodiment, DNA can be adhered to particles, such as gold particles.
Minigenes can also be delivered using other bacterial or viral delivery systems well known in the art, e.g., an expression construct encoding epitopes of the invention can be incorporated into a viral vector such as vaccinia.
X.C.2. Combinations of CTL Peptides with Helper Peptides
Vaccine compositions comprising CTL peptides of the invention can be modified, e.g., analoged, to provide desired attributes, such as improved serum half life, broadened population coverage or enhanced immunogenicity.
For instance, the ability of a peptide to induce CTL activity can be enhanced by linking the peptide to a sequence which contains at least one epitope that is capable of inducing a T helper cell response. Although a CTL peptide can be directly linked to a T helper peptide, often CTL epitope/HTL epitope conjugates are linked by a spacer molecule. The spacer is typically comprised of relatively small, neutral molecules, such as amino acids or amino acid mimetics, which are substantially uncharged under physiological conditions. The spacers are typically selected from, e.g., Ala, Gly, or other neutral spacers of nonpolar amino acids or neutral polar amino acids. It will be understood that the optionally present spacer need not be comprised of the same residues and thus may be a hetero- or homo-oligomer. When present, the spacer will usually be at least one or two residues, more usually three to six residues and sometimes 10 or more residues. The CTL peptide epitope can be linked to the T helper peptide epitope either directly or via a spacer either at the amino or carboxy terminus of the CTL peptide. The amino terminus of either the immunogenic peptide or the T helper peptide may be acylated.
In certain embodiments, the T helper peptide is one that is recognized by T helper cells present in a majority of a genetically diverse population. This can be accomplished by selecting peptides that bind to many, most, or all of the HLA class II molecules. Examples of such amino acid bind many HLA Class II molecules include sequences from antigens such as tetanus toxoid at positions 830-843 (QYIKANSKFIGITE; SEQ ID NO: 29), Plasmodium falciparum circumsporozoite (CS) protein at positions 378-398 (DIEKKIAKMEKASSVFNVVNS; SEQ ID NO: 30), and Streptococcus 18 kD protein at positions 116-131 (GAVDSILGGVATYGAA; SEQ ID NO: 31). Other examples include peptides bearing a DR 1-4-7 supermotif, or either of the DR3 motifs.
Alternatively, it is possible to prepare synthetic peptides capable of stimulating T helper lymphocytes, in a loosely HLA-restricted fashion, using amino acid sequences not found in nature (see, e.g., PCT publication WO 95/07707). These synthetic compounds called Pan-DR-binding epitopes (e.g., PADRETM, Epimmune, Inc., San Diego, Calif.) are designed to most preferably bind most HLA-DR (human HLA class II) molecules. For instance, a pan-DR-binding epitope peptide having the formula: aKXVAAWTLKAAa (SEQ ID NO: 32), where “X” is either cyclohexylalanine, phenylalanine, or tyrosine, and a is either D-alanine or L-alanine, has been found to bind to most HLA-DR alleles, and to stimulate the response of T helper lymphocytes from most individuals, regardless of their HLA type. An alternative of a pan-DR binding epitope comprises all “L” natural amino acids and can be provided in the form of nucleic acids that encode the epitope.
HTL peptide epitopes can also be modified to alter their biological properties. For example, they can be modified to include D-amino acids to increase their resistance to proteases and thus extend their serum half life, or they can be conjugated to other molecules such as lipids, proteins, carbohydrates, and the like to increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic acid chains at either the amino or carboxyl termini.
X.C.3. Combinations of CTL Peptides with T Cell Priming Agents
In some embodiments it may be desirable to include in the pharmaceutical compositions of the invention at least one component which primes B lymphocytes or T lymphocytes. Lipids have been identified as agents capable of priming CTL in vivo. For example, palmitic acid residues can be attached to the ε- and α-amino groups of a lysine residue and then linked, e.g., via one or more linking residues such as Gly, Gly-Gly-, Ser, Ser-Ser, or the like, to an immunogenic peptide. The lipidated peptide can then be administered either directly in a micelle or particle, incorporated into a liposome, or emulsified in an adjuvant, e.g., incomplete Freund's adjuvant. In a preferred embodiment, a particularly effective immunogenic composition comprises palmitic acid attached to ε- and α-amino groups of Lys, which is attached via linkage, e.g., Ser-Ser, to the amino terminus of the immunogenic peptide.
As another example of lipid priming of CTL responses, E. coli lipoproteins, such as tripalmitoyl-S-glycerylcysteinlyseryl-serine (P3CSS) can be used to prime virus specific CTL when covalently attached to an appropriate peptide (see, e.g., Deres, et al., Nature 342:561, 1989). Peptides of the invention can be coupled to P3CSS, for example, and the lipopeptide administered to an individual to specifically prime an immune response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed with P3CSS-conjugated epitopes, two such compositions can be combined to more effectively elicit both humoral and cell-mediated responses.
X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or HTL Peptides
An embodiment of a vaccine composition in accordance with the invention comprises ex vivo administration of a cocktail of epitope-bearing peptides to PBMC, or isolated DC therefrom, from the patient's blood. A pharmaceutical to facilitate harvesting of DC can be used, such as ProgenipoietinTM (Pharmacia-Monsanto, St. Louis, Mo.) or GM-CSF/IL-4. After pulsing the DC with peptides and prior to reinfusion into patients, the DC are washed to remove unbound peptides. In this embodiment, a vaccine comprises peptide-pulsed DCs which present the pulsed peptide epitopes complexed with HLA molecules on their surfaces.
The DC can be pulsed ex vivo with a cocktail of peptides, some of which stimulate CTL responses to 184P1E2. Optionally, a helper T cell (HTL) peptide, such as a natural or artificial loosely restricted HLA Class II peptide, can be included to facilitate the CTL response. Thus, a vaccine in accordance with the invention is used to treat a cancer which expresses or overexpresses 184P1E2.
X.D. Adoptive Immunotherapy
Antigenic 184P1E2-related peptides are used to elicit a CTL and/or HTL response ex vivo, as well. The resulting CTL or HTL cells, can be used to treat tumors in patients that do not respond to other conventional forms of therapy, or will not respond to a therapeutic vaccine peptide or nucleic acid in accordance with the invention. Ex vivo CTL or HTL responses to a particular antigen are induced by incubating in tissue culture the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of antigen-presenting cells (APC), such as dendritic cells, and the appropriate immunogenic peptide. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused back into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cell (e.g., a tumor cell). Transfected dendritic cells may also be used as antigen presenting cells.
X.E. Administration of Vaccines for Therapeutic or Prophylactic Purposes
Pharmaceutical and vaccine compositions of the invention are typically used to treat and/or prevent a cancer that expresses or overexpresses 184P1E2. In therapeutic applications, peptide and/or nucleic acid compositions are administered to a patient in an amount sufficient to elicit an effective B cell, CTL and/or HTL response to the antigen and to cure or at least partially arrest or slow symptoms and/or complications. An amount adequate to accomplish this is defined as “therapeutically effective dose.” Amounts effective for this use will depend on, e.g., the particular composition administered, the manner of administration, the stage and severity of the disease being treated, the weight and general state of health of the patient, and the judgment of the prescribing physician.
For pharmaceutical compositions, the immunogenic peptides of the invention, or DNA encoding them, are generally administered to an individual already bearing a tumor that expresses 184P1E2. The peptides or DNA encoding them can be administered individually or as fusions of one or more peptide sequences. Patients can be treated with the immunogenic peptides separately or in conjunction with other treatments, such as surgery, as appropriate.
For therapeutic use, administration should generally begin at the first diagnosis of 184P1E2-associated cancer. This is followed by boosting doses until at least symptoms are substantially abated and for a period thereafter. The embodiment of the vaccine composition (i.e., including, but not limited to embodiments such as peptide cocktails, polyepitopic polypeptides, minigenes, or TAA-specific CTLs or pulsed dendritic cells) delivered to the patient may vary according to the stage of the disease or the patient's health status. For example, in a patient with a tumor that expresses 184P1E2, a vaccine comprising 184P1E2-specific CTL may be more efficacious in killing tumor cells in patient with advanced disease than alternative embodiments.
It is generally important to provide an amount of the peptide epitope delivered by a mode of administration sufficient to effectively stimulate a cytotoxic T cell response; compositions which stimulate helper T cell responses can also be given in accordance with this embodiment of the invention.
The dosage for an initial therapeutic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1,000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. Boosting dosages of between about 1.0 μg to about 50,000 μg of peptide pursuant to a boosting regimen over weeks to months may be administered depending upon the patient's response and condition as determined by measuring the specific activity of CTL and HTL obtained from the patient's blood. Administration should continue until at least clinical symptoms or laboratory tests indicate that the neoplasia, has been eliminated or reduced and for a period thereafter. The dosages, routes of administration, and dose schedules are adjusted in accordance with methodologies known in the art.
In certain embodiments, the peptides and compositions of the present invention are employed in serious disease states, that is, life-threatening or potentially life threatening situations. In such cases, as a result of the minimal amounts of extraneous substances and the relative nontoxic nature of the peptides in preferred compositions of the invention, it is possible and may be felt desirable by the treating physician to administer substantial excesses of these peptide compositions relative to these stated dosage amounts.
The vaccine compositions of the invention can also be used purely as prophylactic agents. Generally the dosage for an initial prophylactic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. This is followed by boosting dosages of between about 1.0 μg to about 50,000 μg of peptide administered at defined intervals from about four weeks to six months after the initial administration of vaccine. The immunogenicity of the vaccine can be assessed by measuring the specific activity of CTL and HTL obtained from a sample of the patient's blood.
The pharmaceutical compositions for therapeutic treatment are intended for parenteral, topical, oral, nasal, intrathecal, or local (e.g. as a cream or topical ointment) administration. Preferably, the pharmaceutical compositions are administered parentally, e.g., intravenously, subcutaneously, intradermally, or intramuscularly. Thus, the invention provides compositions for parenteral administration which comprise a solution of the immunogenic peptides dissolved or suspended in an acceptable carrier, preferably an aqueous carrier.
A variety of aqueous carriers may be used, e.g., water, buffered water, 0.8% saline, 0.3% glycine, hyaluronic acid and the like. These compositions may be sterilized by conventional, well-known sterilization techniques, or may be sterile filtered. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile solution prior to administration.
The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservatives, and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.
The concentration of peptides of the invention in the pharmaceutical formulations can vary widely, i.e., from less than about 0.1%, usually at or at least about 2% to as much as 20% to 50% or more by weight, and will be selected primarily by fluid volumes, viscosities, etc., in accordance with the particular mode of administration selected.
A human unit dose form of a composition is typically included in a pharmaceutical composition that comprises a human unit dose of an acceptable carrier, in one embodiment an aqueous carrier, and is administered in a volume/quantity that is known by those of skill in the art to be used for administration of such compositions to humans (see, e.g., Remington's Pharmaceutical Sciences, 17th Edition, A. Gennaro, Editor, Mack Publishing Co., Easton, Pa., 1985). For example a peptide dose for initial immunization can be from about 1 to about 50,000 μg, generally 100-5,000 μg, for a 70 kg patient. For example, for nucleic acids an initial immunization may be performed using an expression vector in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose is then administered. The booster can be recombinant fowlpox virus administered at a dose of 5-107 to 5×109 pfu.
For antibodies, a treatment generally involves repeated administration of the anti-184P1E2 antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1 to about 10 mg/kg body weight. In general, doses in the range of 10-500 mg mAb per week are effective and well tolerated. Moreover, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV of the anti-184P1E2 mAb preparation represents an acceptable dosing regimen. As appreciated by those of skill in the art, various factors can influence the ideal dose in a particular case. Such factors include, for example, half life of a composition, the binding affinity of an Ab, the immunogenicity of a substance, the degree of 184P1E2 expression in the patient, the extent of circulating shed 184P1E2 antigen, the desired steady-state concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient. Non-limiting preferred human unit doses are, for example, 500 μg -1 mg, 1 mg -50 mg, 50 mg -100 mg, 100 mg -200 mg, 200 mg -300 mg, 400 mg -500 mg, 500 mg -600 mg, 600 mg -700 mg, 700 mg-800 mg, 800 mg -900 mg, 900 mg-1 g, or 1 mg -700 mg. In certain embodiments, the dose is in a range of 2-5 mg/kg body weight, e.g., with follow on weekly doses of 1-3 mg/kg; 0.5 mg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 mg/kg body weight followed, e.g., in two, three or four weeks by weekly doses; 0.5-10 mg/kg body weight, e.g., followed in two, three or four weeks by weekly doses; 225, 250, 275, 300, 325, 350, 375, 400 mg m2 of body area weekly; 1-600 mg m2 of body area weekly; 225-400 mg m2 of body area weekly; these does can be followed by weekly doses for 2, 3, 4, 5, 6, 7, 8, 9, 19, 11, 12 or more weeks.
In one embodiment, human unit dose forms of polynucleotides comprise a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art a therapeutic effect depends on a number of factors, including the sequence of the polynucleotide, molecular weight of the polynucleotide and route of administration. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. Generally, for a polynucleotide of about 20 bases, a dosage range may be selected from, for example, an independently selected lower limit such as about 0.1, 0.25, 0.5, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 mg/kg up to an independently selected upper limit, greater than the lower limit, of about 60, 80, 100, 200, 300, 400, 500, 750, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000 or 10,000 mg/kg. For example, a dose may be about any of the following: 0.1 to 100 mg/kg, 0.1 to 50 mg/kg, 0.1 to 25 mg/kg, 0.1 to 10 mg/kg, 1 to 500 mg/kg, 100 to 400 mg/kg, 200 to 300 mg/kg, 1 to 100 mg/kg, 100 to 200 mg/kg, 300 to 400 mg/kg, 400 to 500 mg/kg, 500 to 1000 mg/kg, 500 to 5000 mg/kg, or 500 to 10,000 mg/kg. Generally, parenteral routes of administration may require higher doses of polynucleotide compared to more direct application to the nucleotide to diseased tissue, as do polynucleotides of increasing length.
In one embodiment, human unit dose forms of T-cells comprise a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art, a therapeutic effect depends on a number of factors. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. A dose may be about 104 cells to about 106 cells, about 106 cells to about 108 cells, about 108 to about 1011 cells, or about 108 to about 5×1010 cells. A dose may also about 106 cells/m2 to about 1010 cells/m2, or about 106 cells/m2 to about 108 cells/m2.
Proteins(s) of the invention, and/or nucleic acids encoding the protein(s), can also be administered via liposomes, which may also serve to: 1) target the proteins(s) to a particular tissue, such as lymphoid tissue; 2) to target selectively to diseases cells; or, 3) to increase the half-life of the peptide composition. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. In these preparations, the peptide to be delivered is incorporated as part of a liposome, alone or in conjunction with a molecule which binds to a receptor prevalent among lymphoid cells, such as monoclonal antibodies which bind to the CD45 antigen, or with other therapeutic or immunogenic compositions. Thus, liposomes either filled or decorated with a desired peptide of the invention can be directed to the site of lymphoid cells, where the liposomes then deliver the peptide compositions. Liposomes for use in accordance with the invention are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of, e.g., liposome size, acid lability and stability of the liposomes in the blood stream. A variety of methods are available for preparing liposomes, as described in, e.g., Szoka, et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980), and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.
For targeting cells of the immune system, a ligand to be incorporated into the liposome can include, e.g., antibodies or fragments thereof specific for cell surface determinants of the desired immune system cells. A liposome suspension containing a peptide may be administered intravenously, locally, topically, etc. in a dose which varies according to, inter alia, the manner of administration, the peptide being delivered, and the stage of the disease being treated.
For solid compositions, conventional nontoxic solid carriers may be used which include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like. For oral administration, a pharmaceutically acceptable nontoxic composition is formed by incorporating any of the normally employed excipients, such as those carriers previously listed, and generally 10-95% of active ingredient, that is, one or more peptides of the invention, and more preferably at a concentration of 25%-75%.
For aerosol administration, immunogenic peptides are preferably supplied in finely divided form along with a surfactant and propellant. Typical percentages of peptides are about 0.01%-20% by weight, preferably about 1%-10%. The surfactant must, of course, be nontoxic, and preferably soluble in the propellant. Representative of such agents are the esters or partial esters of fatty acids containing from about 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, linolenic, olesteric and oleic acids with an aliphatic polyhydric alcohol or its cyclic anhydride. Mixed esters, such as mixed or natural glycerides may be employed. The surfactant may constitute about 0.1%-20% by weight of the composition, preferably about 0.25-5%. The balance of the composition is ordinarily propellant. A carrier can also be included, as desired, as with, e.g., lecithin for intranasal delivery.
XI.) Diagnostic and Prognostic Embodiments of 184P1E2
As disclosed herein, 184P1E2 polynucleotides, polypeptides, reactive cytotoxic T cells (CTL), reactive helper T cells (HTL) and anti-polypeptide antibodies are used in well known diagnostic, prognostic and therapeutic assays that examine conditions associated with dysregulated cell growth such as cancer, in particular the cancers listed in Table I (see, e.g., both its specific pattern of tissue expression as well as its overexpression in certain cancers as described for example in the Example entitled “Expression analysis of 184P1E2 in normal tissues, and patient specimens”).
184P1E2 can be analogized to a prostate associated antigen PSA, the archetypal marker that has been used by medical practitioners for years to identify and monitor the presence of prostate cancer (see, e.g., Merrill et al., J. Urol. 163(2): 503-5120 (2000); Polascik et al., J. Urol. Aug; 162(2):293-306 (1999) and Fortier et al., J. Nat. Cancer Inst. 91(19): 1635-1640 (1999)). A variety of other diagnostic markers are also used in similar contexts including p53 and K-ras (see, e.g., Tulchinsky et al., Int J Mol Med 1999 July 4(1):99-102 and Minimoto et al., Cancer Detect Prev 2000; 24(1):1-12). Therefore, this disclosure of 184P1E2 polynucleotides and polypeptides (as well as 184P1E2 polynucleotide probes and anti-184P1E2 antibodies used to identify the presence of these molecules) and their properties allows skilled artisans to utilize these molecules in methods that are analogous to those used, for example, in a variety of diagnostic assays directed to examining conditions associated with cancer.
Typical embodiments of diagnostic methods which utilize the 184P1E2 polynucleotides, polypeptides, reactive T cells and antibodies are analogous to those methods from well-established diagnostic assays which employ, e.g., PSA polynucleotides, polypeptides, reactive T cells and antibodies. For example, just as PSA polynucleotides are used as probes (for example in Northern analysis, see, e.g., Sharief et al., Biochem. Mol. Biol. Int. 33(3):567-74 (1994)) and primers (for example in PCR analysis, see, e.g., Okegawa et al., J. Urol. 163(4): 1189-1190 (2000)) to observe the presence and/or the level of PSA mRNAs in methods of monitoring PSA overexpression or the metastasis of prostate cancers, the 184P1E2 polynucleotides described herein can be utilized in the same way to detect 184P1E2 overexpression or the metastasis of prostate and other cancers expressing this gene. Alternatively, just as PSA polypeptides are used to generate antibodies specific for PSA which can then be used to observe the presence and/or the level of PSA proteins in methods to monitor PSA protein overexpression (see, e.g., Stephan et al., Urology 55(4):560-3 (2000)) or the metastasis of prostate cells (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3):233-7 (1996)), the 184P1E2 polypeptides described herein can be utilized to generate antibodies for use in detecting 184P1E2 overexpression or the metastasis of prostate cells and cells of other cancers expressing this gene.
Specifically, because metastases involves the movement of cancer cells from an organ of origin (such as the lung or prostate gland etc.) to a different area of the body (such as a lymph node), assays which examine a biological sample for the presence of cells expressing 184P1E2 polynucleotides and/or polypeptides can be used to provide evidence of metastasis. For example, when a biological sample from tissue that does not normally contain 184P1E2-expressing cells (lymph node) is found to contain 184P1E2-expressing cells such as the 184P1E2 expression seen in LAPC4 and LAPC9, xenografts isolated from lymph node and bone metastasis, respectively, this finding is indicative of metastasis.
Alternatively 184P1E2 polynucleotides and/or polypeptides can be used to provide evidence of cancer, for example, when cells in a biological sample that do not normally express 184P1E2 or express 184P1E2 at a different level are found to express 184P1E2 or have an increased expression of 184P1E2 (see, e.g., the 184P1E2 expression in the cancers listed in Table I and in patient samples etc. shown in the accompanying Figures). In such assays, artisans may further wish to generate supplementary evidence of metastasis by testing the biological sample for the presence of a second tissue restricted marker (in addition to 184P1E2) such as PSA, PSCA etc. (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3): 233-237 (1996)).
Just as PSA polynucleotide fragments and polynucleotide variants are employed by skilled artisans for use in methods of monitoring PSA, 184P1E2 polynucleotide fragments and polynucleotide variants are used in an analogous manner. In particular, typical PSA polynucleotides used in methods of monitoring PSA are probes or primers which consist of fragments of the PSA cDNA sequence. Illustrating this, primers used to PCR amplify a PSA polynucleotide must include less than the whole PSA sequence to function in the polymerase chain reaction. In the context of such PCR reactions, skilled artisans generally create a variety of different polynucleotide fragments that can be used as primers in order to amplify different portions of a polynucleotide of interest or to optimize amplification reactions (see, e.g., Caetano-Anolles, G. Biotechniques 25(3): 472-476, 478-480 (1998); Robertson et al., Methods Mol. Biol. 98:121-154 (1998)). An additional illustration of the use of such fragments is provided in the Example entitled “Expression analysis of 184P1E2 in normal tissues, and patient specimens,” where a 184P1E2 polynucleotide fragment is used as a probe to show the expression of 184P1E2 RNAs in cancer cells. In addition, variant polynucleotide sequences are typically used as primers and probes for the corresponding mRNAs in PCR and Northern analyses (see, e.g., Sawai et al., Fetal Diagn. Ther. 1996 November-December 11(6):407-13 and Current Protocols In Molecular Biology, Volume 2, Unit 2, Frederick M. Ausubel et al. eds., 1995)). Polynucleotide fragments and variants are useful in this context where they are capable of binding to a target polynucleotide sequence (e.g., a 184P1E2 polynucleotide shown in FIG. 2 or variant thereof) under conditions of high stringency.
Furthermore, PSA polypeptides which contain an epitope that can be recognized by an antibody or T cell that specifically binds to that epitope are used in methods of monitoring PSA. 184P1E2 polypeptide fragments and polypeptide analogs or variants can also be used in an analogous manner. This practice of using polypeptide fragments or polypeptide variants to generate antibodies (such as anti-PSA antibodies or T cells) is typical in the art with a wide variety of systems such as fusion proteins being used by practitioners (see, e.g., Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubel et al. eds., 1995). In this context, each epitope(s) functions to provide the architecture with which an antibody or T cell is reactive. Typically, skilled artisans create a variety of different polypeptide fragments that can be used in order to generate immune responses specific for different portions of a polypeptide of interest (see, e.g., U.S. Pat. No. 5,840,501 and U.S. Pat. No. 5,939,533). For example it may be preferable to utilize a polypeptide comprising one of the 184P1E2 biological motifs discussed herein or a motif-bearing subsequence which is readily identified by one of skill in the art based on motifs available in the art. Polypeptide fragments, variants or analogs are typically useful in this context as long as they comprise an epitope capable of generating an antibody or T cell specific for a target polypeptide sequence (e.g. a 184P1E2 polypeptide shown in FIG. 3).
As shown herein, the 184P1E2 polynucleotides and polypeptides (as well as the 184P1E2 polynucleotide probes and anti-184P1E2 antibodies or T cells used to identify the presence of these molecules) exhibit specific properties that make them useful in diagnosing cancers such as those listed in Table I. Diagnostic assays that measure the presence of 184P1E2 gene products, in order to evaluate the presence or onset of a disease condition described herein, such as prostate cancer, are used to identify patients for preventive measures or further monitoring, as has been done so successfully with PSA. Moreover, these materials satisfy a need in the art for molecules having similar or complementary characteristics to PSA in situations where, for example, a definite diagnosis of metastasis of prostatic origin cannot be made on the basis of a test for PSA alone (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3): 233-237 (1996)), and consequently, materials such as 184P1E2 polynucleotides and polypeptides (as well as the 184P1E2 polynucleotide probes and anti-184P1E2 antibodies used to identify the presence of these molecules) need to be employed to confirm a metastases of prostatic origin.
Finally, in addition to their use in diagnostic assays, the 184P1E2 polynucleotides disclosed herein have a number of other utilities such as their use in the identification of oncogenetic associated chromosomal abnormalities in the chromosomal region to which the 184P1E2 gene maps (see the Example entitled “Chromosomal Mapping of 184P1E2” below). Moreover, in addition to their use in diagnostic assays, the 184P1E2-related proteins and polynucleotides disclosed herein have other utilities such as their use in the forensic analysis of tissues of unknown origin (see, e.g., Takahama K Forensic Sci Int 1996 Jun. 28; 80(1-2): 63-9).
Additionally, 184P1E2-related proteins or polynucleotides of the invention can be used to treat a pathologic condition characterized by the over-expression of 184P1E2. For example, the amino acid or nucleic acid sequence of FIG. 2 or FIG. 3, or fragments of either, can be used to generate an immune response to a 184P1E2 antigen. Antibodies or other molecules that react with 184P1E2 can be used to modulate the function of this molecule, and thereby provide a therapeutic benefit.
XII.) Inhibition of 184P1E2 Protein Function
The invention includes various methods and compositions for inhibiting the binding of 184P1E2 to its binding partner or its association with other protein(s) as well as methods for inhibiting 184P1E2 function.
XII.A.) Inhibition of 184P1E2 with Intracellular Antibodies
In one approach, a recombinant vector that encodes single chain antibodies that specifically bind to 184P1E2 are introduced into 184P1E2 expressing cells via gene transfer technologies. Accordingly, the encoded single chain anti-184P1E2 antibody is expressed intracellularly, binds to 184P1E2 protein, and thereby inhibits its function. Methods for engineering such intracellular single chain antibodies are well known. Such intracellular antibodies, also known as “intrabodies”, are specifically targeted to a particular compartment within the cell, providing control over where the inhibitory activity of the treatment is focused. This technology has been successfully applied in the art (for review, see Richardson and Marasco, 1995, TIBTECH vol. 13). Intrabodies have been shown to virtually eliminate the expression of otherwise abundant cell surface receptors (see, e.g., Richardson et al., 1995, Proc. Natl. Acad. Sci. USA 92: 3137-3141; Beerli et al., 1994, J. Biol. Chem. 289: 23931-23936; Deshane et al., 1994, Gene Ther. 1: 332-337).
Single chain antibodies comprise the variable domains of the heavy and light chain joined by a flexible linker polypeptide, and are expressed as a single polypeptide. Optionally, single chain antibodies are expressed as a single chain variable region fragment joined to the light chain constant region. Well-known intracellular trafficking signals are engineered into recombinant polynucleotide vectors encoding such single chain antibodies in order to precisely target the intrabody to the desired intracellular compartment. For example, intrabodies targeted to the endoplasmic reticulum (ER) are engineered to incorporate a leader peptide and, optionally, a C-terminal ER retention signal, such as the KDEL amino acid motif. Intrabodies intended to exert activity in the nucleus are engineered to include a nuclear localization signal. Lipid moieties are joined to intrabodies in order to tether the intrabody to the cytosolic side of the plasma membrane. Intrabodies can also be targeted to exert function in the cytosol. For example, cytosolic intrabodies are used to sequester factors within the cytosol, thereby preventing them from being transported to their natural cellular destination.
In one embodiment, intrabodies are used to capture 184P1E2 in the nucleus, thereby preventing its activity within the nucleus. Nuclear targeting signals are engineered into such 184P1E2 intrabodies in order to achieve the desired targeting. Such 184P1E2 intrabodies are designed to bind specifically to a particular 184P1E2 domain. In another embodiment, cytosolic intrabodies that specifically bind to a 184P1E2 protein are used to prevent 184P1E2 from gaining access to the nucleus, thereby preventing it from exerting any biological activity within the nucleus (e.g., preventing 184P1E2 from forming transcription complexes with other factors).
In order to specifically direct the expression of such intrabodies to particular cells, the transcription of the intrabody is placed under the regulatory control of an appropriate tumor-specific promoter and/or enhancer. In order to target intrabody expression specifically to prostate, for example, the PSA promoter and/or promoter/enhancer can be utilized (See, for example, U.S. Pat. No. 5,919,652 issued 6 Jul. 1999).
XII.B.) Inhibition of 184P1E2 with Recombinant Proteins
In another approach, recombinant molecules bind to 184P1E2 and thereby inhibit 184P1E2 function. For example, these recombinant molecules prevent or inhibit 184P1E2 from accessing/binding to its binding partner(s) or associating with other protein(s). Such recombinant molecules can, for example, contain the reactive part(s) of a 184P1E2 specific antibody molecule. In a particular embodiment, the 184P1E2 binding domain of a 184P1E2 binding partner is engineered into a dimeric fusion protein, whereby the fusion protein comprises two 184P1E2 ligand binding domains linked to the Fc portion of a human IgG, such as human IgG1. Such IgG portion can contain, for example, the CH2 and CH3 domains and the hinge region, but not the CH1 domain. Such dimeric fusion proteins are administered in soluble form to patients suffering from a cancer associated with the expression of 184P1E2, whereby the dimeric fusion protein specifically binds to 184P1E2 and blocks 184P1E2 interaction with a binding partner. Such dimeric fusion proteins are further combined into multimeric proteins using known antibody linking technologies.
XII.C.) Inhibition of 184P1E2 Transcription or Translation
The present invention also comprises various methods and compositions for inhibiting the transcription of the 184P1E2 gene. Similarly, the invention also provides methods and compositions for inhibiting the translation of 184P1E2 mRNA into protein.
In one approach, a method of inhibiting the transcription of the 184P1E2 gene comprises contacting the 184P1E2 gene with a 184P1E2 antisense polynucleotide. In another approach, a method of inhibiting 184P1E2 mRNA translation comprises contacting a 184P1E2 mRNA with an antisense polynucleotide. In another approach, a 184P1E2 specific ribozyme is used to cleave a 184P1E2 message, thereby inhibiting translation. Such antisense and ribozyme based methods can also be directed to the regulatory regions of the 184P1E2 gene, such as 184P1E2 promoter and/or enhancer elements. Similarly, proteins capable of inhibiting a 184P1E2 gene transcription factor are used to inhibit 184P1E2 mRNA transcription. The various polynucleotides and compositions useful in the aforementioned methods have been described above. The use of antisense and ribozyme molecules to inhibit transcription and translation is well known in the art.
Other factors that inhibit the transcription of 184P1E2 by interfering with 184P1E2 transcriptional activation are also useful to treat cancers expressing 184P1E2. Similarly, factors that interfere with 184P1E2 processing are useful to treat cancers that express 184P1E2. Cancer treatment methods utilizing such factors are also within the scope of the invention.
XII.D.) General Considerations for Therapeutic Strategies
Gene transfer and gene therapy technologies can be used to deliver therapeutic polynucleotide molecules to tumor cells synthesizing 184P1E2 (i.e., antisense, ribozyme, polynucleotides encoding intrabodies and other 184P1E2 inhibitory molecules). A number of gene therapy approaches are known in the art. Recombinant vectors encoding 184P1E2 antisense polynucleotides, ribozymes, factors capable of interfering with 184P1E2 transcription, and so forth, can be delivered to target tumor cells using such gene therapy approaches.
The above therapeutic approaches can be combined with any one of a wide variety of surgical, chemotherapy or radiation therapy regimens. The therapeutic approaches of the invention can enable the use of reduced dosages of chemotherapy (or other therapies) and/or less frequent administration, an advantage for all patients and particularly for those that do not tolerate the toxicity of the chemotherapeutic agent well.
The anti-tumor activity of a particular composition (e.g., antisense, ribozyme, intrabody), or a combination of such compositions, can be evaluated using various in vitro and in vivo assay systems. In vitro assays that evaluate therapeutic activity include cell growth assays, soft agar assays and other assays indicative of tumor promoting activity, binding assays capable of determining the extent to which a therapeutic composition will inhibit the binding of 184P1E2 to a binding partner, etc.
In vivo, the effect of a 184P1E2 therapeutic composition can be evaluated in a suitable animal model. For example, xenogenic prostate cancer models can be used, wherein human prostate cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et al., 1997, Nature Medicine 3: 402-408). For example, PCT Patent Application WO98/16628 and U.S. Pat. No. 6,107,540 describe various xenograft models of human prostate cancer capable of recapitulating the development of primary tumors, micrometastasis, and the formation of osteoblastic metastases characteristic of late stage disease. Efficacy can be predicted using assays that measure inhibition of tumor formation, tumor regression or metastasis, and the like.
In vivo assays that evaluate the promotion of apoptosis are useful in evaluating therapeutic compositions.
In one embodiment, xenografts from tumor bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in the tumors of the treated mice provides an indication of the therapeutic efficacy of the composition.
The therapeutic compositions used in the practice of the foregoing methods can be formulated into pharmaceutical compositions comprising a carrier suitable for the desired delivery method. Suitable carriers include any material that when combined with the therapeutic composition retains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient's immune system. Examples include, but are not limited to, any of a number of standard pharmaceutical carriers such as sterile phosphate buffered saline solutions, bacteriostatic water, and the like (see, generally, Remington's Pharmaceutical Sciences 16th Edition, A. Osal., Ed., 1980).
Therapeutic formulations can be solubilized and administered via any route capable of delivering the therapeutic composition to the tumor site. Potentially effective routes of administration include, but are not limited to, intravenous, parenteral, intraperitoneal, intramuscular, intratumor, intradermal, intraorgan, orthotopic, and the like. A preferred formulation for intravenous injection comprises the therapeutic composition in a solution of preserved bacteriostatic water, sterile unpreserved water, and/or diluted in polyvinylchloride or polyethylene bags containing 0.9% sterile Sodium Chloride for Injection, USP. Therapeutic protein preparations can be lyophilized and stored as sterile powders, preferably under vacuum, and then reconstituted in bacteriostatic water (containing for example, benzyl alcohol preservative) or in sterile water prior to injection.
Dosages and administration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated in the art.
XIII.) Kits
For use in the diagnostic and therapeutic applications described herein, kits are also within the scope of the invention. Such kits can comprise a carrier, package or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in the method. For example, the container(s) can comprise a probe that is or can be detectably labeled. Such probe can be an antibody or polynucleotide specific for a 184P1E2-related protein or a 184P1E2 gene or message, respectively. Where the method utilizes nucleic acid hybridization to detect the target nucleic acid, the kit can also have containers containing nucleotide(s) for amplification of the target nucleic acid sequence and/or a container comprising a reporter-means, such as a biotin-binding protein, such as avidin or streptavidin, bound to a reporter molecule, such as an enzymatic, florescent, or radioisotope label. The kit can include all or part of the amino acid sequence of FIG. 2 or FIG. 3 or analogs thereof, or a nucleic acid molecules that encodes such amino acid sequences.
The kit of the invention will typically comprise the container described above and one or more other containers comprising materials desirable from a commercial and user standpoint, including buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.
A label can be present on the container to indicate that the composition is used for a specific therapy or non-therapeutic application, and can also indicate directions for either in vivo or in vitro use, such as those described above. Directions and or other information can also be included on an insert which is included with the kit.
EXAMPLES
Various aspects of the invention are further described and illustrated by way of the several examples that follow, none of which are intended to limit the scope of the invention.
Example 1
SSH-Generated Isolation of a cDNA Fragment of the 184P1E2 Gene
To isolate genes that are over-expressed in bladder cancer, Suppression Subtractive Hybridization (SSH) procedure using cDNA derived from bladder cancer tissues, including invasive transitional cell carcinoma. The 184P1E2 SSH cDNA sequence was derived from a bladder cancer pool minus cDNAs derived normal bladder in addition to a pool of 9 normal tissues. The 184P1E2 cDNA was identified as highly expressed in the bladder cancer tissue pool, with no expression detected in normal tissues.
The SSH DNA sequence of 132 bp (FIG. 1) showed homology to peptidylarginine deiminase type III (AB026831) (FIG. 4A). 184P1E2 v.1 of 3183 bp was cloned from bladder cancer cDNA library, revealing an ORF of 664 amino acids (FIG. 2 and FIG. 3). The 184P1E2 v.1 protein is the same as the GenBank protein AB026831 with one amino acid difference at position 480 (FIG. 4B). Other variants of 184P1E2 were also identified, and these are listed in FIGS. 2 and 3. 184P1E2 v.3 is 100% identical to peptidylarginine deiminase type III Genbank protein AB026831 (see FIG. 4B and Table LIII).
Materials and Methods
Human Tissues:
The patient cancer and normal tissues were purchased from different sources such as the NDRI (Philadelphia, Pa.). mRNA for some normal tissues were purchased from Clontech, Palo Alto, Calif.
RNA Isolation:
Tissues were homogenized in Trizol reagent (Life Technologies, Gibco BRL) using 10 ml/g tissue isolate total RNA. Poly A RNA was purified from total RNA using Qiagen's Oligotex mRNA Mini and Midi kits. Total and mRNA were quantified by spectrophotometric analysis (O.D. 260/280 nm) and analyzed by gel electrophoresis.
Oligonucleotides:
The following HPLC purified oligonucleotides were used.








DPNCDN (cDNA synthesis primer):









(SEQ ID NO: 33)









5′TTTTGATCAAGCTT303′



 


Adaptor 1:








(SEQ ID NO: 34)









5′CTAATACGACTCACTATAGGGCTCGAGCGGCCGCCCGGGCAG3′



 








(SEQ ID NO: 35)









3′GGCCCGTCCTAG5′



 


Adaptor 2:








(SEQ ID NO: 36)









5′GTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAG3′



 








(SEQ ID NO: 37)









3′CGGCTCCTAG5′



 


PCR primer 1:








(SEQ ID NO: 38)









5′CTAATACGACTCACTATAGGGC3′



 


Nested primer (NP)1:








(SEQ ID NO: 39)









5′TCGAGCGGCCGCCCGGGCAGGA3′



 


Nested primer (NP)2:








(SEQ ID NO: 40)









5′AGCGTGGTCGCGGCCGAGGA3′







Suppression Subtractive Hybridization:
Suppression Subtractive Hybridization (SSH) was used to identify cDNAs corresponding to genes that may be differentially expressed in bladder cancer. The SSH reaction utilized cDNA from bladder cancer and normal tissues.
The gene 184P1E2 sequence was derived from a bladder cancer pool minus normal bladder cDNA subtraction. The SSH DNA sequence (FIG. 1) was identified.
The cDNA derived from of pool of normal tissues was used as the source of the “driver” cDNA, while the cDNA from a pool of bladder cancer tissues was used as the source of the “tester” cDNA. Double stranded cDNAs corresponding to tester and driver cDNAs were synthesized from 2 μg of poly(A)+ RNA isolated from the relevant xenograft tissue, as described above, using CLONTECH's PCR-Select cDNA Subtraction Kit and 1 ng of oligonucleotide DPNCDN as primer. First- and second-strand synthesis were carried out as described in the Kit's user manual protocol (CLONTECH Protocol No. PT1117-1, Catalog No. K1804-1). The resulting cDNA was digested with Dpn II for 3 hrs at 37° C. Digested cDNA was extracted with phenol/chloroform (1:1) and ethanol precipitated.
Driver cDNA was generated by combining in a 1:1 ratio Dpn II digested cDNA from the relevant tissue source (see above) with a mix of digested cDNAs derived from the nine normal tissues: stomach, skeletal muscle, lung, brain, liver, kidney, pancreas, small intestine, and heart.
Tester cDNA was generated by diluting 1 μl of Dpn II digested cDNA from the relevant tissue source (see above) (400 ng) in 5 μl of water. The diluted cDNA (2 μl, 160 ng) was then ligated to 2 μl of Adaptor 1 and Adaptor 2 (10 μM), in separate ligation reactions, in a total volume of 10111 at 16° C. overnight, using 400 u of T4 DNA ligase (CLONTECH). Ligation was terminated with 1 μl of 0.2 M EDTA and heating at 72° C. for 5 min.
The first hybridization was performed by adding 1.5 μl (600 ng) of driver cDNA to each of two tubes containing 1.5 μl (20 ng) Adaptor 1- and Adaptor 2-ligated tester cDNA. In a final volume of 4 μl, the samples were overlaid with mineral oil, denatured in an MJ Research thermal cycler at 98° C. for 1.5 minutes, and then were allowed to hybridize for 8 hrs at 68° C. The two hybridizations were then mixed together with an additional 1 μl of fresh denatured driver cDNA and were allowed to hybridize overnight at 68° C. The second hybridization was then diluted in 2001 of 20 mM Hepes, pH 8.3, 50 mM NaCl, 0.2 mM EDTA, heated at 70° C. for 7 min. and stored at −20° C.
PCR Amplification, Cloning and Sequencing of Gene Fragments Generated from SSH:
To amplify gene fragments resulting from SSH reactions, two PCR amplifications were performed. In the primary PCR reaction 1 μl of the diluted final hybridization mix was added to 1 μl of PCR primer 1 (10 μM), 0.5 μl dNTP mix (10 μM), 2.5 μl 10× reaction buffer (CLONTECH) and 0.5 μl 50× Advantage cDNA polymerase Mix (CLONTECH) in a final volume of 25 μl. PCR 1 was conducted using the following conditions: 75° C. for 5 min., 94° C. for 25 sec., then 27 cycles of 94° C. for 10 sec, 66° C. for 30 sec, 72° C. for 1.5 min. Five separate primary PCR reactions were performed for each experiment. The products were pooled and diluted 1:10 with water. For the secondary PCR reaction, 1 μl from the pooled and diluted primary PCR reaction was added to the same reaction mix as used for PCR 1, except that primers NP1 and NP2 (10 μM) were used instead of PCR primer 1. PCR 2 was performed using 10-12 cycles of 94° C. for 10 sec, 68° C. for 30 sec, and 72° C. for 1.5 minutes. The PCR products were analyzed using 2% agarose gel electrophoresis.
The PCR products were inserted into pCR2.1 using the T/A vector cloning kit (Invitrogen). Transformed E. coli were subjected to blue/white and ampicillin selection. White colonies were picked and arrayed into 96 well plates and were grown in liquid culture overnight. To identify inserts, PCR amplification was performed on 1 ul of bacterial culture using the conditions of PCR1 and NP1 and NP2 as primers. PCR products were analyzed using 2% agarose gel electrophoresis.
Bacterial clones were stored in 20% glycerol in a 96 well format. Plasmid DNA was prepared, sequenced, and subjected to nucleic acid homology searches of the GenBank, dBest, and NCI-CGAP databases.
RT-PCR Expression Analysis:
First strand cDNAs can be generated from 1 μg of mRNA with oligo (dT)12-18 priming using the Gibco-BRL Superscript Preamplification system. The manufacturer's protocol was used which included an incubation for 50 min at 42° C. with reverse transcriptase followed by RNAse H treatment at 37° C. for 20 min. After completing the reaction, the volume can be increased to 200 μl with water prior to normalization. First strand cDNAs from 16 different normal human tissues can be obtained from Clontech.
Normalization of the first strand cDNAs from multiple tissues was performed by using the primers 5′atatcgccgcgctcgtcgtcgacaa3′ (SEQ ID NO: 41) and 5′agccacacgcagctcattgtagaagg 3′ (SEQ ID NO: 42) to amplify β-actin. First strand cDNA (5 μl) were amplified in a total volume of 50 μl containing 0.4 μM primers, 0.2 μM each dNTPs, 1×PCR buffer (Clontech, 10 mM Tris-HCL, 1.5 mM MgCl2, 50 mM KCl, pH8.3) and 1× Klentaq DNA polymerase (Clontech). Five μl of the PCR reaction can be removed at 18, 20, and 22 cycles and used for agarose gel electrophoresis. PCR was performed using an MJ Research thermal cycler under the following conditions: Initial denaturation can be at 94° C. for 15 sec, followed by a 18, 20, and 22 cycles of 94° C. for 15, 65° C. for 2 min, 72° C. for 5 sec. A final extension at 72° C. was carried out for 2 min. After agarose gel electrophoresis, the band intensities of the 283 b.p. β-actin bands from multiple tissues were compared by visual inspection. Dilution factors for the first strand cDNAs were calculated to result in equal β-actin band intensities in all tissues after 22 cycles of PCR. Three rounds of normalization can be required to achieve equal band intensities in all tissues after 22 cycles of PCR.
To determine expression levels of the 184P1E2 gene, 5 μl of normalized first strand cDNA were analyzed by PCR using 26, and 30 cycles of amplification. Semi-quantitative expression analysis can be achieved by comparing the PCR products at cycle numbers that give light band intensities. The primers used for RT-PCR were designed using the 184P1E2 SSH sequence and are listed below:









184P1E2.1




5′- AGTGACATGGAAGGAGATGAGTCC -3′
(SEQ ID NO: 43)


 


184P1E2.2


5′- ATACCTCCAGCTATGATGCCAAAC -3′
(SEQ ID NO: 44)






A typical RT-PCR expression analysis is shown in FIG. 14. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P1E2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 184P1E2 in bladder cancer pool. Expression of 184P1E2 is also detected in kidney cancer pool, lung cancer pool and cancer metastasis pool, but not in vital pool 1 and vital pool 2.
Example 2
Full Length Cloning of 184P1E2
The 184P1E2 SSH cDNA sequence was derived from a bladder cancer pool minus normal bladder cDNA subtraction. The SSH cDNA sequence (FIG. 1) was designated 184P1E2.
The SSH DNA sequence of 132 bp (FIG. 1) showed homology to peptidylarginine deiminase type III (AB026831) (FIG. 4A). 184P1E2 v.1 of 3183 bp was cloned from bladder cancer cDNA library, revealing an ORF of 664 amino acids (FIG. 2 and FIG. 3). The 184P1E2 v.1 protein is the same as the GenBank protein AB026831 with one amino acid difference at position 480 (FIG. 4B). Other variants of 184P1E2 were also identified, and these are listed in FIGS. 2 and 3. 184P1E2 v.3 is 100% identical to peptidylarginine deiminase type III Genbank protein AB026831 (see FIG. 4B and Table LIII).
184P1E2 v.1 is 87% identical and 93% homologous to the mouse peptidylarginine deiminase type III protein. The amino acid alignment of these two proteins is shown in FIG. 4C.
Example 3
Chromosomal Mapping of 184P1E2
Chromosomal localization can implicate genes in disease pathogenesis. Several chromosome mapping approaches are available including fluorescent in situ hybridization (FISH), human/hamster radiation hybrid (RH) panels (Walter et al., 1994; Nature Genetics 7:22; Research Genetics, Huntsville Ala.), human-rodent somatic cell hybrid panels such as is available from the Coriell Institute (Camden, N.J.), and genomic viewers utilizing BLAST homologies to sequenced and mapped genomic clones (NCBI, Bethesda, Md.).
184P1E2 maps to chromosome 1p36.13 using 184P1E2 sequence and the NCBI BLAST tool at the National Institutes of Health website.
Example 4
Expression Analysis of 184P1E2 in Normal Tissues and Patient Specimens
Expression analysis by RT-PCR demonstrated that 184P1E2 is strongly expressed in bladder cancer patient specimens (FIG. 14). First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P1E2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 184P1E2 in bladder cancer pool. Expression of 184P1E2 is also detected in kidney cancer pool, lung cancer pool and cancer metastasis pool but not in vital pool 1 and vital pool 2.
Extensive northern blot analysis of 184P1E2 in multiple human normal tissues is shown in FIG. 15. No expression was detected in all 16 normal tissues tested.
Expression of 184P1E2 in bladder cancer patient specimens and human normal tissues is shown in FIG. 16. RNA was extracted from a pool of three bladder cancers, as well as from normal prostate (NP), normal bladder (NB), normal kidney (NK), normal colon (NC), normal lung (NL) normal breast (NBr) and normal ovary (NO). Northern blot with 10 ug of total RNA/lane was probed with 184P1E2 sequence. The results show expression of an approximately 4.5 kb 184P1E2 transcript in the bladder cancer pool but not in the normal tissues tested. Analysis of individual patient specimens is shown in FIG. 17. RNA was extracted from normal bladder (NB), bladder cancer cell lines (CL; UM-UC-3, J82 and SCaBER), bladder cancer patient tumors (T) and normal tissue adjacent to bladder cancer (N). Northern blots with 10 ug of total RNA were probed with the 184P1E2 sequence. Size standards in kilobases are indicated on the side. Results show strong expression of 184P1E2 in the bladder tumor tissues but not in normal bladder, nor in the bladder cancer cell lines.
FIG. 18 shows that 184P1E2 was expressed in lung cancer patient tissues. RNA was extracted from normal lung (N), lung cancer cell lines (CALU-1, A427, NCI-H82, NCI-146) (all referred to as “CL”), lung cancer patient tumors (T) and their normal adjacent tissues (Nat). Northern blots with 10 ug of total RNA were probed with the 184P1E2 sequence. Results show strong expression of 184P1E2 in the lung cancer patient tissues, but not in normal lung. A lower molecular weight transcript of approximately 2.0 kb was also detected in the two lung cancer cell lines CALU-1 and NCI-H146.
The restricted expression of 184P1E2 in normal tissues and the expression detected in bladder cancer, lung cancer, kidney cancer, and cancer metastases suggest that 184P1E2 is a potential therapeutic target and a diagnostic marker for human cancers.
Example 5
Transcript Variants of 184P1E2
Transcript variants are variants of matured mRNA from the same gene by alternative transcription or alternative splicing. Alternative transcripts are transcripts from the same gene but start transcription at different points. Splice variants are mRNA variants spliced differently from the same transcript. In eukaryotes, when a multi-exon gene is transcribed from genomic DNA, the initial RNA is spliced to produce functional mRNA, which has only exons and is used for translation into an amino acid sequence. Accordingly, a given gene can have zero to many alternative transcripts and each transcript can have zero to many splice variants. Each transcript variant has a unique exon makeup, and can have different coding and/or non-coding (5′ or 3′ end) portions, from the original transcript. Transcript variants can code for similar or different proteins with the same or a similar function or may encode proteins with different functions, and may be expressed in the same tissue at the same time, or at different tissue, or at different times, proteins encoded by transcript variants can have similar or different cellular or extracellular localizations, i.e., be secreted.
Transcript variants are identified by a variety of art-accepted methods. For example, alternative transcripts and splice variants are identified in a full-length cloning experiment, or by use of full-length transcript and EST sequences. First, all human ESTs were grouped into clusters which show direct or indirect identity with each other. Second, ESTs in the same cluster were further grouped into sub-clusters and assembled into a consensus sequence. The original gene sequence is compared to the consensus sequence(s) or other full-length sequences. Each consensus sequence is a potential splice variant for that gene. Even when a variant is identified that is not a full-length clone, that portion of the variant is useful for antigen generation and for further cloning of the full-length splice variant, using techniques known in the art.
Moreover, computer programs are available in the art that identify transcript variants based on genomic sequences. Genomic-based transcript variant identification programs include FgenesH (A. Salamov and V. Solovyev, “Ab initio gene finding in Drosophila genomic DNA,” Genome Research. 2000 April; 10(4):516-22); Grail and GenScan on the INTERNET. For a general discussion of splice variant identification protocols see., e.g., Southan, C., A genomic perspective on human proteases, FEBS Lett. 2001 Jun. 8; 498(2-3):214-8; de Souza, S. J., et al., Identification of human chromosome 22 transcribed sequences with ORF expressed sequence tags, Proc. Natl. Acad Sci USA. 2000 Nov. 7; 97(23):12690-3.
To further confirm the parameters of a transcript variant, a variety of techniques are available in the art, such as full-length cloning, proteomic validation, PCR-based validation, and 5′ RACE validation, etc. (see e.g., Proteomic Validation Brennan, S. O., et al., Albumin banks peninsula: a new termination variant characterized by electrospray mass spectrometry, Biochem Biophys Acta. 1999 Aug. 17; 1433(1-2):321-6; Ferranti P, et al., Differential splicing of pre-messenger RNA produces multiple forms of mature caprine alpha(s1)-casein, Eur J. Biochem. 1997 Oct. 1; 249(1):1-7. For PCR-based Validation: Wellmann S, et al., Specific reverse transcription-PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler technology, Clin Chem. 2001 April; 47(4):654-60; Jia, H. P., et al., Discovery of new human beta-defensins using a genomics-based approach, Gene. 2001 Jan. 24; 263(1-2):211-8. For PCR-based and 5′ RACE Validation: Brigle, K. E., et al., Organization of the murine reduced folate carrier gene and identification of variant splice forms, Biochem Biophys Acta. 1997 Aug. 7; 1353(2): 191-8).
It is known in the art that genomic regions are modulated in cancers. When the genomic region, to which a gene maps, is modulated in a particular cancer, the alternative transcripts or splice variants of the gene are modulated as well. Disclosed herein is that 184P1E2 has a particular expression profile related to cancer. Alternative transcripts and splice variants of 184P1E2 may also be involved in cancers in the same or different tissues, thus serving as tumor-associated markers/antigens.
The exon composition of the original transcript, designated as 184P1E2 v.1, is shown in FIG. 12.
Example 6
Single Nucleotide Polymorphisms of 184P1E2
A Single Nucleotide Polymorphism (SNP) is a single base pair variation in nucleotide sequences. At a specific point of the genome, there are four possible nucleotide base pairs: A/T, C/G, G/C and T/A. Genotype refers to the base pair make-up of one or more spots in the genome of an individual, while haplotype refers to base pair make-up of more than one varied spots on the same DNA molecule (chromosome in higher organism). SNPs that occur on a cDNA are called cSNPs. These cSNPs may change amino acids of the protein encoded by the gene and thus change the functions of the protein. Some SNPs cause inherited diseases and some others contribute to quantitative variations in phenotype and reactions to environmental factors including diet and drugs among individuals. Therefore, SNPs and/or combinations of alleles (called haplotypes) have many applications including diagnosis of inherited diseases, determination of drug reactions and dosage, identification of genes responsible for disearses and discovery of genetic relationship between individuals (P. Nowotny, J. M. Kwon and A. M. Goate, “SNP analysis to dissect human traits,” Curr. Opin. Neurobiol. 2001 October; 11(5):637-641; M. Pirmohamed and B. K. Park, “Genetic susceptibility to adverse drug reactions,” Trends Pharmacol. Sci. 2001 June; 22(6):298-305; J. H. Riley, C. J. Allan, E. Lai and A. Roses, “The use of single nucleotide polymorphisms in the isolation of common disease genes,” Pharmacogenomics. 2000 February; 1(1):39-47; R. Judson, J. C. Stephens and A. Windemuth, “The predictive power of haplotypes in clinical response,” Pharmacogenomics. 2000 feb; 1(1):15-26).
SNPs are identified by a variety of art-accepted methods (P. Bean, “The promising voyage of SNP target discovery,” Am. Clin. Lab. 2001 October-November; 20(9):18-20; K. M. Weiss, “In search of human variation,” Genome Res. 1998 July; 8(7):691-697; M. M. She, “Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies,” Clin. Chem. 2001 February; 47(2): 164-172). For example, SNPs are identified by sequencing DNA fragments that show polymorphism by gel-based methods such as restriction fragment length polymorphism (RFLP) and denaturing gradient gel electrophoresis (DGGE). They can also be discovered by direct sequencing of DNA samples pooled from different individuals or by comparing sequences from different DNA samples. With the rapid accumulation of sequence data in public and private databases, one can discover SNPs by comparing sequences using computer programs (Z. Gu, L. Hillier and P. Y. Kwok, “Single nucleotide polymorphism hunting in cyberspace,” Hum. Mutat. 1998; 12(4):221-225). SNPs can be verified and genotype or haplotype of an individual can be determined by a variety of methods including direct sequencing and high throughput microarrays (P. Y. Kwok, “Methods for genotyping single nucleotide polymorphisms,” Annu. Rev. Genomics Hum. Genet. 2001; 2:235-258; M. Kokoris, K. Dix, K. Moynihan, J. Mathis, B. Erwin, P. Grass, B. Hines and A. Duesterhoeft, “High-throughput SNP genotyping with the Masscode system,” Mol. Diagn. 2000 December; 5(4):329-340).
Using the methods described above, nine SNPs were identified in the transcript, 184P1E2 v.1, at positions 951 (C/G), 1480 (C/T), 1910 (T/G), 2468 (C/T), 2623 (T/G), 2742 (G/T), 2924 (A/C), 3060 (C/A) and 356 (G/A). The transcripts with alternative alleles were designated as variants 184P1E2 v.2, v.3, v.4, v.5, v.6, v.7, v.8, v.9, and v.10, respectively. FIG. 10 shows the schematic alignment of the nucleotide variants. FIG. 11 shows the schematic alignment of protein variants, corresponding to nucleotide variants. Nucleotide variants that code for the same amino acid sequence as variant 1 are not shown in FIG. 11. These alleles of the SNPs, though shown separately here, can occur in different combinations (haplotypes) and in any other transcript variants that contains the sequence context of the SNPs.
Example 7
Production of Recombinant 184P1E2 in Prokaryotic Systems
To express recombinant 184P1E2 and 184P1E2 variants in prokaryotic cells, the full or partial length 184P1E2 and 184P1E2 variant cDNA sequences are cloned into any one of a variety of expression vectors known in the art. One or more of the following regions of 184P1E2 amino acids 1-664; or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids from 184P1E2, variants, or analogs thereof.
A. In Vitro Transcription and Translation Constructs:
pCRII: To generate 184P1E2 sense and anti-sense RNA probes for RNA in situ investigations, pCRII constructs (Invitrogen, Carlsbad Calif.) are generated encoding either all or fragments of the 184P1E2 cDNA. The pCRII vector has Sp6 and T7 promoters flanking the insert to drive the transcription of 184P1E2 RNA for use as probes in RNA in situ hybridization experiments. These probes are used to analyze the cell and tissue expression of 184P1E2 at the RNA level. Transcribed 184P1E2 RNA representing the cDNA amino acid coding region of the 184P1E2 gene is used in in vitro translation systems such as the TnTTM Coupled Reticulolysate System (Promega, Corp., Madison, Wis.) to synthesize 184P1E2 protein.
B. Bacterial Constructs:
pGEX Constructs: To generate recombinant 184P1E2 proteins in bacteria that are fused to the Glutathione S-transferase (GST) protein, all or parts of the T-fusion vector of the pGEX family (Amersham Pharmacia Biotech, Piscataway, N.J.). These constructs allow controlled expression of recombinant 184P1E2 protein sequences with GST fused at the amino-terminus and a six histidine epitope (6×His) at the carboxyl-terminus. The GST and 6×His tags permit purification of the recombinant fusion protein from induced bacteria with the appropriate affinity matrix and allow recognition of the fusion protein with anti-GST and anti-His antibodies. The 6×His tag is generated by adding 6 histidine codons to the cloning primer at the 3′ end, e.g., of the open reading frame (ORF). A proteolytic cleavage site, such as the PreScissionTM recognition site in pGEX-6P-1, may be employed such that it permits cleavage of the GST tag from 184P1E2-related protein. The ampicillin resistance gene and pBR322 origin permits selection and maintenance of the pGEX plasmids in E. coli. 
pMAL Constructs: To generate, in bacteria, recombinant 184P1E2 proteins that are fused to maltose-binding protein (MBP), all or parts of the 184P1E2 cDNA protein coding sequence are fused to the MBP gene by cloning into the pMAL-c2X and pMAL-p2X vectors (New England Biolabs, Beverly, Mass.). These constructs allow controlled expression of recombinant 184P1E2 protein sequences with MBP fused at the amino-terminus and a 6×His epitope tag at the carboxyl-terminus. The MBP and 6×His tags permit purification of the recombinant protein from induced bacteria with the appropriate affinity matrix and allow recognition of the fusion protein with anti-MBP and anti-His antibodies. The 6×His epitope tag is generated by adding 6 histidine codons to the 3′ cloning primer. A Factor Xa recognition site permits cleavage of the pMAL tag from 184P1E2. The pMAL-c2X and pMAL-p2X vectors are optimized to express the recombinant protein in the cytoplasm or periplasm respectively. Periplasm expression enhances folding of proteins with disulfide bonds.
pET Constructs: To express 184P1E2 in bacterial cells, all or parts of the 184P1E2 cDNA protein coding sequence are cloned into the pET family of vectors (Novagen, Madison, Wis.). These vectors allow tightly controlled expression of recombinant 184P1E2 protein in bacteria with and without fusion to proteins that enhance solubility, such as NusA and thioredoxin (Trx), and epitope tags, such as 6×His and S-TagTM that aid purification and detection of the recombinant protein. For example, constructs are made utilizing pET NusA fusion system 43.1 such that regions of the 184P1E2 protein are expressed as amino-terminal fusions to NusA.
C. Yeast Constructs:
pESC Constructs: To express 184P1E2 in the yeast species Saccharomyces cerevisiae for generation of recombinant protein and functional studies, all or parts of the 184P1E2 cDNA protein coding sequence are cloned into the pESC family of vectors each of which contain 1 of 4 selectable markers, HIS3, TRP1, LEU2, and URA3 (Stratagene, La Jolla, Calif.). These vectors allow controlled expression from the same plasmid of up to 2 different genes or cloned sequences containing either FlagTM or Myc epitope tags in the same yeast cell. This system is useful to confirm protein-protein interactions of 184P1E2. In addition, expression in yeast yields similar post-translational modifications, such as glycosylations and phosphorylations, that are found when expressed in eukaryotic cells.
pESP Constructs: To express 184P1E2 in the yeast species Saccharomyces pombe, all or parts of the 184P1E2 cDNA protein coding sequence are cloned into the pESP family of vectors. These vectors allow controlled high level of expression of a 184P1E2 protein sequence that is fused at either the amino terminus or at the carboxyl terminus to GST which aids purification of the recombinant protein. A FlagTM epitope tag allows detection of the recombinant protein with anti-FlagTM antibody.
Example 8
Production of Recombinant 184P1E2 in Eukaryotic Systems
A. Mammalian Constructs:
To express recombinant 184P1E2 in eukaryotic cells, the full or partial length 184P1E2 cDNA sequences can be cloned into any one of a variety of expression vectors . known in the art. One or more of the following regions of 184P1E2 are expressed in these constructs, amino acids 1 to 664, or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or more contiguous amino acids from 184P1E2, variants, or analogs thereof. In certain embodiments a region of a specific variant of 184P1E2 is expressed that encodes an amino acid at a specific position which differs from the amino acid of any other variant found at that position. In other embodiments, a region of a variant of 184P1E2 is expressed that lies partly or entirely within a sequence that is unique to that variant.
The constructs can be transfected into any one of a wide variety of mammalian cells such as 293T cells. Transfected 293T cell lysates can be probed with the anti-184P1E2 polyclonal serum, described herein.
pcDNA4/HisMax Constructs: To express 184P1E2 in mammalian cells, a 184P1E2 ORF, or portions thereof, of 184P1E2 are cloned into pcDNA4/HisMax Version A (Invitrogen, Carlsbad, Calif.). Protein expression is driven from the cytomegalovirus (CMV) promoter and the SP16 translational enhancer. The recombinant protein has XpressTM and six histidine (6×His) epitopes fused to the amino-terminus. The pcDNA4/HisMax vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Zeocin resistance gene allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli. 
pcDNA3.1/MycHis Constructs: To express 184P1E2 in mammalian cells, a 184P1E2 ORF, or portions thereof, of 184P1E2 with a consensus Kozak translation initiation site are cloned into pcDNA3.1/MycHis Version A (Invitrogen, Carlsbad, Calif.). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the myc epitope and 6×His epitope fused to the carboxyl-terminus. The pcDNA3.1/MycHis vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability, along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene can be used, as it allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli. 
pcDNA3.1/CT-GFP-TOPO Construct: To express 184P1E2 in mammalian cells and to allow detection of the recombinant proteins using fluorescence, a 184P1E2 ORF, or portions thereof, with a consensus Kozak translation initiation site are cloned into pcDNA3.1/CT-GFP-TOPO (Invitrogen, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the Green Fluorescent Protein (GFP) fused to the carboxyl-terminus facilitating non-invasive, in vivo detection and cell biology studies. The pcDNA3.1CT-GFP-TOPO vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. coli. Additional constructs with an amino-terminal GFP fusion are made in pcDNA3.1/NT-GFP-TOPO spanning the entire length of a 184P1E2 protein.
PAPtag: A 184P1E2 ORF, or portions thereof, is cloned into pAPtag-5 (GenHunter Corp. Nashville, Tenn.). This construct generates an alkaline phosphatase fusion at the carboxyl-terminus of a 184P1E2 protein while fusing the IgGκ signal sequence to the amino-terminus. Constructs are also generated in which alkaline phosphatase with an amino-terminal IgGK signal sequence is fused to the amino-terminus of a 184P1E2 protein. The resulting recombinant 184P1E2 proteins are optimized for secretion into the media of transfected mammalian cells and can be used to identify proteins such as ligands or receptors that interact with 184P1E2 proteins. Protein expression is driven from the CMV promoter and the recombinant proteins also contain myc and 6×His epitopes fused at the carboxyl-terminus that facilitates detection and purification. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the recombinant protein and the ampicillin resistance gene permits selection of the plasmid in E. coli. 
ptag5: A 184P1E2 ORF, or portions thereof, is cloned into pTag-5. This vector is similar to pAPtag but without the alkaline phosphatase fusion. This construct generates 184P1E2 protein with an amino-terminal IgGκ signal sequence and myc and 6×His epitope tags at the carboxyl-terminus that facilitate detection and affinity purification. The resulting recombinant 184P1E2 protein is optimized for secretion into the media of transfected mammalian cells, and is used as immunogen or ligand to identify proteins such as ligands or receptors that interact with the 184P1E2 proteins. Protein expression is driven from the CMV promoter. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the protein, and the ampicillin resistance gene permits selection of the plasmid in E. coli. 
PsecFe: A 184P1E2 ORF, or portions thereof, is also cloned into psecFc. The psecFc vector was assembled by cloning the human immunoglobulin G1 (IgG) Fc (hinge, CH2, CH3 regions) into pSecTag2 (Invitrogen, California). This construct generates an IgG1 Fc fusion at the carboxyl-terminus of the 184P1E2 proteins, while fusing the IgGK signal sequence to N-terminus. 184P1E2 fusions utilizing the murine IgG1 Fc region are also used. The resulting recombinant 184P1E2 proteins are optimized for secretion into the media of transfected mammalian cells, and can be used as immunogens or to identify proteins such as ligands or receptors that interact with 184P1E2 protein. Protein expression is driven from the CMV promoter. The hygromycin resistance gene present in the vector allows for selection of mammalian cells that express the recombinant protein, and the ampicillin resistance gene permits selection of the plasmid in E. coli. 
pSRα Constructs: To generate mammalian cell lines that express 184P1E2 constitutively, 184P1E2 ORF, or portions thereof, of 184P1E2 are cloned into pSRα constructs. Amphotropic and ecotropic retroviruses are generated by transfection of pSRα constructs into the 293T-10A1 packaging line or co-transfection of pSRα and a helper plasmid (containing deleted packaging sequences) into the 293 cells, respectively. The retrovirus is used to infect a variety of mammalian cell lines, resulting in the integration of the cloned gene, 184P1E2, into the host cell-lines. Protein expression is driven from a long terminal repeat (LTR). The Neomycin resistance gene present in the vector allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColE1 origin permit selection and maintenance of the plasmid in E. coli. The retroviral vectors can thereafter be used for infection and generation of various cell lines using, for example, PC3, NIH 3T3, TsuPr1, 293 or rat-1 cells.
Additional pSRα constructs are made that fuse an epitope tag such as the FLAGTM tag to the carboxyl-terminus of 184P1E2 sequences to allow detection using anti-Flag antibodies. For example, the FLAGTM sequence 5′ gat tac aag gat gac gac gat aag 3′ (SEQ ID NO: 45) is added to cloning primer at the 3′ end of the ORF. Additional pSRα constructs are made to produce both amino-terminal and carboxyl-terminal GFP and myc/6×His fusion proteins of the full-length 184P1E2 proteins.
Additional Viral Vectors: Additional constructs are made for viral-mediated delivery and expression of 184P1E2. High virus titer leading to high level expression of 184P1E2 is achieved in viral delivery systems such as adenoviral vectors and herpes amplicon vectors. A 184P1E2 coding sequences or fragments thereof are amplified by PCR and subcloned into the AdEasy shuttle vector (Stratagene). Recombination and virus packaging are performed according to the manufacturer's instructions to generate adenoviral vectors. Alternatively, 184P1E2 coding sequences or fragments thereof are cloned into the HSV-1 vector (Imgenex) to generate herpes viral vectors. The viral vectors are thereafter used for infection of various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.
Regulated Expression Systems: To control expression of 184P1E2 in mammalian cells, coding sequences of 184P1E2, or portions thereof, are cloned into regulated mammalian expression systems such as the T-Rex System (Invitrogen), the GeneSwitch System (Invitrogen) and the tightly-regulated Ecdysone System (Sratagene). These systems allow the study of the temporal and concentration dependent effects of recombinant 184P1E2. These vectors are thereafter used to control expression of 184P1E2 in various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.
B. Baculovirus Expression Systems
To generate recombinant 184P1E2 proteins in a baculovirus expression system, 184P1E2 ORF, or portions thereof, are cloned into the baculovirus transfer vector pBlueBac 4.5 (Invitrogen), which provides a His-tag at the N-terminus. Specifically, pBlueBac-184P1E2 is co-transfected with helper plasmid pBac-N-Blue (Invitrogen) into SF9 (Spodoptera frugiperda) insect cells to generate recombinant baculovirus (see Invitrogen instruction manual for details). Baculovirus is then collected from cell supernatant and purified by plaque assay.
Recombinant 184P1E2 protein is then generated by infection of HighFive insect cells (Invitrogen) with purified baculovirus. Recombinant 184P1E2 protein can be detected using anti-184P1E2 or anti-His-tag antibody. 184P1E2 protein can be purified and used in various cell-based assays or as immunogen to generate polyclonal and monoclonal antibodies specific for 184P1E2.
Example 9
Antigenicity Profiles and Secondary Structure
FIG. 5, FIG. 6, FIG. 7, FIG. 8, and FIG. 9 depict graphically five amino acid profiles of the 184P1E2 variants 1 through 4 respectively, each assessment available by accessing the ProtScale website on the ExPasy molecular biology server.
These profiles: FIG. 5, Hydrophilicity, (Hopp T. P., Woods K. R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828); FIG. 6, Hydropathicity, (Kyte J., Doolittle R. F., 1982. J. Mol. Biol. 157:105-132); FIG. 7, Percentage Accessible Residues (Janin J., 1979 Nature 277:491-492); FIG. 8, Average Flexibility, (Bhaskaran R., and Ponnuswamy P. K., 1988. Int. J. Pept. Protein Res. 32:242-255); FIG. 9, Beta-turn (Deleage, G., Roux B. 1987 Protein Engineering 1:289-294); and optionally others available in the art, such as on the ProtScale website, were used to identify antigenic regions of the 184P1E2 protein. Each of the above amino acid profiles of 184P1E2 were generated using the following ProtScale parameters for analysis: 1) A window size of 9; 2) 100% weight of the window edges compared to the window center; and, 3) amino acid profile values normalized to lie between 0 and 1.
Hydrophilicity (FIG. 5), Hydropathicity (FIG. 6) and Percentage Accessible Residues (FIG. 7) profiles were used to determine stretches of hydrophilic amino acids (i.e., values greater than 0.5 on the Hydrophilicity and Percentage Accessible Residues profile, and values less than 0.5 on the Hydropathicity profile). Such regions are likely to be exposed to the aqueous environment, be present on the surface of the protein, and thus available for immune recognition, such as by antibodies.
Average Flexibility (FIG. 8) and Beta-turn (FIG. 9) profiles determine stretches of amino acids (i.e., values greater than 0.5 on the Beta-turn profile and the Average Flexibility profile) that are not constrained in secondary structures such as beta sheets and alpha helices. Such regions are also more likely to be exposed on the protein and thus accessible to immune recognition, such as by antibodies.
Antigenic sequences of the 184P1E2 protein and of the variant proteins indicated, e.g., by the profiles set forth in FIG. 5, FIG. 6, FIG. 7, FIG. 8, and/or FIG. 9 are used to prepare immunogens, either peptides or nucleic acids that encode them, to generate therapeutic and diagnostic anti-184P1E2 antibodies. The immunogen can be any 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50 or more than 50 contiguous amino acids, or the corresponding nucleic acids that encode them, from the 184P1E2 protein variants listed in FIGS. 2 and 3. In particular, peptide immunogens of the invention can comprise, a peptide region of at least 5 amino acids of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of FIG. 5; a peptide region of at least 5 amino acids of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of FIG. 6 ; a peptide region of at least 5 amino acids of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of FIG. 7 ; a peptide region of at least 5 amino acids of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profile on FIG. 8; and, a peptide region of at least 5 amino acids of FIGS. 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of FIG. 9 . Peptide immunogens of the invention can also comprise nucleic acids that encode any of the forgoing.
All immunogens of the invention, peptide or nucleic acid, can be embodied in human unit dose form, or comprised by a composition that includes a pharmaceutical excipient compatible with human physiology.
The secondary structure of 184P1E2 variant 1, namely the predicted presence and location of alpha helices, extended strands, and random coils, is predicted from the primary amino acid sequence using the HNN—Hierarchical Neural Network method (Guermeur, 1997, accessed from the ExPasy molecular biology server. The analysis indicates that 184P1E2 variant 1 is composed of 25.30% alpha helix, 22.59% extended strand, and 52.11% random coil (FIG. 13).
Analysis for the potential presence of transmembrane domains in 184P1E2 variant 1 was carried out using a variety of transmembrane prediction algorithms accessed from the ExPasy molecular biology server. The programs do not predict the presence of transmembrane domains in 184P1E2, suggesting that it is a soluble protein.
Example 10
Generation of 184P1E2 Polyclonal Antibodies
Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. In addition to immunizing with the full length 184P1E2 protein, computer algorithms are employed in design of immunogens that, based on amino acid sequence analysis contain characteristics of being antigenic and available for recognition by the immune system of the immunized host (see the Example entitled “Antigenicity Profiles”). Such regions would be predicted to be hydrophilic, flexible, in beta-turn conformations, and be exposed on the surface of the protein (see, e.g., FIG. 5, FIG. 6, FIG. 7, FIG. 8, or FIG. 9 for amino acid profiles that indicate such regions of 184P1E2 and variants).
For example, 184P1E2 recombinant bacterial fusion proteins or peptides containing hydrophilic, flexible, beta-turn regions of 184P1E2 variant proteins are used as antigens to generate polyclonal antibodies in New Zealand White rabbits. For example, such regions include, but are not limited to, amino acids 53-73, amino acids, amino acids 117-136, amino acids 217-251, and 366-446 of 184P1E2 variant 1. It is useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. In one embodiment, a peptide encoding amino acids 53-73 of 184P1E2 variant 1 is conjugated to KLH and used to immunize the rabbit. Alternatively the immunizing agent may include all or portions of the 184P1E2 variant proteins, analogs or fusion proteins thereof. For example, the 184P1E2 variant 1 amino acid sequence can be fused using recombinant DNA techniques to any one of a variety of fusion protein partners that are well known in the art, such as glutathione-S-transferase (GST) and HIS tagged fusion proteins. Such fusion proteins are purified from induced bacteria using the appropriate affinity matrix.
In one embodiment, a GST-fusion protein encoding amino acids 1-251, encompassing several predicted antigenic regions, is produced and purified and used as immunogen. Other recombinant bacterial fusion proteins that may be employed include maltose binding protein, LacZ, thioredoxin, NusA, or an immunoglobulin constant region (see the section entitled “Production of 184P1E2 in Prokaryotic Systems” and Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubul et al. eds., 1995; Linsley, P. S., Brady, W., Urnes, M., Grosmaire, L., Damle, N., and Ledbetter, L. (1991) J. Exp. Med. 174, 561-566).
In addition to bacterial derived fusion proteins, mammalian expressed protein antigens are also used. These antigens are expressed from mammalian expression vectors such as the Tag5 and Fc-fusion vectors (see the Example entitled “Production of Recombinant 184P1E2 in Eukaryotic Systems”), and retain post-translational modifications such as glycosylations found in native protein. In one embodiment, the full length sequence of variant 1, amino acids 1-664, is cloned into the Tag5 mammalian secretion vector. The recombinant protein is purified by metal chelate chromatography from tissue culture supernatants of 293T cells stably expressing the recombinant vector. The purified Tag5 184P1E2 protein is then used as immunogen.
During the immunization protocol, it is useful to mix or emulsify the antigen in adjuvants that enhance the immune response of the host animal. Examples of adjuvants include, but are not limited to, complete Freund's adjuvant (CFA) and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).
In a typical protocol, rabbits are initially immunized subcutaneously with up to 200 μg, typically 100-200 μg, of fusion protein or peptide conjugated to KLH mixed in complete Freund's adjuvant (CFA). Rabbits are then injected subcutaneously every two weeks with up to 200 μg, typically 100-200 μg, of the immunogen in incomplete Freund's adjuvant (IFA). Test bleeds are taken approximately 7-10 days following each immunization and used to monitor the titer of the antiserum by ELISA.
To test reactivity and specificity of immune serum, such as the rabbit serum derived from immunization with a KLH-conjugated peptide encoding amino acids 53-73 of variant 1, the full-length 184P1E2 variant 1 cDNA is cloned into pcDNA 3.1 myc-his expression vector (Invitrogen, see the Example entitled “Production of Recombinant 184P1E2 in Eukaryotic Systems”). After transfection of the constructs into 293T cells, cell lysates are probed with the anti-184P1E2 serum and with anti-His antibody (Santa Cruz Biotechnologies, Santa Cruz, Calif.) to determine specific reactivity to denatured 184P1E2 protein using the Western blot technique. The immune serum is then tested by the Western blot technique against 293T-184P1E2 cells. In addition, the immune serum is tested by fluorescence microscopy, flow cytometry and immunoprecipitation against 293T and other recombinant 184P1E2-expressing cells to determine specific recognition of native protein. Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometric techniques using cells that endogenously express 184P1E2 are also carried out to test reactivity and specificity.
Anti-serum from rabbits immunized with 184P1E2 variant fusion proteins, such as GST and MBP fusion proteins, are purified by depletion of antibodies reactive to the fusion partner sequence by passage over an affinity column containing the fusion partner either alone or in the context of an irrelevant fusion protein. For example, antiserum derived from a GST-184P1E2 fusion protein encoding amino acids 1-251 is first purified by passage over a column of GST protein covalently coupled to AffiGel matrix (BioRad, Hercules, Calif.). The antiserum is then affinity purified by passage over a column composed of a MBP-fusion protein also encoding amino acids 1-251 covalently coupled to Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG fraction. Sera from other His-tagged antigens and peptide immunized rabbits as well as fusion partner depleted sera are affinity purified by passage over a column matrix composed of the original protein immunogen or free peptide.
Example 11
Generation of 184P1E2 Monoclonal Antibodies (mAbs)
In one embodiment, therapeutic mAbs to 184P1E2 variants comprise those that react with epitopes specific for each variant protein or specific to sequences in common between the variants that would disrupt or modulate the biological function of the 184P1E2 variants, for example those that would disrupt the interaction with ligands and substrates or disrupt its catalytic activity. Immunogens for generation of such mAbs include those designed to encode or contain the entire 184P1E2 protein variant sequence, regions of the 184P1E2 protein variants predicted to be antigenic from computer analysis of the amino acid sequence (see, e.g., FIG. 5, FIG. 6, FIG. 7, FIG. 8, or FIG. 9, and the Example entitled “Antigenicity Profiles”). Immunogens include peptides, recombinant bacterial proteins, and mammalian expressed Tag 5 proteins and human and murine IgG FC fusion proteins. In addition, cells engineered to express high levels of a respective 184P1E2 variant, such as 293T-184P1E2 variant 1 or 300.19-184P1E2 variant Imurine Pre-B cells, are used to immunize mice.
To generate mAbs to a 184P1E2 variant, mice are first immunized intraperitoneally (IP) with, typically, 10-50 μg of protein immunogen or 107 184P1E2-expressing cells mixed in complete Freund's adjuvant. Mice are then subsequently immunized IP every 2-4 weeks with, typically, 10-50 μg of protein immunogen or 107 cells mixed in incomplete Freund's adjuvant. Alternatively, MPL-TDM adjuvant is used in immunizations. In addition to the above protein and cell-based immunization strategies, a DNA-based immunization protocol is employed in which a mammalian expression vector encoding a 184P1E2 variant sequence is used to immunize mice by direct injection of the plasmid DNA. For example, the full length variant 1 sequence, encoding amino acids 1-664, is cloned into the Tag5 mammalian secretion vector and the recombinant vector is used as immunogen. In another example the same amino acids are cloned into an Fc-fusion secretion vector in which the 184P1E2 variant 1 sequence is fused at the amino-terminus to an IgK leader sequence and at the carboxyl-terminus to the coding sequence of the human or murine IgG Fc region. This recombinant vector is then used as immunogen. The plasmid immunization protocols are used in combination with purified proteins expressed from the same vector and with cells expressing the respective 184P1E2 variant.
During the immunization protocol, test bleeds are taken 7-10 days following an injection to monitor titer and specificity of the immune response. Once appropriate reactivity and specificity is obtained as determined by ELISA, Western blotting, immunoprecipitation, fluorescence microscopy, and flow cytometric analyses, fusion and hybridoma generation is then carried out with established procedures well known in the art (see, e.g., Harlow and Lane, 1988).
In one embodiment for generating 184P1E2 monoclonal antibodies, a Tag5-184P1E2 variant 1 antigen encoding amino acids 1-664, is expressed and purified from stably transfected 293T cells. Balb C mice are initially immunized intraperitoneally with 25 μg of the Tag5-184P1E2 variant 1 protein mixed in complete Freund's adjuvant. Mice are subsequently immunized every two weeks with 25 μg of the antigen mixed in incomplete Freund's adjuvant for a total of three immunizations. ELISA using the Tag5 antigen determines the titer of serum from immunized mice. Reactivity and specificity of serum to full length 184P1E2 variant protein is monitored by Western blotting, immunoprecipitation and flow cytometry using 293T cells transfected with an expression vector encoding the 184P1E2 variant 1 cDNA (see e.g., the Example entitled “Production of Recombinant 184P1E2 in Eukaryotic Systems”). Other recombinant 184P1E2 variant 1-expressing cells or cells endogenously expressing 184P1E2 variant 1 are also used. Mice showing the strongest reactivity are rested and given a final injection of Tag5 antigen in PBS and then sacrificed four days later. The spleens of the sacrificed mice are harvested and fused to SPO/2 myeloma cells using standard procedures (Harlow and Lane, 1988). Supernatants from HAT selected growth wells are screened by ELISA, Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometry to identify 184P1E2 specific antibody-producing clones.
The binding affinity of a 184P1E2 monoclonal antibody is determined using standard technologies. Affinity measurements quantify the strength of antibody to epitope binding and are used to help define which 184P1E2 monoclonal antibodies preferred for diagnostic or therapeutic use, as appreciated by one of skill in the art. The BIAcore system (Uppsala, Sweden) is a preferred method for determining binding affinity. The BIAcore system uses surface plasmon resonance (SPR, Welford K. 1991, Opt. Quant. Elect. 23:1; Morton and Myszka, 1998, Methods in Enzymology 295: 268) to monitor biomolecular interactions in real time. BIAcore analysis conveniently generates association rate constants, dissociation rate constants, equilibrium dissociation constants, and affinity constants.
Example 12
HLA Class I and Class II Binding Assays
HLA class I and class II binding assays using purified HLA molecules are performed in accordance with disclosed protocols (e.g., PCT publications WO 94/20127 and WO 94/03205; Sidney et al., Current Protocols in Immunology 18.3.1 (1998); Sidney, et al., J. Immunol. 154:247 (1995); Sette, et al., Mol. Immunol. 31:813 (1994)). Briefly, purified MHC molecules (5 to 500 nM) are incubated with various unlabeled peptide inhibitors and 1-10 nM 125I-radiolabeled probe peptides as described. Following incubation, MHC-peptide complexes are separated from free peptide by gel filtration and the fraction of peptide bound is determined. Typically, in preliminary experiments, each MHC preparation is titered in the presence of fixed amounts of radiolabeled peptides to determine the concentration of HLA molecules necessary to bind 10-20% of the total radioactivity. All subsequent inhibition and direct binding assays are performed using these HLA concentrations.
Since under these conditions [label]<[HLA] and IC50≧[HLA], the measured IC50 values are reasonable approximations of the true KD values. Peptide inhibitors are typically tested at concentrations ranging from 120 μg/ml to 1.2 ng/ml, and are tested in two to four completely independent experiments. To allow comparison of the data obtained in different experiments, a relative binding figure is calculated for each peptide by dividing the IC50 of a positive control for inhibition by the IC50 for each tested peptide (typically unlabeled versions of the radiolabeled probe peptide). For database purposes, and inter-experiment comparisons, relative binding values are compiled. These values can subsequently be converted back into IC50 nM values by dividing the IC50 nM of the positive controls for inhibition by the relative binding of the peptide of interest. This method of data compilation is accurate and consistent for comparing peptides that have been tested on different days, or with different lots of purified MHC.
Binding assays as outlined above may be used to analyze HLA supermotif and/or HLA motif-bearing peptides (see Table IV).
Example 13
Identification of HLA Supermotif- and Motif-Bearing CTL Candidate Epitopes
HLA vaccine compositions of the invention can include multiple epitopes. The multiple epitopes can comprise multiple HLA supermotifs or motifs to achieve broad population coverage. This example illustrates the identification and confirmation of supermotif- and motif-bearing epitopes for the inclusion in such a vaccine composition. Calculation of population coverage is performed using the strategy described below.
Computer Searches and Algorithms for Identification of Supermotif and/or Motif-Bearing Epitopes
The searches performed to identify the motif-bearing peptide sequences in the Example entitled “Antigenicity Profiles” and Tables V-XVIII and XXII-LI employ the protein sequence data from the gene product of 184P1E2 set forth in FIGS. 2 and 3, the specific peptides used to generate the tables are listed in table LII.
Computer searches for epitopes bearing HLA Class I or Class II supermotifs or motifs are performed as follows. All translated 184P1E2 protein sequences are analyzed using a text string search software program to identify potential peptide sequences containing appropriate HLA binding motifs; such programs are readily produced in accordance with information in the art in view of known motif/supermotif disclosures. Furthermore, such calculations can be made mentally.
Identified A2-, A3-, and DR-supermotif sequences are scored using polynomial algorithms to predict their capacity to bind to specific HLA-Class I or Class II molecules. These polynomial algorithms account for the impact of different amino acids at different positions, and are essentially based on the premise that the overall affinity (or ΔG) of peptide-HLA molecule interactions can be approximated as a linear polynomial function of the type:

“ΔG”=a1i×a2i×a3i . . . ×ani 

where aji is a coefficient which represents the effect of the presence of a given amino acid (j) at a given position (i) along the sequence of a peptide of n amino acids. The crucial assumption of this method is that the effects at each position are essentially independent of each other (i.e., independent binding of individual side-chains). When residue j occurs at position i in the peptide, it is assumed to contribute a constant amount ji to the free energy of binding of the peptide irrespective of the sequence of the rest of the peptide.
The method of derivation of specific algorithm coefficients has been described in Gulukota et al., J. Mol. Biol. 267:1258-126, 1997; (see also Sidney et al., Human Immunol. 45:79-93, 1996; and Southwood et al., J. Immunol. 160:3363-3373, 1998). Briefly, for all i positions, anchor and non-anchor alike, the geometric mean of the average relative binding (ARB) of all peptides carrying j is calculated relative to the remainder of the group, and used as the estimate of ji. For Class II peptides, if multiple alignments are possible, only the highest scoring alignment is utilized, following an iterative procedure. To calculate an algorithm score of a given peptide in a test set, the ARB values corresponding to the sequence of the peptide are multiplied. If this product exceeds a chosen threshold, the peptide is predicted to bind. Appropriate thresholds are chosen as a function of the degree of stringency of prediction desired.
Selection of HLA-A2 Supertype Cross-Reactive Peptides
Protein sequences from 184P1E2 are scanned utilizing motif identification software, to identify 8-, 9-10- and 11-mer sequences containing the HLA-A2-supermotif main anchor specificity. Typically, these sequences are then scored using the protocol described above and the peptides corresponding to the positive-scoring sequences are synthesized and tested for their capacity to bind purified HLA-A*0201 molecules in vitro (HLA-A*0201 is considered a prototype A2 supertype molecule).
These peptides are then tested for the capacity to bind to additional A2-supertype molecules (A*0202, A*0203, A*0206, and A*6802). Peptides that bind to at least three of the five A2-supertype alleles tested are typically deemed A2-supertype cross-reactive binders. Preferred peptides bind at an affinity equal to or less than 500 nM to three or more HLA-A2 supertype molecules.
Selection of HLA-A3 Supermotif-Bearing Epitopes
The 184P1E2 protein sequence(s) scanned above is also examined for the presence of peptides with the HLA-A3-supermotif primary anchors. Peptides corresponding to the HLA A3 supermotif-bearing sequences are then synthesized and tested for binding to HLA-A*0301 and HLA-A*1101 molecules, the molecules encoded by the two most prevalent A3-supertype alleles. The peptides that bind at least one of the two alleles with binding affinities of ≦500 nM, often ≦200 nM, are then tested for binding cross-reactivity to the other common A3-supertype alleles (e.g., A*3101, A*3301, and A*6801) to identify those that can bind at least three of the five HLA-A3-supertype molecules tested.
Selection of HLA-B7 Supermotif Bearing Epitopes
The 184P1E2 protein(s) scanned above is also analyzed for the presence of 8-, 9-10-, or 1 1-mer peptides with the HLA-B7-supermotif. Corresponding peptides are synthesized and tested for binding to HLA-B*0702, the molecule encoded by the most common B7-supertype allele (i.e., the prototype B7 supertype allele). Peptides binding B*0702 with IC50 of ≦500 nM are identified using standard methods. These peptides are then tested for binding to other common B7-supertype molecules (e.g., B*3501, B*5101, B*5301, and B*5401). Peptides capable of binding to three or more of the five B7-supertype alleles tested are thereby identified.
Selection of A1 and A24 Motif-Bearing Epitopes
To further increase population coverage, HLA-A1 and -A24 epitopes can also be incorporated into vaccine compositions. An analysis of the 184P1E2 protein can also be performed to identify HLA-A1- and A24-motif-containing sequences.
High affinity and/or cross-reactive binding epitopes that bear other motif and/or supermotifs are identified using analogous methodology.
Example 14
Confirmation of Immunogenicity
Cross-reactive candidate CTL A2-supermotif-bearing peptides that are identified as described herein are selected to confirm in vitro immunogenicity. Confirmation is performed using the following methodology:
Target Cell Lines for Cellular Screening:
The 0.221A2.1 cell line, produced by transferring the HLA-A2.1 gene into the HLA-A, -B, -C null mutant human B-lymphoblastoid cell line 721.221, is used as the peptide-loaded target to measure activity of HLA-A2.1-restricted CTL. This cell line is grown in RPMI-1640 medium supplemented with antibiotics, sodium pyruvate, nonessential amino acids and 10% (v/v) heat inactivated FCS. Cells that express an antigen of interest, or transfectants comprising the gene encoding the antigen of interest, can be used as target cells to confirm the ability of peptide-specific CTLs to recognize endogenous antigen.
Primary CTL Induction Cultures:
Generation of Dendritic Cells (DC): PBMCs are thawed in RPMI with 30 μg/ml DNAse, washed twice and resuspended in complete medium (RPMI-1640 plus 5% AB human serum, non-essential amino acids, sodium pyruvate, L-glutamine and penicillin/streptomycin). The monocytes are purified by plating 10×106 PBMC/well in a 6-well plate. After 2 hours at 37° C., the non-adherent cells are removed by gently shaking the plates and aspirating the supernatants. The wells are washed a total of three times with 3 ml RPMI to remove most of the non-adherent and loosely adherent cells. Three ml of complete medium containing 50 ng/ml of GM-CSF and 1,000 U/ml of IL-4 are then added to each well. TNFα is added to the DCs on day 6 at 75 ng/ml and the cells are used for CTL induction cultures on day 7.
Induction of CTL with DC and Peptide: CD8+ T-cells are isolated by positive selection with Dynal immunomagnetic beads (Dynabeads® M-450) and the Detacha-Bead® reagent. Typically about 200-250×106 PBMC are processed to obtain 24×106 CD8+ T-cells (enough for a 48-well plate culture). Briefly, the PBMCs are thawed in RPMI with 30 μg/ml DNAse, washed once with PBS containing 1% human AB serum and resuspended in PBS/1% AB serum at a concentration of 20×106 cells/ml. The magnetic beads are washed 3 times with PBS/AB serum, added to the cells (140 μl beads/20×106 cells) and incubated for 1 hour at 4° C. with continuous mixing. The beads and cells are washed 4× with PBS/AB serum to remove the nonadherent cells and resuspended at 100×106 cells/ml (based on the original cell number) in PBS/AB serum containing 100 μl/ml Detacha-Bead® reagent and 30 μg/ml DNAse. The mixture is incubated for 1 hour at room temperature with continuous mixing. The beads are washed again with PBS/AB/DNAse to collect the CD8+ T-cells. The DC are collected and centrifuged at 1300 rpm for 5-7 minutes, washed once with PBS with 1% BSA, counted and pulsed with 40 μg/ml of peptide at a cell concentration of 1-2×106/ml in the presence of 3 μg/ml 12-microglobulin for 4 hours at 20° C. The DC are then irradiated (4,200 rads), washed 1 time with medium and counted again.
Setting up induction cultures: 0.25 ml cytokine-generated DC (at 1×105 cells/ml) are co-cultured with 0.25 ml of CD8+ T-cells (at 2×106 cell/ml) in each well of a 48-well plate in the presence of 10 ng/ml of IL-7. Recombinant human IL-10 is added the next day at a final concentration of 10 ng/ml and rhuman IL-2 is added 48 hours later at 10 IU/ml.
Restimulation of the induction cultures with peptide-pulsed adherent cells: Seven and fourteen days after the primary induction, the cells are restimulated with peptide-pulsed adherent cells. The PBMCs are thawed and washed twice with RPMI and DNAse. The cells are resuspended at 5×106 cells/ml and irradiated at  ̃4200 rads. The PBMCs are plated at 2×106 in 0.5 ml complete medium per well and incubated for 2 hours at 37° C. The plates are washed twice with RPMI by tapping the plate gently to remove the nonadherent cells and the adherent cells pulsed with 10 μg/ml of peptide in the presence of 3 μg/ml β2 microglobulin in 0.25 ml RPMI/5% AB per well for 2 hours at 37° C. Peptide solution from each well is aspirated and the wells are washed once with RPMI. Most of the media is aspirated from the induction cultures (CD8+ cells) and brought to 0.5 ml with fresh media. The cells are then transferred to the wells containing the peptide-pulsed adherent cells. Twenty four hours later recombinant human IL-10 is added at a final concentration of 10 ng/ml and recombinant human IL2 is added the next day and again 2-3 days later at 50IU/ml (Tsai et al., Critical Reviews in Immunology 18(1-2):65-75, 1998). Seven days later, the cultures are assayed for CTL activity in a 51Cr release assay. In some experiments the cultures are assayed for peptide-specific recognition in the in situ IFNγ ELISA at the time of the second restimulation followed by assay of endogenous recognition 7 days later. After expansion, activity is measured in both assays for a side-by-side comparison.
Measurement of CTL Lytic Activity by 51Cr Release.
Seven days after the second restimulation, cytotoxicity is determined in a standard (5 hr) 51Cr release assay by assaying individual wells at a single E:T. Peptide-pulsed targets are prepared by incubating the cells with 10 μg/ml peptide overnight at 37° C.
Adherent target cells are removed from culture flasks with trypsin-EDTA. Target cells are labeled with 200 μCi of 51Cr sodium chromate (Dupont, Wilmington, Del.) for 1 hour at 37° C. Labeled target cells are resuspended at 106 per ml and diluted 1:10 with K562 cells at a concentration of 3.3×106/ml (an NK-sensitive erythroblastoma cell line used to reduce non-specific lysis). Target cells (100 μl) and effectors (100 μl) are plated in 96 well round-bottom plates and incubated for 5 hours at 37° C. At that time, 100 μl of supernatant are collected from each well and percent lysis is determined according to the formula:

[(cpm of the test sample−cpm of the spontaneous 51Cr release sample)/(cpm of the maximal 51Cr release sample−cpm of the spontaneous 51Cr release sample)]×100.

Maximum and spontaneous release are determined by incubating the labeled targets with 1% Triton X-100 and media alone, respectively. A positive culture is defined as one in which the specific lysis (sample-background) is 10% or higher in the case of individual wells and is 15% or more at the two highest E:T ratios when expanded cultures are assayed.
In situ Measurement of Human IFNγ Production as an Indicator of Peptide-Specific and Endogenous Recognition
Immulon 2 plates are coated with mouse anti-human IFNγ monoclonal antibody (4 μg/ml 0.1M NaHCO3, pH8.2) overnight at 4° C. The plates are washed with Ca2+, Mg2+-free PBS/0.05% Tween 20 and blocked with PBS/10% FCS for two hours, after which the CTLs (100 μl/well) and targets (100 μl/well) are added to each well, leaving empty wells for the standards and blanks (which received media only). The target cells, either peptide-pulsed or endogenous targets, are used at a concentration of 1×106 cells/ml. The plates are incubated for 48 hours at 37° C. with 5% CO2.
Recombinant human IFN-gamma is added to the standard wells starting at 400 pg or 1200 pg/100 microliter/well and the plate incubated for two hours at 37° C. The plates are washed and 100 μl of biotinylated mouse anti-human IFN-gamma monoclonal antibody (2 microgram/ml in PBS/3% FCS/0.05% Tween 20) are added and incubated for 2 hours at room temperature. After washing again, 100 microliter HRP-streptavidin (1:4000) are added and the plates incubated for one hour at room temperature. The plates are then washed 6× with wash buffer, 100 microliter/well developing solution (TMB 1:1) are added, and the plates allowed to develop for 5-15 minutes. The reaction is stopped with 50 microliter/well 1M H3PO4 and read at OD450. A culture is considered positive if it measured at least 50 pg of IFN-gamma/well above background and is twice the background level of expression.
CTL Expansion.
Those cultures that demonstrate specific lytic activity against peptide-pulsed targets and/or tumor targets are expanded over a two week period with anti-CD3. Briefly, 5×104 CD8+ cells are added to a T25 flask containing the following: 1×106 irradiated (4,200 rad) PBMC (autologous or allogeneic) per ml, 2×105 irradiated (8,000 rad) EBV-transformed cells per ml, and OKT3 (anti-CD3) at 30 ng per ml in RPMI-1640 containing 10% (v/v) human AB serum, non-essential amino acids, sodium pyruvate, 25 μM 2-mercaptoethanol, L-glutamine and penicillin/streptomycin. Recombinant human IL2 is added 24 hours later at a final concentration of 2001 U/ml and every three days thereafter with fresh media at 50IU/ml. The cells are split if the cell concentration exceeds 1×106/ml and the cultures are assayed between days 13 and 15 at E:T ratios of 30, 10, 3 and 1:1 in the 51Cr release assay or at 1×106/ml in the in situ IFNγ assay using the same targets as before the expansion.
Cultures are expanded in the absence of anti-CD3+ as follows. Those cultures that demonstrate specific lytic activity against peptide and endogenous targets are selected and 5×104 CD8+ cells are added to a T25 flask containing the following: 1×106 autologous PBMC per ml which have been peptide-pulsed with 10 μg/ml peptide for two hours at 37° C. and irradiated (4,200 rad); 2×105 irradiated (8,000 rad) EBV-transformed cells per ml RPMI-1640 containing 10% (v/v) human AB serum, non-essential AA, sodium pyruvate, 25 mM 2-ME, L-glutamine and gentamicin.
Immunogenicity of A2 Supermotif-Bearing Peptides
A2-supermotif cross-reactive binding peptides are tested in the cellular assay for the ability to induce peptide-specific CTL in normal individuals. In this analysis, a peptide is typically considered to be an epitope if it induces peptide-specific CTLs in at least individuals, and preferably, also recognizes the endogenously expressed peptide.
Immunogenicity can also be confirmed using PBMCs isolated from patients bearing a tumor that expresses 184P1E2. Briefly, PBMCs are isolated from patients, re-stimulated with peptide-pulsed monocytes and assayed for the ability to recognize peptide-pulsed target cells as well as transfected cells endogenously expressing the antigen.
Evaluation of A*03/A11 Immunogenicity
HLA-A3 supermotif-bearing cross-reactive binding peptides are also evaluated for immunogenicity using methodology analogous for that used to evaluate the immunogenicity of the HLA-A2 supermotif peptides.
Evaluation of B7 Immunogenicity
Immunogenicity screening of the B7-supertype cross-reactive binding peptides identified as set forth herein are confirmed in a manner analogous to the confirmation of A2- and A3-supermotif-bearing peptides.
Peptides bearing other supermotifs/motifs, e.g., HLA-A1, HLA-A24 etc. are also confirmed using similar methodology
Example 15
Implementation of the Extended Supermotif to Improve the Binding Capacity of Native Epitopes by Creating Analogs
HLA motifs and supermotifs (comprising primary and/or secondary residues) are useful in the identification and preparation of highly cross-reactive native peptides, as demonstrated herein. Moreover, the definition of HLA motifs and supermotifs also allows one to engineer highly cross-reactive epitopes by identifying residues within a native peptide sequence which can be analoged to confer upon the peptide certain characteristics, e.g. greater cross-reactivity within the group of HLA molecules that comprise a supertype, and/or greater binding affinity for some or all of those HLA molecules. Examples of analoging peptides to exhibit modulated binding affinity are set forth in this example.
Analoging at Primary Anchor Residues
Peptide engineering strategies are implemented to further increase the cross-reactivity of the epitopes. For example, the main anchors of A2-supermotif-bearing peptides are altered, for example, to introduce a preferred L, I, V, or M at position 2, and I or V at the C-terminus.
To analyze the cross-reactivity of the analog peptides, each engineered analog is initially tested for binding to the prototype A2 supertype allele A*0201, then, if A*0201 binding capacity is maintained, for A2-supertype cross-reactivity.
Alternatively, a peptide is confirmed as binding one or all supertype members and then analoged to modulate binding affinity to any one (or more) of the supertype members to add population coverage.
The selection of analogs for immunogenicity in a cellular screening analysis is typically further restricted by the capacity of the parent wild type (WT) peptide to bind at least weakly, i.e., bind at an IC500f 5000 nM or less, to three of more A2 supertype alleles. The rationale for this requirement is that the WT peptides must be present endogenously in sufficient quantity to be biologically relevant. Analoged peptides have been shown to have increased immunogenicity and cross-reactivity by T cells specific for the parent epitope (see, e.g., Parkhurst et al., J. Immunol. 157:2539, 1996; and Pogue et al., Proc. Natl. Acad. Sci. USA 92:8166, 1995).
In the cellular screening of these peptide analogs, it is important to confirm that analog-specific CTLs are also able to recognize the wild-type peptide and, when possible, target cells that endogenously express the epitope.
Analoging of HLA-A3 and B7-Supermotif-Bearing Peptides
Analogs of HLA-A3 supermotif-bearing epitopes are generated using strategies similar to those employed in analoging HLA-A2 supermotif-bearing peptides. For example, peptides binding to 3/5 of the A3-supertype molecules are engineered at primary anchor residues to possess a preferred residue (V, S, M, or A) at position 2.
The analog peptides are then tested for the ability to bind A*03 and A*11 (prototype A3 supertype alleles). Those peptides that demonstrate ≦500 nM binding capacity are then confirmed as having A3-supertype cross-reactivity.
Similarly to the A2- and A3-motif bearing peptides, peptides binding 3 or more B7-supertype alleles can be improved, where possible, to achieve increased cross-reactive binding or greater binding affinity or binding half life. B7 supermotif-bearing peptides are, for example, engineered to possess a preferred residue (V, I, L, or F) at the C-terminal primary anchor position, as demonstrated by Sidney et al. (J. Immunol. 157:3480-3490, 1996).
Analoging at primary anchor residues of other motif and/or supermotif-bearing epitopes is performed in a like manner.
The analog peptides are then be confirmed for immunogenicity, typically in a cellular screening assay. Again, it is generally important to demonstrate that analog-specific CTLs are also able to recognize the wild-type peptide and, when possible, targets that endogenously express the epitope.
Analoging at Secondary Anchor Residues
Moreover, HLA supermotifs are of value in engineering highly cross-reactive peptides and/or peptides that bind HLA molecules with increased affinity by identifying particular residues at secondary anchor positions that are associated with such properties. For example, the binding capacity of a B7 supermotif-bearing peptide with an F residue at position 1 is analyzed. The peptide is then analoged to, for example, substitute L for F at position 1. The analoged peptide is evaluated for increased binding affinity, binding half life and/or increased cross-reactivity. Such a procedure identifies analoged peptides with enhanced properties.
Engineered analogs with sufficiently improved binding capacity or cross-reactivity can also be tested for immunogenicity in HLA-B7-transgenic mice, following for example, IFA immunization or lipopeptide immunization. Analoged peptides are additionally tested for the ability to stimulate a recall response using PBMC from patients with 184P1E2-expressing tumors.
Other Analoging Strategies
Another form of peptide analoging, unrelated to anchor positions, involves the substitution of a cysteine with α-amino butyric acid. Due to its chemical nature, cysteine has the propensity to form disulfide bridges and sufficiently alter the peptide structurally so as to reduce binding capacity. Substitution of α-amino butyric acid for cysteine not only alleviates this problem, but has been shown to improve binding and crossbinding capabilities in some instances (see, e.g., the review by Sette et al., In: Persistent Viral Infections, Eds. R. Ahmed and I. Chen, John Wiley & Sons, England, 1999).
Thus, by the use of single amino acid substitutions, the binding properties and/or cross-reactivity of peptide ligands for HLA supertype molecules can be modulated.
Example 16
Identification and Confirmation of 184P1E2-Derived Sequences with HLA-DR Binding Motifs
Peptide epitopes bearing an HLA class II supermotif or motif are identified and confirmed as outlined below using methodology similar to that described for HLA Class I peptides.
Selection of HLA-DR-Supermotif-Bearing Epitopes.
To identify 184P1E2-derived, HLA class II HTL epitopes, a 184P1E2 antigen is analyzed for the presence of sequences bearing an HLA-DR-motif or supermotif. Specifically, 15-mer sequences are selected comprising a DR-supermotif, comprising a 9-mer core, and three-residue N- and C-terminal flanking regions (15 amino acids total).
Protocols for predicting peptide binding to DR molecules have been developed (Southwood et al., J. Immunol. 160:3363-3373, 1998). These protocols, specific for individual DR molecules, allow the scoring, and ranking, of 9-mer core regions. Each protocol not only scores peptide sequences for the presence of DR-supermotif primary anchors (i.e., at position 1 and position 6) within a 9-mer core, but additionally evaluates sequences for the presence of secondary anchors. Using allele-specific selection tables (see, e.g., Southwood et al., ibid.), it has been found that these protocols efficiently select peptide sequences with a high probability of binding a particular DR molecule. Additionally, it has been found that performing these protocols in tandem, specifically those for DR1, DR4w4, and DR7, can efficiently select DR cross-reactive peptides.
The 184P1E2-derived peptides identified above are tested for their binding capacity for various common HLA-DR molecules. All peptides are initially tested for binding to the DR molecules in the primary panel: DR1, DR4w4, and DR7. Peptides binding at least two of these three DR molecules are then tested for binding to DR2w2 β1, DR2w2 β2, DR6w19, and DR9 molecules in secondary assays. Finally, peptides binding at least two of the four secondary panel DR molecules, and thus cumulatively at least four of seven different DR molecules, are screened for binding to DR4w15, DR5w11, and DR8w2 molecules in tertiary assays. Peptides binding at least seven of the ten DR molecules comprising the primary, secondary, and tertiary screening assays are considered cross-reactive DR binders. 184P1E2-derived peptides found to bind common HLA-DR alleles are of particular interest.
Selection of DR3 Motif Peptides
Because HLA-DR3 is an allele that is prevalent in Caucasian, Black, and Hispanic populations, DR3 binding capacity is a relevant criterion in the selection of HTL epitopes. Thus, peptides shown to be candidates may also be assayed for their DR3 binding capacity. However, in view of the binding specificity of the DR3 motif, peptides binding only to DR3 can also be considered as candidates for inclusion in a vaccine formulation.
To efficiently identify peptides that bind DR3, target 184P1E2 antigens are analyzed for sequences carrying one of the two DR3-specific binding motifs reported by Geluk et al. (J. Immunol. 152:5742-5748, 1994). The corresponding peptides are then synthesized and confirmed as having the ability to bind DR3 with an affinity of 1 μM or better, i.e., less than 1 μM. Peptides are found that meet this binding criterion and qualify as HLA class II high affinity binders.
DR3 binding epitopes identified in this manner are included in vaccine compositions with DR supermotif-bearing peptide epitopes.
Similarly to the case of HLA class I motif-bearing peptides, the class II motif-bearing peptides are analoged to improve affinity or cross-reactivity. For example, aspartic acid at position 4 of the 9-mer core sequence is an optimal residue for DR3 binding, and substitution for that residue often improves DR 3 binding.
Example 17
Immunogenicity of 184P1E2-Derived HTL Epitopes
This example determines immunogenic DR supermotif- and DR3 motif-bearing epitopes among those identified using the methodology set forth herein.
Immunogenicity of HTL epitopes are confirmed in a manner analogous to the determination of immunogenicity of CTL epitopes, by assessing the ability to stimulate HTL responses and/or by using appropriate transgenic mouse models. Immunogenicity is determined by screening for: 1.) in vitro primary induction using normal PBMC or 2.) recall responses from patients who have 184P1E2-expressing tumors.
Example 18
Calculation of Phenotypic Frequencies of HLA-Supertypes in Various Ethnic Backgrounds to Determine Breadth of Population Coverage
This example illustrates the assessment of the breadth of population coverage of a vaccine composition comprised of multiple epitopes comprising multiple supermotifs and/or motifs.
In order to analyze population coverage, gene frequencies of HLA alleles are determined. Gene frequencies for each HLA allele are calculated from antigen or allele frequencies utilizing the binomial distribution formulae gf=1−(SQRT(1−af)) (see, e.g., Sidney et al., Human Immunol 45:79-93, 1996). To obtain overall phenotypic frequencies, cumulative gene frequencies are calculated, and the cumulative antigen frequencies derived by the use of the inverse formula [af=1−(1−Cgf)2].
Where frequency data is not available at the level of DNA typing, correspondence to the serologically defined antigen frequencies is assumed. To obtain total potential supertype population coverage no linkage disequilibrium is assumed, and only alleles confirmed to belong to each of the supertypes are included (minimal estimates). Estimates of total potential coverage achieved by inter-loci combinations are made by adding to the A coverage the proportion of the non-A covered population that could be expected to be covered by the B alleles considered (e.g., total=A+B*(1−A)). Confirmed members of the A3-like supertype are A3, All, A31, A*3301, and A*6801. Although the A3-like supertype may also include A34, A66, and A*7401, these alleles were not included in overall frequency calculations. Likewise, confirmed members of the A2-like supertype family are A*0201, A*0202, A*0203, A*0204, A*0205, A*0206, A*0207, A*6802, and A*6901. Finally, the B7-like supertype-confirmed alleles are: B7, B*3501-03, B51, B*5301, B*5401, B*5501-2, B*5601, B*6701, and B*7801 (potentially also B*1401, B*3504-06, B*4201, and B*5602).
Population coverage achieved by combining the A2-, A3- and B7-supertypes is approximately 86% in five major ethnic groups. Coverage may be extended by including peptides bearing the A1 and A24 motifs. On average, A1 is present in 12% and A24 in 29% of the population across five different major ethnic groups (Caucasian, North American Black, Chinese, Japanese, and Hispanic). Together, these alleles are represented with an average frequency of 39% in these same ethnic populations. The total coverage across the major ethnicities when A1 and A24 are combined with the coverage of the A2-, A3- and B7-supertype alleles is >95%. An analogous approach can be used to estimate population coverage achieved with combinations of class II motif-bearing epitopes.
Immunogenicity studies in humans (e.g., Bertoni et al., J. Clin. Invest. 100:503, 1997; Doolan et al., Immunity 7:97, 1997; and Threlkeld et al., J. Immunol. 159:1648, 1997) have shown that highly cross-reactive binding peptides are almost always recognized as epitopes. The use of highly cross-reactive binding peptides is an important selection criterion in identifying candidate epitopes for inclusion in a vaccine that is immunogenic in a diverse population.
With a sufficient number of epitopes (as disclosed herein and from the art), an average population coverage is predicted to be greater than 95% in each of five major ethnic populations. The game theory Monte Carlo simulation analysis, which is known in the art (see e.g., Osborne, M. J. and Rubinstein, A. “A course in game theory” MIT Press, 1994), can be used to estimate what percentage of the individuals in a population comprised of the Caucasian, North American Black, Japanese, Chinese, and Hispanic ethnic groups would recognize the vaccine epitopes described herein. A preferred percentage is 90%. A more preferred percentage is 95%.
Example 19
CTL Recognition of Endogenously Processed Antigens after Priming
This example confirms that CTL induced by native or analoged peptide epitopes identified and selected as described herein recognize endogenously synthesized, i.e., native antigens.
Effector cells isolated from transgenic mice that are immunized with peptide epitopes, for example HLA-A2 supermotif-bearing epitopes, are re-stimulated in vitro using peptide-coated stimulator cells. Six days later, effector cells are assayed for cytotoxicity and the cell lines that contain peptide-specific cytotoxic activity are further re-stimulated. An additional six days later, these cell lines are tested for cytotoxic activity on 51Cr labeled Jurkat-A2.1/Kb target cells in the absence or presence of peptide, and also tested on 51Cr labeled target cells bearing the endogenously synthesized antigen, i.e. cells that are stably transfected with 184P1E2 expression vectors.
The results demonstrate that CTL lines obtained from animals primed with peptide epitope recognize endogenously synthesized 184P1E2 antigen. The choice of transgenic mouse model to be used for such an analysis depends upon the epitope(s) that are being evaluated. In addition to HLA-A*0201/Kb transgenic mice, several other transgenic mouse models including mice with human A11, which may also be used to evaluate A3 epitopes, and B7 alleles have been characterized and others (e.g., transgenic mice for HLA-A1 and A24) are being developed. HLA-DR1 and HLA-DR3 mouse models have also been developed, which may be used to evaluate HTL epitopes.
Example 20
Activity of CTL-HTL Conjugated Epitopes in Transgenic Mice
This example illustrates the induction of CTLs and HTLs in transgenic mice, by use of a 184P1E2-derived CTL and HTL peptide vaccine compositions. The vaccine composition used herein comprise peptides to be administered to a patient with a 184P1E2-expressing tumor. The peptide composition can comprise multiple CTL and/or HTL epitopes. The epitopes are identified using methodology as described herein. This example also illustrates that enhanced immunogenicity can be achieved by inclusion of one or more HTL epitopes in a CTL vaccine composition; such a peptide composition can comprise an HTL epitope conjugated to a CTL epitope. The CTL epitope can be one that binds to multiple HLA family members at an affinity of 500 nM or less, or analogs of that epitope. The peptides may be lipidated, if desired.
Immunization procedures: Immunization of transgenic mice is performed as described (Alexander et al., J. Immunol. 159:4753-4761, 1997). For example, A2/Kb mice, which are transgenic for the human HLA A2.1 allele and are used to confirm the immunogenicity of HLA-A*0201 motif- or HLA-A2 supermotif-bearing epitopes, and are primed subcutaneously (base of the tail) with a 0.1 ml of peptide in Incomplete Freund's Adjuvant, or if the peptide composition is a lipidated CTL/HTL conjugate, in DMSO/saline, or if the peptide composition is a polypeptide, in PBS or Incomplete Freund's Adjuvant. Seven days after priming, splenocytes obtained from these animals are restimulated with syngenic irradiated LPS-activated lymphoblasts coated with peptide.
Cell lines: Target cells for peptide-specific cytotoxicity assays are Jurkat cells transfected with the HLA-A2.1/Kb chimeric gene (e.g., Vitiello et al., J. Exp. Med. 173:1007, 1991)
In vitro CTL activation: One week after priming, spleen cells (30×106 cells/flask) are co-cultured at 37° C. with syngeneic, irradiated (3000 rads), peptide coated lymphoblasts (10×106 cells/flask) in 10 ml of culture medium/T25 flask. After six days, effector cells are harvested and assayed for cytotoxic activity.
Assay for cytotoxic activity: Target cells (1.0 to 1.5×106) are incubated at 37° C. in the presence of 200 μl of 51Cr. After 60 minutes, cells are washed three times and resuspended in R10 medium. Peptide is added where required at a concentration of 1 μg/ml. For the assay, 104 51Cr-labeled target cells are added to different concentrations of effector cells (final volume of 200 μl) in U-bottom 96-well plates. After a six hour incubation period at 37° C., a 0.1 ml aliquot of supernatant is removed from each well and radioactivity is determined in a Micromedic automatic gamma counter. The percent specific lysis is determined by the formula: percent specific release=100× (experimental release−spontaneous release)/(maximum release−spontaneous release). To facilitate comparison between separate CTL assays run under the same conditions, % 51Cr release data is expressed as lytic units/106 cells. One lytic unit is arbitrarily defined as the number of effector cells required to achieve 30% lysis of 10,000 target cells in a six hour 51Cr release assay. To obtain specific lytic units/106, the lytic units/106 obtained in the absence of peptide is subtracted from the lytic units/106 obtained in the presence of peptide. For example, if 30% 51Cr release is obtained at the effector (E): target (T) ratio of 50:1 (i.e., 5×105 effector cells for 10,000 targets) in the absence of peptide and 5:1 (i.e., 5×104 effector cells for 10,000 targets) in the presence of peptide, the specific lytic units would be: [(1/50,000)−(1/500,000)]×106=18 LU.
The results are analyzed to assess the magnitude of the CTL responses of animals injected with the immunogenic CTL/HTL conjugate vaccine preparation and are compared to the magnitude of the CTL response achieved using, for example, CTL epitopes as outlined above in the Example entitled “Confirmation of Immunogenicity.” Analyses similar to this may be performed to confirm the immunogenicity of peptide conjugates containing multiple CTL epitopes and/or multiple HTL epitopes. In accordance with these procedures, it is found that a CTL response is induced, and concomitantly that an HTL response is induced upon administration of such compositions.
Example 21
Selection of CTL and HTL Epitopes for Inclusion in a 184P1E2-Specific Vaccine
This example illustrates a procedure for selecting peptide epitopes for vaccine compositions of the invention. The peptides in the composition can be in the form of a nucleic acid sequence, either single or one or more sequences (i.e., minigene) that encodes peptide(s), or can be single and/or polyepitopic peptides.
The following principles are utilized when selecting a plurality of epitopes for inclusion in a vaccine composition. Each of the following principles is balanced in order to make the selection.
Epitopes are selected which, upon administration, mimic immune responses that are correlated with 184P1E2 clearance. The number of epitopes used depends on observations of patients who spontaneously clear 184P1E2. For example, if it has been observed that patients who spontaneously clear 184P1E2-expressing cells generate an immune response to at least three (3) epitopes from 184P1E2 antigen, then at least three epitopes should be included for HLA class I. A similar rationale is used to determine HLA class II epitopes.
Epitopes are often selected that have a binding affinity of an IC50 of 500 nM or less for an HLA class I molecule, or for class II, an IC50 of 1000 nM or less; or HLA Class I peptides with high binding scores from the BIMAS web site, at URL bimas.dcrt.nih.gov/.
In order to achieve broad coverage of the vaccine through out a diverse population, sufficient supermotif bearing peptides, or a sufficient array of allele-specific motif bearing peptides, are selected to give broad population coverage. In one embodiment, epitopes are selected to provide at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess breadth, or redundancy, of population coverage.
When creating polyepitopic compositions, or a minigene that encodes same, it is typically desirable to generate the smallest peptide possible that encompasses the epitopes of interest. The principles employed are similar, if not the same, as those employed when selecting a peptide comprising nested epitopes. For example, a protein sequence for the vaccine composition is selected because it has maximal number of epitopes contained within the sequence, i.e., it has a high concentration of epitopes. Epitopes may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Each epitope can be exposed and bound by an HLA molecule upon administration of such a peptide. A multi-epitopic, peptide can be generated synthetically, recombinantly, or via cleavage from the native source. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties of the polyepitopic peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes. This embodiment provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally such an embodiment provides for the possibility of motif-bearing epitopes for an HLA makeup that is presently unknown. Furthermore, this embodiment (absent the creating of any analogs) directs the immune response to multiple peptide sequences that are actually present in 184P1E2, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing nucleic acid vaccine compositions. Related to this embodiment, computer programs can be derived in accordance with principles in the art, which identify in a target sequence, the greatest number of epitopes per sequence length.
A vaccine composition comprised of selected peptides, when administered, is safe, efficacious, and elicits an immune response similar in magnitude to an immune response that controls or clears cells that bear or overexpress 184P1E2.
Example 22
Construction of “Minigene” Multi-Epitope DNA Plasmids
This example discusses the construction of a minigene expression plasmid. Minigene plasmids may, of course, contain various configurations of B cell, CTL and/or HTL epitopes or epitope analogs as described herein.
A minigene expression plasmid typically includes multiple CTL and HTL peptide epitopes. In the present example, HLA-A2, -A3, -B7 supermotif-bearing peptide epitopes and HLA-A1 and -A24 motif-bearing peptide epitopes are used in conjunction with DR supermotif-bearing epitopes and/or DR3 epitopes. HLA class I supermotif or motif-bearing peptide epitopes derived 184P1E2, are selected such that multiple supermotifs/motifs are represented to ensure broad population coverage. Similarly, HLA class II epitopes are selected from 184P1E2 to provide broad population coverage, i.e. both HLA DR-1-4-7 supermotif-bearing epitopes and HLA DR-3 motif-bearing epitopes are selected for inclusion in the minigene construct. The selected CTL and HTL epitopes are then incorporated into a minigene for expression in an expression vector.
Such a construct may additionally include sequences that direct the HTL epitopes to the endoplasmic reticulum. For example, the Ii protein may be fused to one or more HTL epitopes as described in the art, wherein the CLIP sequence of the Ii protein is removed and replaced with an HLA class II epitope sequence so that HLA class II epitope is directed to the endoplasmic reticulum, where the epitope binds to an HLA class II molecules.
This example illustrates the methods to be used for construction of a minigene-bearing expression plasmid. Other expression vectors that may be used for minigene compositions are available and known to those of skill in the art.
The minigene DNA plasmid of this example contains a consensus Kozak sequence and a consensus murine kappa Ig-light chain signal sequence followed by CTL and/or HTL epitopes selected in accordance with principles disclosed herein. The sequence encodes an open reading frame fused to the Myc and His antibody epitope tag coded for by the pcDNA 3.1 Myc-His vector.
Overlapping oligonucleotides that can, for example, average about 70 nucleotides in length with 15 nucleotide overlaps, are synthesized and HPLC-purified. The oligonucleotides encode the selected peptide epitopes as well as appropriate linker nucleotides, Kozak sequence, and signal sequence. The final multiepitope minigene is assembled by extending the overlapping oligonucleotides in three sets of reactions using PCR. A Perkin/Elmer 9600 PCR machine is used and a total of 30 cycles are performed using the following conditions: 95° C. for 15 sec, annealing temperature (5° below the lowest calculated Tm of each primer pair) for 30 sec, and 72° C. for 1 min.
For example, a minigene is prepared as follows. For a first PCR reaction, 5 μg of each of two oligonucleotides are annealed and extended: In an example using eight oligonucleotides, i.e., four pairs of primers, oligonucleotides 1+2, 3+4, 5+6, and 7+8 are combined in 100 μl reactions containing Pfu polymerase buffer (1×=10 mM KCL, 10 mM (NH4)2SO4, 20 mM Tris-chloride, pH 8.75, 2 mM MgSO4, 0.1% Triton X-100, 100 μg/ml BSA), 0.25 mM each dNTP, and 2.5 U of Pfu polymerase. The full-length dimer products are gel-purified, and two reactions containing the product of 1+2 and 3+4, and the product of 5+6 and 7+8 are mixed, annealed, and extended for 10 cycles. Half of the two reactions are then mixed, and 5 cycles of annealing and extension carried out before flanking primers are added to amplify the full length product. The full-length product is gel-purified and cloned into pCR-blunt (Invitrogen) and individual clones are screened by sequencing.
Example 23
The Plasmid Construct and the Degree to which it Induces Immunogenicity
The degree to which a plasmid construct, for example a plasmid constructed in accordance with the previous Example, is able to induce immunogenicity is confirmed in vitro by determining epitope presentation by APC following transduction or transfection of the APC with an epitope-expressing nucleic acid construct. Such a study determines “antigenicity” and allows the use of human APC. The assay determines the ability of the epitope to be presented by the APC in a context that is recognized by a T cell by quantifying the density of epitope-HLA class I complexes on the cell surface. Quantitation can be performed by directly measuring the amount of peptide eluted from the APC (see, e.g., Sijts et al., J. Immunol. 156:683-692, 1996; Demotz et al., Nature 342:682-684, 1989); or the number of peptide-HLA class I complexes can be estimated by measuring the amount of lysis or lymphokine release induced by diseased or transfected target cells, and then determining the concentration of peptide necessary to obtain equivalent levels of lysis or lymphokine release (see, e.g., Kageyama et al., J. Immunol. 154:567-576, 1995).
Alternatively, immunogenicity is confirmed through in vivo injections into mice and subsequent in vitro assessment of CTL and HTL activity, which are analyzed using cytotoxicity and proliferation assays, respectively, as detailed e.g., in Alexander et al., Immunity 1:751-761, 1994.
For example, to confirm the capacity of a DNA minigene construct containing at least one HLA-A2 supermotif peptide to induce CTLs in vivo, HLA-A2.1/Kb transgenic mice, for example, are immunized intramuscularly with 100 μg of naked cDNA. As a means of comparing the level of CTLs induced by cDNA immunization, a control group of animals is also immunized with an actual peptide composition that comprises multiple epitopes synthesized as a single polypeptide as they would be encoded by the minigene.
Splenocytes from immunized animals are stimulated twice with each of the respective compositions (peptide epitopes encoded in the minigene or the polyepitopic peptide), then assayed for peptide-specific cytotoxic activity in a 51Cr release assay. The results indicate the magnitude of the CTL response directed against the A2-restricted epitope, thus indicating the in vivo immunogenicity of the minigene vaccine and polyepitopic vaccine.
It is, therefore, found that the minigene elicits immune responses directed toward the HLA-A2 supermotif peptide epitopes as does the polyepitopic peptide vaccine. A similar analysis is also performed using other HLA-A3 and HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 and HLA-B7 motif or supermotif epitopes, whereby it is also found that the minigene elicits appropriate immune responses directed toward the provided epitopes.
To confirm the capacity of a class II epitope-encoding minigene to induce HTLs in vivo, DR transgenic mice, or for those epitopes that cross react with the appropriate mouse MHC molecule, I-Ab-restricted mice, for example, are immunized intramuscularly with 100 μg of plasmid DNA. As a means of comparing the level of HTLs induced by DNA immunization, a group of control animals is also immunized with an actual peptide composition emulsified in complete Freund's adjuvant. CD4+ T cells, i.e. HTLs, are purified from splenocytes of immunized animals and stimulated with each of the respective compositions (peptides encoded in the minigene). The HTL response is measured using a 3H-thymidine incorporation proliferation assay, (see, e.g., Alexander et al. Immunity 1:751-761, 1994). The results indicate the magnitude of the HTL response, thus demonstrating the in vivo immunogenicity of the mini gene.
DNA minigenes, constructed as described in the previous Example, can also be confirmed as a vaccine in combination with a boosting agent using a prime boost protocol. The boosting agent can consist of recombinant protein (e.g., Barnett et al., Aids Res. and Human Retroviruses 14, Supplement 3:S299-S309, 1998) or recombinant vaccinia, for example, expressing a minigene or DNA encoding the complete protein of interest (see, e.g., Hanke et al., Vaccine 16:439-445, 1998; Sedegah et al., Proc. Natl. Acad. Sci. USA 95:7648-53, 1998; Hanke and McMichael, Immunol. Letters 66:177-181, 1999; and Robinson et al., Nature Med. 5:526-34, 1999).
For example, the efficacy of the DNA minigene used in a prime boost protocol is initially evaluated in transgenic mice. In this example, A2.1/Kb transgenic mice are immunized IM with 100 μg of a DNA minigene encoding the immunogenic peptides including at least one HLA-A2 supermotif-bearing peptide. After an incubation period (ranging from 3-9 weeks), the mice are boosted IP with 107 pfu/mouse of a recombinant vaccinia virus expressing the same sequence encoded by the DNA minigene. Control mice are immunized with 100 μg of DNA or recombinant vaccinia without the minigene sequence, or with DNA encoding the minigene, but without the vaccinia boost. After an additional incubation period of two weeks, splenocytes from the mice are immediately assayed for peptide-specific activity in an ELISPOT assay. Additionally, splenocytes are stimulated in vitro with the A2-restricted peptide epitopes encoded in the minigene and recombinant vaccinia, then assayed for peptide-specific activity in an alpha, beta and/or gamma IFN ELISA.
It is found that the minigene utilized in a prime-boost protocol elicits greater immune responses toward the HLA-A2 supermotif peptides than with DNA alone. Such an analysis can also be performed using HLA-A11 or HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 or HLA-B7 motif or supermotif epitopes. The use of prime boost protocols in humans is described below in the Example entitled “Induction of CTL Responses Using a Prime Boost Protocol.”
Example 24
Peptide Compositions for Prophylactic Uses
Vaccine compositions of the present invention can be used to prevent 184P1E2 expression in persons who are at risk for tumors that bear this antigen. For example, a polyepitopic peptide epitope composition (or a nucleic acid comprising the same) containing multiple CTL and HTL epitopes such as those selected in the above Examples, which are also selected to target greater than 80% of the population, is administered to individuals at risk for a 184P1E2-associated tumor.
For example, a peptide-based composition is provided as a single polypeptide that encompasses multiple epitopes. The vaccine is typically administered in a physiological solution that comprises an adjuvant, such as Incomplete Freunds Adjuvant. The dose of peptide for the initial immunization is from about 1 to about 50,000 μg, generally 100-5,000 μg, for a 70 kg patient. The initial administration of vaccine is followed by booster dosages at 4 weeks followed by evaluation of the magnitude of the immune response in the patient, by techniques that determine the presence of epitope-specific CTL populations in a PBMC sample. Additional booster doses are administered as required. The composition is found to be both safe and efficacious as a prophylaxis against 184P1E2-associated disease.
Alternatively, a composition typically comprising transfecting agents is used for the administration of a nucleic acid-based vaccine in accordance with methodologies known in the art and disclosed herein.
Example 25
Polyepitopic Vaccine Compositions Derived from Native 184P1E2 Sequences
A native 184P1E2 polyprotein sequence is analyzed, preferably using computer algorithms defined for each class I and/or class II supermotif or motif, to identify “relatively short” regions of the polyprotein that comprise multiple epitopes. The “relatively short” regions are preferably less in length than an entire native antigen. This relatively short sequence that contains multiple distinct or overlapping, “nested” epitopes can be used to generate a minigene construct. The construct is engineered to express the peptide, which corresponds to the native protein sequence. The “relatively short” peptide is generally less than 250 amino acids in length, often less than 100 amino acids in length, preferably less than 75 amino acids in length, and more preferably less than 50 amino acids in length. The protein sequence of the vaccine composition is selected because it has maximal number of epitopes contained within the sequence, i.e., it has a high concentration of epitopes. As noted herein, epitope motifs may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes.
The vaccine composition will include, for example, multiple CTL epitopes from 184P1E2 antigen and at least one HTL epitope. This polyepitopic native sequence is administered either as a peptide or as a nucleic acid sequence which encodes the peptide. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties of the polyepitopic peptide.
The embodiment of this example provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally, such an embodiment provides for the possibility of motif-bearing epitopes for an HLA makeup(s) that is presently unknown. Furthermore, this embodiment (excluding an analoged embodiment) directs the immune response to multiple peptide sequences that are actually present in native 184P1E2, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing peptide or nucleic acid vaccine compositions.
Related to this embodiment, computer programs are available in the art which can be used to identify in a target sequence, the greatest number of epitopes per sequence length.
Example 26
Polyepitopic Vaccine Compositions from Multiple Antigens
The 184P1E2 peptide epitopes of the present invention are used in conjunction with epitopes from other target tumor-associated antigens, to create a vaccine composition that is useful for the prevention or treatment of cancer that expresses 184P1E2 and such other antigens. For example, a vaccine composition can be provided as a single polypeptide that incorporates multiple epitopes from 184P1E2 as well as tumor-associated antigens that are often expressed with a target cancer associated with 184P1E2 expression, or can be administered as a composition comprising a cocktail of one or more discrete epitopes. Alternatively, the vaccine can be administered as a minigene construct or as dendritic cells which have been loaded with the peptide epitopes in vitro.
Example 27
Use of Peptides to Evaluate an Immune Response
Peptides of the invention may be used to analyze an immune response for the presence of specific antibodies, CTL or HTL directed to 184P1E2. Such an analysis can be performed in a manner described by Ogg et al., Science 279:2103-2106, 1998. In this Example, peptides in accordance with the invention are used as a reagent for diagnostic or prognostic purposes, not as an immunogen.
In this example highly sensitive human leukocyte antigen tetrameric complexes (“tetramers”) are used for a cross-sectional analysis of, for example, 184P1E2 HLA-A*0201-specific CTL frequencies from HLA A*0201-positive individuals at different stages of disease or following immunization comprising a 184P1E2 peptide containing an A*0201 motif. Tetrameric complexes are synthesized as described (Musey et al., N. Engl. J. Med. 337:1267, 1997). Briefly, purified HLA heavy chain (A*0201 in this example) and β2-microglobulin are synthesized by means of a prokaryotic expression system. The heavy chain is modified by deletion of the transmembrane-cytosolic tail and COOH-terminal addition of a sequence containing a BirA enzymatic biotinylation site. The heavy chain, β2-microglobulin, and peptide are refolded by dilution. The 45-kD refolded product is isolated by fast protein liquid chromatography and then biotinylated by BirA in the presence of biotin (Sigma, St. Louis, Mo.), adenosine 5′ triphosphate and magnesium. Streptavidin-phycoerythrin conjugate is added in a 1:4 molar ratio, and the tetrameric product is concentrated to 1 mg/ml. The resulting product is referred to as tetramer-phycoerythrin.
For the analysis of patient blood samples, approximately one million PBMCs are centrifuged at 300 g for 5 minutes and resuspended in 50 μl of cold phosphate-buffered saline. Tri-color analysis is performed with the tetramer-phycoerythrin, along with anti-CD8-Tricolor, and anti-CD38. The PBMCs are incubated with tetramer and antibodies on ice for 30 to 60 min and then washed twice before formaldehyde fixation. Gates are applied to contain >99.98% of control samples. Controls for the tetramers include both A*0201-negative individuals and A*0201-positive non-diseased donors. The percentage of cells stained with the tetramer is then determined by flow cytometry. The results indicate the number of cells in the PBMC sample that contain epitope-restricted CTLs, thereby readily indicating the extent of immune response to the 184P1E2 epitope, and thus the status of exposure to 184P1E2, or exposure to a vaccine that elicits a protective or therapeutic response.
Example 28
Use of Peptide Epitopes to Evaluate Recall Responses
The peptide epitopes of the invention are used as reagents to evaluate T cell responses, such as acute or recall responses, in patients. Such an analysis may be performed on patients who have recovered from 184P1E2-associated disease or who have been vaccinated with a 184P1E2 vaccine.
For example, the class I restricted CTL response of persons who have been vaccinated may be analyzed. The vaccine may be any 184P1E2 vaccine. PBMC are collected from vaccinated individuals and HLA typed. Appropriate peptide epitopes of the invention that, optimally, bear supermotifs to provide cross-reactivity with multiple HLA supertype family members, are then used for analysis of samples derived from individuals who bear that HLA type.
PBMC from vaccinated individuals are separated on Ficoll-Histopaque density gradients (Sigma Chemical Co., St. Louis, Mo.), washed three times in HBSS (GIBCO Laboratories), resuspended in RPMI-1640 (GIBCO Laboratories) supplemented with L-glutamine (2 mM), penicillin (50 U/ml), streptomycin (50 μg/ml), and Hepes (10 mM) containing 10% heat-inactivated human AB serum (complete RPMI) and plated using microculture formats. A synthetic peptide comprising an epitope of the invention is added at 10 μg/ml to each well and HBV core 128-140 epitope is added at 1 μg/ml to each well as a source of T cell help during the first week of stimulation.
In the microculture format, 4×105 PBMC are stimulated with peptide in 8 replicate cultures in 96-well round bottom plate in 100 μl/well of complete RPMI. On days 3 and 10, 100 μl of complete RPMI and 20 U/ml final concentration of rIL-2 are added to each well. On day 7 the cultures are transferred into a 96-well flat-bottom plate and restimulated with peptide, rIL-2 and 105 irradiated (3,000 rad) autologous feeder cells. The cultures are tested for cytotoxic activity on day 14. A positive CTL response requires two or more of the eight replicate cultures to display greater than 10% specific 51Cr release, based on comparison with non-diseased control subjects as previously described (Rehermann, et al., Nature Med. 2:1104, 1108, 1996; Rehermann et al., J. Clin. Invest. 97:1655-1665, 1996; and Rehermann et al. J. Clin. Invest. 98:1432-1440, 1996).
Target cell lines are autologous and allogeneic EBV-transformed B-LCL that are either purchased from the American Society for Histocompatibility and Immunogenetics (ASHI, Boston, Mass.) or established from the pool of patients as described (Guilhot, et al. J. Virol. 66:2670-2678, 1992).
Cytotoxicity assays are performed in the following manner. Target cells consist of either allogeneic HLA-matched or autologous EBV-transformed B lymphoblastoid cell line that are incubated overnight with the synthetic peptide epitope of the invention at 10 μM, and labeled with 100 μCi of 51Cr (Amersham Corp., Arlington Heights, Ill.) for 1 hour after which they are washed four times with HBSS.
Cytolytic activity is determined in a standard 4-h, split well 51Cr release assay using U-bottomed 96 well plates containing 3,000 targets/well. Stimulated PBMC are tested at effector/target (E/T) ratios of 20-50:1 on day 14. Percent cytotoxicity is determined from the formula: 100×[(experimental release-spontaneous release)/maximum release-spontaneous release)]. Maximum release is determined by lysis of targets by detergent (2% Triton X-100; Sigma Chemical Co., St. Louis, Mo.). Spontaneous release is <25% of maximum release for all experiments.
The results of such an analysis indicate the extent to which HLA-restricted CTL populations have been stimulated by previous exposure to 184P1E2 or a 184P1E2 vaccine.
Similarly, Class II restricted HTL responses may also be analyzed. Purified PBMC are cultured in a 96-well flat bottom plate at a density of 1.5×105 cells/well and are stimulated with 10 μg/ml synthetic peptide of the invention, whole 184P1E2 antigen, or PHA. Cells are routinely plated in replicates of 4-6 wells for each condition. After seven days of culture, the medium is removed and replaced with fresh medium containing 10 U/ml IL-2. Two days later, 1 μCi 3H-thymidine is added to each well and incubation is continued for an additional 18 hours. Cellular DNA is then harvested on glass fiber mats and analyzed for 3H-thymidine incorporation. Antigen-specific T cell proliferation is calculated as the ratio of 3H-thymidine incorporation in the presence of antigen divided by the 3H-thymidine incorporation in the absence of antigen.
Example 29
Induction of Specific CTL Response in Humans
A human clinical trial for an immunogenic composition comprising CTL and HTL epitopes of the invention is set up as an IND Phase I, dose escalation study and carried out as a randomized, double-blind, placebo-controlled trial. Such a trial is designed, for example, as follows:
A total of about 27 individuals are enrolled and divided into 3 groups:
Group I: 3 subjects are injected with placebo and 6 subjects are injected with 5 μg of peptide composition;
Group II: 3 subjects are injected with placebo and 6 subjects are injected with 50 μg peptide composition;
Group III: 3 subjects are injected with placebo and 6 subjects are injected with 500 μg of peptide composition.
After 4 weeks following the first injection, all subjects receive a booster inoculation at the same dosage.
The endpoints measured in this study relate to the safety and tolerability of the peptide composition as well as its immunogenicity. Cellular immune responses to the peptide composition are an index of the intrinsic activity of this the peptide composition, and can therefore be viewed as a measure of biological efficacy. The following summarize the clinical and laboratory data that relate to safety and efficacy endpoints.
Safety: The incidence of adverse events is monitored in the placebo and drug treatment group and assessed in terms of degree and reversibility.
Evaluation of Vaccine Efficacy: For evaluation of vaccine efficacy, subjects are bled before and after injection. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.
The vaccine is found to be both safe and efficacious.
Example 30
Phase II Trials in Patients Expressing 184P1E2
Phase II trials are performed to study the effect of administering the CTL-HTL peptide compositions to patients having cancer that expresses 184P1E2. The main objectives of the trial are to determine an effective dose and regimen for inducing CTLs in cancer patients that express 184P1E2, to establish the safety of inducing a CTL and HTL response in these patients, and to see to what extent activation of CTLs improves the clinical picture of these patients, as manifested, e.g., by the reduction and/or shrinking of lesions. Such a study is designed, for example, as follows:
The studies are performed in multiple centers. The trial design is an open-label, uncontrolled, dose escalation protocol wherein the peptide composition is administered as a single dose followed six weeks later by a single booster shot of the same dose. The dosages are 50, 500 and 5,000 micrograms per injection. Drug-associated adverse effects (severity and reversibility) are recorded.
There are three patient groupings. The first group is injected with 50 micrograms of the peptide composition and the second and third groups with 500 and 5,000 micrograms of peptide composition, respectively. The patients within each group range in age from 21-65 and represent diverse ethnic backgrounds. All of them have a tumor that expresses 184P1E2.
Clinical manifestations or antigen-specific T-cell responses are monitored to assess the effects of administering the peptide compositions. The vaccine composition is found to be both safe and efficacious in the treatment of 184P1E2-associated disease.
Example 31
Induction of CTL Responses Using a Prime Boost Protocol
A prime boost protocol similar in its underlying principle to that used to confirm the efficacy of a DNA vaccine in transgenic mice, such as described above in the Example entitled “The Plasmid Construct and the Degree to Which It Induces Immunogenicity,” can also be used for the administration of the vaccine to humans. Such a vaccine regimen can include an initial administration of, for example, naked DNA followed by a boost using recombinant virus encoding the vaccine, or recombinant protein/polypeptide or a peptide mixture administered in an adjuvant.
For example, the initial immunization may be performed using an expression vector, such as that constructed in the Example entitled “Construction of “Minigene” Multi-Epitope DNA Plasmids” in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose is then administered. The booster can be recombinant fowlpox virus administered at a dose of 5-107 to 5×109 pfu. An alternative recombinant virus, such as an MVA, canarypox, adenovirus, or adeno-associated virus, can also be used for the booster, or the polyepitopic protein or a mixture of the peptides can be administered. For evaluation of vaccine efficacy, patient blood samples are obtained before immunization as well as at intervals following administration of the initial vaccine and booster doses of the vaccine. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.
Analysis of the results indicates that a magnitude of response sufficient to achieve a therapeutic or protective immunity against 184P1E2 is generated.
Example 32
Administration of Vaccine Compositions Using Dendritic Cells (DC)
Vaccines comprising peptide epitopes of the invention can be administered using APCs, or “professional” APCs such as DC. In this example, peptide-pulsed DC are administered to a patient to stimulate a CTL response in vivo. In this method, dendritic cells are isolated, expanded, and pulsed with a vaccine comprising peptide CTL and HTL epitopes of the invention. The dendritic cells are infused back into the patient to elicit CTL and HTL responses in vivo. The induced CTL and HTL then destroy or facilitate destruction, respectively, of the target cells that bear the 184P1E2 protein from which the epitopes in the vaccine are derived.
For example, a cocktail of epitope-comprising peptides is administered ex vivo to PBMC, or isolated DC therefrom. A pharmaceutical to facilitate harvesting of DC can be used, such as ProgenipoietinTM (Monsanto, St. Louis, Mo.) or GM-CSF/IL-4. After pulsing the DC with peptides, and prior to reinfusion into patients, the DC are washed to remove unbound peptides.
As appreciated clinically, and readily determined by one of skill based on clinical outcomes, the number of DC reinfused into the patient can vary (see, e.g., Nature Med. 4:328, 1998; Nature Med. 2:52, 1996 and Prostate 32:272, 1997). Although 2-50×106 DC per patient are typically administered, larger number of DC, such as 107 or 108 can also be provided. Such cell populations typically contain between 50-90% DC.
In some embodiments, peptide-loaded PBMC are injected into patients without purification of the DC. For example, PBMC generated after treatment with an agent such as ProgenipoietinTM are injected into patients without purification of the DC. The total number of PBMC that are administered often ranges from 108 to 1010. Generally, the cell doses injected into patients is based on the percentage of DC in the blood of each patient, as determined, for example, by immunofluorescence analysis with specific anti-DC antibodies. Thus, for example, if ProgenipoietinTM mobilizes 2% DC in the peripheral blood of a given patient, and that patient is to receive 5×106 DC, then the patient will be injected with a total of 2.5×108 peptide-loaded PBMC. The percent DC mobilized by an agent such as ProgenipoietinTM is typically estimated to be between 2-10%, but can vary as appreciated by one of skill in the art.
Ex Vivo Activation of CTL/HTL Responses
Alternatively, ex vivo CTL or HTL responses to 184P1E2 antigens can be induced by incubating, in tissue culture, the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of APC, such as DC, and immunogenic peptides. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cells, i.e., tumor cells.
Example 33
An Alternative Method of Identifying and Confirming Motif-Bearing Peptides
Another method of identifying and confirming motif-bearing peptides is to elute them from cells bearing defined MHC molecules. For example, EBV transformed B cell lines used for tissue typing have been extensively characterized to determine which HLA molecules they express. In certain cases these cells express only a single type of HLA molecule. These cells can be transfected with nucleic acids that express the antigen of interest, e.g. 184P1E2. Peptides produced by endogenous antigen processing of peptides produced as a result of transfection will then bind to HLA molecules within the cell and be transported and displayed on the cell's surface. Peptides are then eluted from the HLA molecules by exposure to mild acid conditions and their amino acid sequence determined, e.g., by mass spectral analysis (e.g., Kubo et al., J. Immunol. 152:3913, 1994). Because the majority of peptides that bind a particular HLA molecule are motif-bearing, this is an alternative modality for obtaining the motif-bearing peptides correlated with the particular HLA molecule expressed on the cell.
Alternatively, cell lines that do not express endogenous HLA molecules can be transfected with an expression construct encoding a single HLA allele. These cells can then be used as described, i.e., they can then be transfected with nucleic acids that encode 184P1E2 to isolate peptides corresponding to 184P1E2 that have been presented on the cell surface. Peptides obtained from such an analysis will bear motif(s) that correspond to binding to the single HLA allele that is expressed in the cell.
As appreciated by one in the art, one can perform a similar analysis on a cell bearing more than one
HLA allele and subsequently determine peptides specific for each HLA allele expressed. Moreover, one of skill would also recognize that means other than transfection, such as loading with a protein antigen, can be used to provide a source of antigen to the cell.
Example 34
Complementary Polynucleotides
Sequences complementary to the 184P1E2-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring 184P1E2. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using, e.g., OLIGO 4.06 software (National Biosciences) and the coding sequence of 184P1E2. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5′ sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to a 184P1E2-encoding transcript.
Example 35
Purification of Naturally-Occurring or Recombinant 184P1E2 Using 184P1E2-Specific Antibodies
Naturally occurring or recombinant 184P1E2 is substantially purified by immunoaffinity chromatography using antibodies specific for 184P1E2. An immunoaffinity column is constructed by covalently coupling anti-184P1E2 antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.
Media containing 184P1E2 are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of 184P1E2 (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/184P1E2 binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and GCR.P is collected.
Example 36
Identification of Molecules which Interact with 184P1E2
184P1E2, or biologically active fragments thereof, are labeled with 121 1 Bolton-Hunter reagent. (See, e.g., Bolton et al. (1973) Biochem. J. 133:529.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled 184P1E2, washed, and any wells with labeled 184P1E2 complex are assayed. Data obtained using different concentrations of 184P1E2 are used to calculate values for the number, affinity, and association of 184P1E2 with the candidate molecules.
Example 37
In Vivo Assay for 184P1E2 Tumor Growth Promotion
The effect of the 184P1E2 protein on tumor cell growth is evaluated in vivo by evaluating tumor development and growth of cells expressing or lacking 184P1E2. For example, SCID mice are injected subcutaneously on each flank with 1×106 of either 3T3, bladder or lung cancer cell lines (e.g. UM-UC3, J82, CaLu1 and A427 cells) containing tkNeo empty vector or 184P1E2. At least two strategies may be used: (1) Constitutive 184P1E2 expression under regulation of a promoter such as a constitutive promoter obtained from the genomes of viruses such as polyoma virus, fowipox virus (UK 2,211,504 published 5 Jul. 1989), adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40 (SV40), or from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, provided such promoters are compatible with the host cell systems, and (2) Regulated expression under control of an inducible vector system, such as ecdysone, tetracycline, etc., provided such promoters are compatible with the host cell systems. Tumor volume is then monitored by caliper measurement at the appearance of palpable tumors and followed over time to determine if 184P1E2-expressing cells grow at a faster rate and whether tumors produced by 184P1E2-expressing cells demonstrate characteristics of altered aggressiveness (e.g. enhanced metastasis, vascularization, reduced responsiveness to chemotherapeutic drugs).
Additionally, mice can be implanted with 1×105 of the same cells orthotopically to determine if 184P1E2 has an effect on local growth in the bladder or lung, and whether 184P1E2 affects the ability of the cells to metastasize, specifically to lymph nodes, adrenal, liver and bone (Miki T et al, Oncol Res. 2001; 12:209; Fu X et al, Int J Cancer. 1991, 49:938).
The assay is also useful to determine the 184P1E2 inhibitory effect of candidate therapeutic compositions, such as for example, 184P1E2 intrabodies, 184P1E2 antisense molecules and ribozymes.
Example 38
184P1E2 Monoclonal Antibody-mediated Inhibition of Bladder and Lung Tumors In Vivo
The significant expression of 184P1E2 in cancer tissues, together with its restrictive expression in normal tissues makes 184P1E2 a good target for antibody therapy. Similarly, 184P1E2 is a target for T cell-based immunotherapy. Thus, the therapeutic efficacy of anti-184P1E2 mAbs in human bladder cancer xenograft mouse models is evaluated by using recombinant cell lines such as UM-UC3-184P1E2, J82-184P1E2, and 3T3-184P1E2 (see, e.g., Kaighn, M. E., et al., Invest Urol, 1979. 17(1): p. 16-23). Similarly, anti-184P1E2 mAbs are evaluated in human lung cancer xenograft models using recombinant cell lines such as CaLU-184P1E2 and A427-184P1E2.
Antibody efficacy on tumor growth and metastasis formation is studied, e.g., in a mouse orthotopic bladder cancer xenograft models and mouse lung xenograft models. The antibodies can be unconjugated, as discussed in this Example, or can be conjugated to a therapeutic modality, as appreciated in the art. Anti-184P1E2 mAbs inhibit formation of both lung and bladder xenografts. Anti-184P1E2 mAbs also retard the growth of established orthotopic tumors and prolonged survival of tumor-bearing mice. These results indicate the utility of anti-184P1E2 mAbs in the treatment of local and advanced stages of lung and bladder cancer. (See, e.g., Saffran, D., et al., PNAS 10: 1073-1078).
Administration of the anti-184P1E2 mAbs led to retardation of established orthotopic tumor growth and inhibition of metastasis to distant sites, resulting in a significant prolongation in the survival of tumor-bearing mice. These studies indicate that 184P1E2 as an attractive target for immunotherapy and demonstrate the therapeutic potential of anti-184P1E2 mAbs for the treatment of local and metastatic prostate cancer. This example demonstrates that unconjugated 184P1E2 monoclonal antibodies are effective to inhibit the growth of human bladder tumor xenografts and human lung xenografts grown in SCID mice; accordingly a combination of such efficacious monoclonal antibodies is also effective.
Tumor Inhibition Using Multiple Unconjugated 184P1E2 mAbs
Materials and Methods
184P1E2 Monoclonal Antibodies:
Monoclonal antibodies are raised against 184P1E2 as described in the Example entitled “Generation of 184P1E2 Monoclonal Antibodies (mAbs).” The antibodies are characterized by ELISA, Western blot, FACS, and immunoprecipitation for their capacity to bind 184P1E2. Epitope mapping data for the anti-184P1E2 mAbs, as determined by ELISA and Western analysis, recognize epitopes on the 184P1E2 protein. Immunohistochemical analysis of prostate cancer tissues and cells with these antibodies is performed.
The monoclonal antibodies are purified from ascites or hybridoma tissue culture supernatants by Protein-G Sepharose chromatography, dialyzed against PBS, filter sterilized, and stored at −20° C. Protein determinations are performed by a Bradford assay (Bio-Rad, Hercules, Calif.). A therapeutic monoclonal antibody or a cocktail comprising a mixture of individual monoclonal antibodies is prepared and used for the treatment of mice receiving subcutaneous or orthotopic injections of UM-UC3 and CaLu1 tumor xenografts.
Cell Lines
The balder and lung carcinoma cell lines, UM-UC3, J82, CaLu1 and A427 as well as the fibroblast line NIH 3T3 (American Type Culture Collection) are maintained in DMEM supplemented with L-glutamine and 10% FBS.
A UM-UC3-184P1E2, J82-184P1E2, CaLu1-184P1E2, A427-184P1E2 and 3T3-184P1E2 cell populations are generated by retroviral gene transfer as described in Hubert, R. S., et al., Proc Natl Acad Sci USA, 1999. 96(25): 14523.
Xenograft Mouse Models.
Subcutaneous (s.c.) tumors are generated by injection of 1×106 cancer cells mixed at a 1:1 dilution with Matrigel (Collaborative Research) in the right flank of male SCID mice. To test antibody efficacy on tumor formation, i.p. antibody injections are started on the same day as tumor-cell injections. As a control, mice are injected with either purified mouse IgG (ICN) or PBS; or a purified monoclonal antibody that recognizes an irrelevant antigen not expressed in human cells. In preliminary studies, no difference is found between mouse IgG or PBS on tumor growth. Tumor sizes are determined by caliper measurements, and the tumor volume is calculated as length×width×height. Mice with s.c. tumors greater than 1.5 cm in diameter are sacrificed.
Orthotopic injections are performed under anesthesia by using ketamine/xylazine. For bladder orthotopic studies, an incision is made through the abdomen to expose the bladder and tumor cells (5×105) mixed with Matrigel are injected into the bladder capsule in a 10-μl volume. To monitor tumor growth, mice are palpated and blood is collected on a weekly basis to measure BTA levels. For lung orthopotic models, an incision is made through the abdominal muscles to expose the lung. Tumor cells (5×105) mixed with matrigel are injected into the bronchioalveolar region of the right lung (McLemore T L et al, Cancer Res. 1988; 48:2880). To monitor tumor growth, blood is collected on a weekly basis to measure CA 125 levels. The mice are segregated into groups for the appropriate treatments, with anti-184P1E2 or control mAbs being injected i.p.
Anti-184P1E2 mAbs Inhibit Growth of 184P1E2-Expressing Xenograft-Cancer Tumors
The effect of anti-184P1E2 mAbs on tumor formation is tested by using UM-UC3 and CaLu1 orthotopic models. As compared with the s.c. tumor model, the orthotopic model, which requires injection of tumor cells directly in the mouse bladder and lung, respectively, results in a local tumor growth, development of metastasis in distal sites, deterioration of mouse health, and subsequent death (Saffran, D., et al., PNAS supra; Fu, X., et al., Int J Cancer, 1992. 52(6): p. 987-90; Kubota, T., J Cell Biochem, 1994. 56(1): p. 4-8). The features make the orthotopic model more representative of human disease progression and allowed us to follow the therapeutic effect of mAbs on clinically relevant end points.
Accordingly, tumor cells are injected into the mouse bladder or lung, and 2 days later, the mice are segregated into two groups and treated with either: a) 200-500 μg, of anti-184P1E2 Ab, or b) PBS three times per week for two to five weeks.
A major advantage of the orthotopic cancer models is the ability to study the development of metastases. Formation of metastasis in mice bearing established orthotopic tumors is studies by IHC analysis on lung sections using an antibody against a tumor-specific cell-surface protein such as anti-CK20 for bladder cancer and anti-CEA antibody for lung cancer models (Lin S et al, Cancer Detect Prev. 2001; 25:202).
Mice bearing established orthotopic tumors are administered 1000 μg injections of either anti-184P1E2 mAb or PBS over a 4-week period. Mice in both groups are allowed to establish a high tumor burden, to ensure a high frequency of metastasis formation in mouse lungs. Mice then are killed and their bladders, livers, bone and lungs are analyzed for the presence of tumor cells by IHC analysis.
These studies demonstrate a broad anti-tumor efficacy of anti-184P1E2 antibodies on initiation and progression of prostate and kidney cancer in xenograft mouse models. Anti-184P1E2 antibodies inhibit tumor formation of tumors as well as retarding the growth of already established tumors and prolong the survival of treated mice. Moreover, anti-184P1E2 mAbs demonstrate a dramatic inhibitory effect on the spread of local bladder and lung tumor to distal sites, even in the presence of a large tumor burden. Thus, anti-184P1E2 mAbs are efficacious on major clinically relevant end points (tumor growth), prolongation of survival, and health.
Example 39
Therapeutic and Diagnostic Use of Anti-184P1E2 Antibodies in Humans
Anti-184P1E2 monoclonal antibodies are safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic purposes in humans. Western blot and immunohistochemical analysis of cancer tissues and cancer xenografts with anti-184P1E2 mAb show strong extensive staining in carcinoma but significantly lower or undetectable levels in normal tissues. Detection of 184P1E2 in carcinoma and in metastatic disease demonstrates the usefulness of the mAb as a diagnostic and/or prognostic indicator. Anti-184P1E2 antibodies are therefore used in diagnostic applications such as immunohistochemistry of kidney biopsy specimens to detect cancer from suspect patients.
As determined by flow cytometry, anti-184P1E2 mAb specifically binds to carcinoma cells. Thus, anti-184P1E2 antibodies are used in diagnostic whole body imaging applications, such as radioimmunoscintigraphy and radioimmunotherapy, (see, e.g., Potamianos S., et. al. Anticancer Res 20(2A):925-948 (2000)) for the detection of localized and metastatic cancers that exhibit expression of 184P1E2. Shedding or release of an extracellular domain of 184P1E2 into the extracellular milieu, such as that seen for alkaline phosphodiesterase B10 (Meerson, N. R., Hepatology 27:563-568 (1998)), allows diagnostic detection of 184P1E2 by anti-184P1E2 antibodies in serum and/or urine samples from suspect patients.
Anti-184P1E2 antibodies that specifically bind 184P1E2 are used in therapeutic applications for the treatment of cancers that express 184P1E2. Anti-184P1E2 antibodies are used as an unconjugated modality and as conjugated form in which the antibodies are attached to one of various therapeutic or imaging modalities well known in the art, such as a prodrugs, enzymes or radioisotopes. In preclinical studies, unconjugated and conjugated anti-184P1E2 antibodies are tested for efficacy of tumor prevention and growth inhibition in the SCID mouse cancer xenograft models, e.g., kidney cancer models AGS-K3 and AGS-K6, (see, e.g., the Example entitled “184P1E2 Monoclonal Antibody-mediated Inhibition of Bladder and Lung Tumors In Vivo”). Conjugated and unconjugated anti-184P1E2 antibodies are used as a therapeutic modality in human clinical trials either alone or in combination with other treatments as described in following Examples.
Example 40
Human Clinical Trials for the Treatment and Diagnosis of Human Carcinomas through Use of Human Anti-184P1E2 Antibodies In Vivo
Antibodies are used in accordance with the present invention which recognize an epitope on 184P1E2, and are used in the treatment of certain tumors such as those listed in Table I. Based upon a number of factors, including 184P1E2 expression levels, tumors such as those listed in Table I are presently preferred indications. In connection with each of these indications, three clinical approaches are successfully pursued.
I.) Adjunctive Therapy:
In adjunctive therapy, patients are treated with anti-184P1E2 antibodies in combination with a chemotherapeutic or antineoplastic agent and/or radiation therapy. Primary cancer targets, such as those listed in Table I, are treated under standard protocols by the addition anti-184P1E2 antibodies to standard first and second line therapy. Protocol designs address effectiveness as assessed by reduction in tumor mass as well as the ability to reduce usual doses of standard chemotherapy. These dosage reductions allow additional and/or prolonged therapy by reducing dose-related toxicity of the chemotherapeutic agent. Anti-184P1E2 antibodies are utilized in several adjunctive clinical trials in combination with the chemotherapeutic or antineoplastic agents adriamycin (advanced prostrate carcinoma), cisplatin (advanced head and neck and lung carcinomas), taxol (breast cancer), and doxorubicin (preclinical).
II.) Monotherapy:
In connection with the use of the anti-184P1E2 antibodies in monotherapy of tumors, the antibodies are administered to patients without a chemotherapeutic or antineoplastic agent. In one embodiment, monotherapy is conducted clinically in end stage cancer patients with extensive metastatic disease. Patients show some disease stabilization. Trials demonstrate an effect in refractory patients with cancerous tumors.
III.) Imaging Agent:
Through binding a radionuclide (e.g., iodine or yttrium (I131, Y90) to anti-184P1E2 antibodies, the radiolabeled antibodies are utilized as a diagnostic and/or imaging agent. In such a role, the labeled antibodies localize to both solid tumors, as well as, metastatic lesions of cells expressing 184P1E2. In connection with the use of the anti-184P1E2 antibodies as imaging agents, the antibodies are used as an adjunct to surgical treatment of solid tumors, as both a pre-surgical screen as well as a post-operative follow-up to determine what tumor remains and/or returns. In one embodiment, a (111In)-184P1E2 antibody is used as an imaging agent in a Phase I human clinical trial in patients having a carcinoma that expresses 184P1E2 (by analogy see, e.g., Divgi et al. J. Natl. Cancer Inst. 83:97-104 (1991)). Patients are followed with standard anterior and posterior gamma camera. The results indicate that primary lesions and metastatic lesions are identified
Dose and Route of Administration
As appreciated by those of ordinary skill in the art, dosing considerations can be determined through comparison with the analogous products that are in the clinic. Thus, anti-184P1E2 antibodies can be administered with doses in the range of 5 to 400 mg/m2, with the lower doses used, e.g., in connection with safety studies. The affinity of anti-184P1E2 antibodies relative to the affinity of a known antibody for its target is one parameter used by those of skill in the art for determining analogous dose regimens. Further, anti-184P1E2 antibodies that are fully human antibodies, as compared to the chimeric antibody, have slower clearance; accordingly, dosing in patients with such fully human anti-184P1E2 antibodies can be lower, perhaps in the range of 50 to 300 mg/m2, and still remain efficacious. Dosing in mg/m2, as opposed to the conventional measurement of dose in mg/kg, is a measurement based on surface area and is a convenient dosing measurement that is designed to include patients of all sizes from infants to adults.
Three distinct delivery approaches are useful for delivery of anti-184P1E2 antibodies. Conventional intravenous delivery is one standard delivery technique for many tumors. However, in connection with tumors in the peritoneal cavity, such as tumors of the ovaries, biliary duct, other ducts, and the like, intraperitoneal administration may prove favorable for obtaining high dose of antibody at the tumor and to also minimize antibody clearance. In a similar manner, certain solid tumors possess vasculature that is appropriate for regional perfusion. Regional perfusion allows for a high dose of antibody at the site of a tumor and minimizes short term clearance of the antibody.
Clinical Development Plan (CDP)
Overview: The CDP follows and develops treatments of anti-184P1E2 antibodies in connection with adjunctive therapy, monotherapy, and as an imaging agent. Trials initially demonstrate safety and thereafter confirm efficacy in repeat doses. Trails are open label comparing standard chemotherapy with standard therapy plus anti-184P1E2 antibodies. As will be appreciated, one criteria that can be utilized in connection with enrollment of patients is 184P1E2 expression levels in their tumors as determined by biopsy.
As with any protein or antibody infusion-based therapeutic, safety concerns are related primarily to (i) cytokine release syndrome, i.e., hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 184P1E2. Standard tests and follow-up are utilized to monitor each of these safety concerns. Anti-184P1E2 antibodies are found to be safe upon human administration.
Example 41
Human Clinical Trial Adjunctive Therapy with Human Anti-184P1E2 Antibody and Chemotherapeutic Agent
A phase I human clinical trial is initiated to assess the safety of six intravenous doses of a human anti-184P1E2 antibody in connection with the treatment of a solid tumor, e.g., a cancer of a tissue listed in Table I. In the study, the safety of single doses of anti-184P1E2 antibodies when utilized as an adjunctive therapy to an antineoplastic or chemotherapeutic agent, such as cisplatin, topotecan, doxorubicin, adriamycin, taxol, or the like, is assessed. The trial design includes delivery of six single doses of an anti-184P1E2 antibody with dosage of antibody escalating from approximately about 25 mg/m2 to about 275 mg/m2 over the course of the treatment in accordance with the following schedule:


















Day 0
Day 7
Day 14
Day 21
Day 28
Day 35






















mAb Dose
25
75
125
175
225
275



mg/m2
mg/m2
mg/m2
mg/m2
mg/m2
mg/m2


Chemotherapy
+
+
+
+
+
+


(standard dose)









Patients are closely followed for one-week following each administration of antibody and chemotherapy. In particular, patients are assessed for the safety concerns mentioned above: (i) cytokine release syndrome, i.e., hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the human antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 184P1E2. Standard tests and follow-up are utilized to monitor each of these safety concerns. Patients are also assessed for clinical outcome, and particularly reduction in tumor mass as evidenced by MRI or other imaging.
The anti-184P1E2 antibodies are demonstrated to be safe and efficacious, Phase II trials confirm the efficacy and refine optimum dosing.
Example 42
Human Clinical Trial: Monotherapy with Human Anti-184P1E2 Antibody
Anti-184P1E2 antibodies are safe in connection with the above-discussed adjunctive trial, a Phase II human clinical trial confirms the efficacy and optimum dosing for monotherapy. Such trial is accomplished, and entails the same safety and outcome analyses, to the above-described adjunctive trial with the exception being that patients do not receive chemotherapy concurrently with the receipt of doses of anti-184P1E2 antibodies.
Example 43
Human Clinical Trial: Diagnostic Imaging with Anti-184P1E2 Antibody
Once again, as the adjunctive therapy discussed above is safe within the safety criteria discussed above, a human clinical trial is conducted concerning the use of anti-184P1E2 antibodies as a diagnostic imaging agent. The protocol is designed in a substantially similar manner to those described in the art, such as in Divgi et al. J. Natl. Cancer Inst. 83:97-104 (1991). The antibodies are found to be both safe and efficacious when used as a diagnostic modality.
Example 44
Homology Comparison of 184P1E2 to Known Sequences
The 184P1E2 gene is homologous to a previously cloned gene, namely human peptidylarginine deiminase type III (gi 7706447). The 184P1E2 v.1 and 184P1E2 v.2 shows 99% identity to the published peptidylarginine deiminase type III over the length of the protein (FIG. 4B and Table LIII). While 184P1E2 v.1 differs from gi 7706447 by one amino acid at position 480 (FIG. 4B), 184P1E2 v.2 differs from gi 7706447 by two amino acids at positions 304 and 480 (Table LIII). In contrast, 184P1E2 v.3 is 100% identical to the published peptidylarginine deiminase type III (See FIG. 4B and Table LIII). This indicates that 184P1E2 v.1, v.2 and v.3 represent SNPs of the same gene (see Table LIII). The homology to peptidylarginine deiminase is maintained across species, as 184P1E2 is strongly homologous to mouse and rat peptidylarginine deiminase type III (FIGS. 4C and 4D). The 184P1E2 protein consists of 664 amino acids, with calculated molecular weight of 74.7 kDa, and pI of 5.3. 184P1E2 is an intracellular protein, with localization to the mitochondria and cytosol. 184P1E2 can also localize to the nucleus. Motif analysis revealed the presence of a protein arginine deaminase motif (PAD) over the entire length of the protein, and a cadherin signature at amino acid 314-362 (Table XXI).
Protein arginine deaminases represent a family of 4 arginine deaminase isoforms, all of which catalyses the post-translational conversion of arginine to citrulline residues in a calcium dependent manner (Kanno T et al, J Inv. Dermatol 2000, 115:813). Peptidylarginine deiminase III is also known as the hai follicle type of PAD as it is primarily expressed in epidermis and hair follicles (Watanabe K et al. Biochim Biophys Acta 1988, 966:375). The conversion of arginine to citrulline by PAD has profound effects on the primary structure of target proteins and their biological function. For example, deimination of myelin basic protein enhances its susceptibility to degradation by cathepsin, a condition associated with the pathology of multiple sclerosis (Pritzker L et al, Biochemistry 2000, 39:5382). Several epidermal proteins are deiminated by peptidylarginine deiminase III, including filaggrin, trichohyalin and keratin (Senshu T et al, Biochem. Biophy. Res. Comm 1996, 225:712). Deimination of these substrates leads to their denaturation and to the eventual loss of cell integrity (Mizoguchi M et al, J. Histochem. Cytockem 1998, 46:1303). Peptidylarginine deaminase also acts as a regulator of cell proliferation and survival in some but not all tumor cells. While some tumor lines, such a acute leukemias, respond to peptidylarginine deaminase by undergoing cell arrest at G1 and/or S phases of the cell cycle and apoptosis, other cells are not affected by peptidylarginine deaminase (Gong H et al, Leukemia 2000, 14:826). In other cases, peptidylarginine deaminase provides a protective effect against apoptosis, such as in prostate cancer cells treated with taxol (Kang S et al, Mol Cell 2000, 10:331).
As mentioned above, a cadherin motif was identified at aa 316-342 of the 184P1E2 protein. Cadherins are a family of proteins that function in calcium-dependent cell adhesion. Cadherins preferentially interact with themselves, regulating cell adhesion and tight junctions (Nagafuchi A. Curr Opin Cell Biol. 2001, 13:600). Disruption of cadherin function results in unregulated cell growth and migration, often observed in cancer (Thiery J P, Chopin D. Cancer Metastasis Rev. 1999; 18:31; ). As 184P1E2 is an intracellular protein, it is unlikely to, by itself, mediate cell-cell adhesion. However, the presence of a cadherin motif suggests that 184P1E2 participate in protein-protein interactions.
The presence of a peptidylarginine deaminase motif and protein-protein interaction domain along with its localization indicate that 184P1E2 functions in regulating protein interactions and signal transduction in mammalian cells, thereby regulating cell proliferation, survival, differentiation as well as, gene expression. These biological functions have a direct effect on tumor growth and progression.
Accordingly, when 184P1E2 functions as a regulator of cell growth, tumor formation, cell signaling or as a modulator of transcription involved in activating genes associated with survival, invasion, tumorigenesis or proliferation, 184P1E2 is used for therapeutic, diagnostic, prognostic and/or preventative purposes. In addition, when a molecule, such as a variant or SNP of 184P1E2 is expressed in cancerous tissues, such as those listed in Table I, they are used for therapeutic, diagnostic, prognostic and/or preventative purposes.
Example 45
Regulation of Transcription
The localization of 184P1E2 coupled to the presence of protein interaction domains within its sequence indicate that 184P1E2 modulates the transcriptional regulation of eukaryotic genes. Regulation of gene expression is confirmed, e.g., by studying gene expression in cells expressing or lacking 184P1E2. For this purpose, two types of experiments are performed.
In the first set of experiments, RNA from parental and 184P1E2-expressing cells are extracted and hybridized to commercially available gene arrays (Clontech) (Smid-Koopman E et al. Br J Cancer. 2000. 83:246). Resting cells as well as cells treated with FBS, androgen or growth factors are compared. Differentially expressed genes are identified in accordance with procedures known in the art. The differentially expressed genes are then mapped to biological pathways (Chen K et al. Thyroid. 2001. 11:41.).
In the second set of experiments, specific transcriptional pathway activation is evaluated using commercially available (Stratagene) luciferase reporter constructs including: NFkB-luc, SRE-luc, ELK1-luc, ARE-luc, p53-luc, and CRE-luc. These transcriptional reporters contain consensus binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways, and represent a good tool to ascertain pathway activation and screen for positive and negative modulators of pathway activation.
Thus, 184P1E2 plays a role in gene regulation, and it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.
Example 46
Identification and Confirmation of Potential Signal Transduction Pathways
Many mammalian proteins have been reported to interact with signaling molecules and to participate in regulating signaling pathways. (J Neurochem. 2001; 76:217-223). Cadherins in particular have been associated with the β-catenin signaling cascade that control cell transformation and invasion (Gottardi C J et al, J Cell Biol. 2001, 153:1049). Based on the presence of a cadherin motif 184P1E2 regulates signaling pathways important for cell growth and invasion. In addition, the 184P1E2 protein contains several phosphorylation sites (see Table XX) indicating an association with specific signaling cascades. Using immunoprecipitation and Western blotting techniques, proteins are identified that associate with 184P1E2 and mediate signaling events. Several pathways known to play a role in cancer biology can be regulated by 184P1E2, including phospholipid pathways such as PI3K, AKT, etc, adhesion and migration pathways, including FAK, Rho, Rac-1, β-catenin, etc, as well as mitogenic/survival cascades such as ERK, p38, etc (Cell Growth Differ. 2000, 11:279; J Biol Chem. 1999, 274:801; Oncogene. 2000, 19:3003, J. Cell Biol. 1997, 138:913.).
To confirm that 184P1E2 directly or indirectly activates known signal transduction pathways in cells, luciferase (luc) based transcriptional reporter assays are carried out in cells expressing individual genes. These transcriptional reporters contain consensus-binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways. The reporters and examples of these associated transcription factors, signal transduction pathways, and activation stimuli are listed below.

    
    
        1. NFkB-luc, NFkB/Rel; Ik-kinase/SAPK; growth/apoptosis/stress
        2. SRE-luc, SRF/TCF/ELK1; MAPK/SAPK; growth/differentiation
        3. AP-1-luc, FOS/JUN; MAPK/SAPK/PKC; growth/apoptosis/stress
        4. RE-luc, androgen receptor; steroids/MAPK; growth/differentiation/apoptosis
        5. p53-luc, p53; SAPK; growth/differentiation/apoptosis
        6. CRE-luc, CREB/ATF2; PKA/p38; growth/apoptosis/stress
        7. TCF-luc, TCF/Lef; β-catenin, Adhesion/invasion
    
    


Gene-mediated effects can be assayed in cells showing mRNA expression. Luciferase reporter plasmids can be introduced by lipid-mediated transfection (TFX-50, Promega). Luciferase activity, an indicator of relative transcriptional activity, is measured by incubation of cell extracts with luciferin substrate and luminescence of the reaction is monitored in a luminometer.
Signaling pathways activated by 184P1E2 are mapped and used for the identification and validation of therapeutic targets. When 184P1E2 is involved in cell signaling, it is used as target for diagnostic, prognostic, preventative and/or therapeutic purposes.
Example 47
Involvement in Tumor Progression
Based on the documented role of peptidylarginine deiminase in cell growth, proliferation and survival (Kang S et al, Mol Cell 2000, 10:331), the 184P1E2 gene can contribute to the growth of cancer cells. The role of 184P1E2 in tumor growth is confirmed in a variety of primary and transfected cell lines including bladder and lung cell lines, as well as NIH 3T3 cells engineered to stably express 184P1E2. Parental cells lacking 184P1E2 and cells expressing 184P1E2 are evaluated for cell growth using a well-documented proliferation assay (Fraser S P, Grimes J A, Djamgoz M B. Prostate. 2000; 44:61, Johnson D E, Ochieng J, Evans S L. Anticancer Drugs. 1996, 7:288).
To confirm the role of 184P1E2 in the transformation process, its effect in colony forming assays is investigated. Parental NIH-3T3 cells lacking 184P1E2 are compared to NIH-3T3 cells expressing 184P1E2, using a soft agar assay under stringent and more permissive conditions (Song Z. et al. Cancer Res. 2000; 60:6730).
To confirm the role of 184P1E2 in invasion and metastasis of cancer cells, a well-established assay is used, e.g., a Transwell Insert System assay (Becton Dickinson) (Cancer Res. 1999; 59:6010). Control cells, including prostate, breast and kidney cell lines lacking 184P1E2 are compared to cells expressing 184P1E2. Cells are loaded with the fluorescent dye, calcein, and plated in the top well of the Transwell insert coated with a basement membrane analog. Invasion is determined by fluorescence of cells in the lower chamber relative to the fluorescence of the entire cell population.
184P1E2 can also play a role in cell cycle and apoptosis. Parental cells and cells expressing 184P1E2 are compared for differences in cell cycle regulation using a well-established BrdU assay (Abdel-Malek Z A. J Cell Physiol. 1988, 136:247). In short, cells are grown under both optimal (full serum) and limiting (low serum) conditions are labeled with BrdU and stained with anti-BrdU Ab and propidium iodide. Cells are analyzed for entry into the G1, S, and G2M phases of the cell cycle. Alternatively, the effect of stress on apoptosis is evaluated in control parental cells and cells expressing 184P1E2, including normal and tumor bladder and lung cells. Engineered and parental cells are treated with various chemotherapeutic agents, such as etoposide, taxol, etc, and protein synthesis inhibitors, such as cycloheximide. Cells are stained with annexin V-FITC and cell death is measured by FACS analysis. The modulation of cell death by 184P1E2 can play a critical role in regulating tumor progression and tumor load.
When 184P1E2 plays a role in cell growth, transformation, invasion or apoptosis, it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.
Example 48
Involvement in Angiogenesis
Angiogenesis or new capillary blood vessel formation is necessary for tumor growth (Hanahan D, Folkman J. Cell. 1996, 86:353; Folkman J. Endocrinology. 1998 139:441). Based on the effect of phsophodieseterase inhibitors on endothelial cells, 184P1E2 plays a role in angiogenesis (DeFouw L et al, Microvasc Res 2001, 62:263). Several assays have been developed to measure angiogenesis in vitro and in vivo, such as the tissue culture assays endothelial cell tube formation and endothelial cell proliferation. Using these assays as well as in vitro neo-vascularization, the role of 184P1E2 in angiogenesis, enhancement or inhibition, is confirmed.
For example, endothelial cells engineered to express 184P1E2 are evaluated using tube formation and proliferation assays. The effect of 184P1E2 is also confirmed in animal models in vivo. For example, cells either expressing or lacking 184P1E2 are implanted subcutaneously in immunocompromised mice. Endothelial cell migration and angiogenesis are evaluated 5-15 days later using immunohistochemistry techniques. 184P1E2 affects angiogenesis, and it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes
Example 49
Involvement in Protein-Protein Interactions
Cadherin motifs have been shown to mediate interaction with other proteins, specially similar cadherin proteins, thereby regulating cell adhesion and growth (Cavallaro U et al, Cancer Lett. 2002, 176:123; Kovacs E M et al., Curr Biol. 2002, 12:379). Using immunoprecipitation techniques as well as two yeast hybrid systems, proteins are identified that associate with 184P1E2. Immunoprecipitates from cells expressing 184P1E2 and cells lacking 184P1E2 are compared for specific protein-protein associations.
Studies are performed to confirm the extent of association of 184P1E2 with effector molecules, such as nuclear proteins, transcription factors, kinases, phsophates etc. Studies comparing 184P1E2 positive and 184P1E2 negative cells as well as studies comparing unstimulated/resting cells and cells treated with epithelial cell activators, such as cytokines, growth factors, androgen and anti-integrin Ab reveal unique interactions.
In addition, protein-protein interactions are confirmed using two yeast hybrid methodology (Curr Opin Chem Biol. 1999, 3:64). A vector carrying a library of proteins fused to the activation domain of a transcription factor is introduced into yeast expressing a 184P1E2-DNA-binding domain fusion protein and a reporter construct. Protein-protein interaction is detected by colorimetric reporter activity. Specific association with effector molecules and transcription factors directs one of skill to the mode of action of 184P1E2, and thus identifies therapeutic, prognostic, preventative and/or diagnostic targets for cancer. This and similar assays are also used to identify and screen for small molecules that interact with 184P1E2.
Thus it is found that 184P1E2 associates with proteins and small molecules. Accordingly, 184P1E2 and these proteins and small molecules are used for diagnostic, prognostic, preventative and/or therapeutic purposes.
Example 50
Involvement in Deimination
As previously mentioned, petidylarginine deiminases convert protein-bound arginine to citrulline, thereby altering the structure and function of target proteins (Kanno T et al, J Inv. Dermatol 2000, 115:813). The petidylarginine deiminases of 184P1E2 will be confirmed in recombinant cell lines as well as primary bladder and lung tumor cells. Cells expressing 184P1E2 and control cells lacking 184P1E2 are grown on sterile glass coverslips, and fixed in paraformaldehyde. Citrulline residues located in cellular proteins are chemically altered for better recognition using potassium ferricyanide. Citrulline residues are detected using a citrulline-specific antibody by immunofluorescence.
When 184P1E2 functions as a deiminase, it is used as a target for diagnostic, prognostic, preventative and therapeutic purposes.
Throughout this application, various website data content, publications, patent applications and patents are referenced. (Websites are referenced by their Uniform Resource Locator, or URL, addresses on the World Wide Web.) The disclosures of each of these references are hereby incorporated by reference herein in their entireties.
The present invention is not to be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects of the invention, and any that are functionally equivalent are within the scope of the invention. Various modifications to the models and methods of the invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description and teachings, and are similarly intended to fall within the scope of the invention. Such modifications or other embodiments can be practiced without departing from the true scope and spirit of the invention.







TABLE I





Tissues that Express 184P1E2 When Malignant







Bladder


Kidney


Lung
















TABLE II







Amino Acid Abbreviations











SINGLE LETTER
THREE LETTER
FULL NAME






F
Phe
phenylalanine



L
Leu
leucine



S
Ser
serine



Y
Tyr
tyrosine



C
Cys
cysteine



W
Trp
tryptophan



P
Pro
proline



H
His
histidine



Q
Gln
glutamine



R
Arg
arginine



I
Ile
isoleucine



M
Met
methionine



T
Thr
threonine



N
Asn
asparagine



K
Lys
lysine



V
Val
valine



A
Ala
alanine



D
Asp
aspartic acid



E
Glu
glutamic acid



G
Gly
glycine
















TABLE III





Amino Acid Substitution Matrix


Adapted from the GCG Software 9.0 BLOSUM62 amino acid substitution matrix (block substitution


matrix). The higher the value, the more likely a substitution is found in related, natural proteins.



































A
C
D
E
F
G
H
I
K
L
M
N
P
Q
R
S
T
V
W
Y
.


4
0
−2
−1
−2
0
−2
−1
−1
−1
−1
−2
−1
−1
−1
1
0
0
−3
−2
A



9
−3
−4
−2
−3
−3
−1
−3
−1
−1
−3
−3
−3
−3
−1
−1
−1
−2
−2
C




6
2
−3
−1
−1
−3
−1
−4
−3
1
−1
0
−2
0
−1
−3
−4
−3
D





5
−3
−2
0
−3
1
−3
−2
0
−1
2
0
0
−1
−2
−3
−2
E






6
−3
−1
0
−3
0
0
−3
−4
−3
−3
−2
−2
−1
1
3
F







6
−2
−4
−2
−4
−3
0
−2
−2
−2
0
−2
−3
−2
−3
G








8
−3
−1
−3
−2
1
−2
0
0
−1
−2
−3
−2
2
H









4
−3
2
1
−3
−3
−3
−3
−2
−1
3
−3
−1
I










5
−2
−1
0
−1
1
2
0
−1
−2
−3
−2
K











4
2
−3
−3
−2
−2
−2
−1
1
−2
−1
L












5
−2
−2
0
−1
−1
−1
1
−1
−1
M













6
−2
0
0
1
0
−3
−4
−2
N














7
−1
−2
−1
−1
−2
−4
−3
P















5
1
0
−1
−2
−2
−1
Q
















5
−1
−1
−3
−3
−2
R

















4
1
−2
−3
−2
S


















5
0
−2
−2
T



















4
−3
−1
V




















11
2
W





















7
Y
















TABLE IV







HLA Class I/II Motifs/Supermotifs


TABLE IV (A): HLA Class I Supermotifs/Motifs












POSITION
POSITION



POSITION
3 (Primary
C Terminus



2 (Primary Anchor)
Anchor)
(Primary Anchor)





SUPERMOTIFS





A1
TILVMS

FWY


A2
LIVMATQ

IVMATL


A3
VSMATLI

RK


A24
YFWIVLMT

FIYWLM


B7
P

VILFMWYA


B27
RHK

FYLWMIVA


B44
ED

FWYLIMVA


B58
ATS

FWYLIVMA


B62
QLIVMP

FWYMIVLA


MOTIFS





A1
TSM

Y


A1

DEAS
Y


A2.1
LMVQIAT

VLIMAT


A3
LMVISATFCGD

KYRHFA


A11
VTMLISAGNCDF

KRYH


A24
YFWM

FLIW


A*3101
MVTALIS

RK


A*3301
MVALFIST

RK


A*6801
AVTMSLI

RK


B*0702
P

LMFWYAIV


B*3501
P

LMFWYIVA


B51
P

LIVFWYAM


B*5301
P

IMFWYALV


B*5401
P

ATIVLMFWY









Bolded residues are preferred, italicized residues are less preferred: A peptide is considered motif-bearing if it has primary anchors at each primary anchor position for a motif or supermotif as specified in the above table.







TABLE IV(B)







HLA Class II Supermotif









1
6
9





W, F, Y, V, .I, L
A, V, I, L, P, C, S, T
A, V, I, L, C, S, T, M, Y
















TABLE IV (C)







HLA Class II Motifs

















MOTIFS

1° anchor 1
2
3
4
5
1° anchor 6
7
8
9





DR4
preferred
FMYLIVW
M
T

I
VSTCPALIM
MH

MH



deleterious



W


R

WDE


DR1
preferred
MFLIVWY


PAMQ

VMATSPLIC
M

AVM



deleterious

C
CH
FD
CWD

GDE
D



DR7
preferred
MFLIVWY
M
W
A

IVMSACTPL
M

IV



deleterious

C

G


GRD
N
G


DR3
MOTIFS
1° anchor 1
2
3
1° anchor 4
5
1° anchor 6





motif a

LIVMFY


D







preferred












motif b

LIVMFAY


DNQEST

KRH





preferred












DR

MFLIVWY




VMSTACPLI





Supermotif





Italicized residues indicate less preferred or “tolerated” residues













TABLE IV (D)







HLA Class I Supermotifs










SUPER-

POSITION:


















MOTIFS

1
2
3
4
5
6
7
8
C-terminus





A1


1° Anchor






1° Anchor





TILVMS






FWY


A2


1° Anchor






1° Anchor





LIVMATQ






LIVMAT


A3
preferred

1° Anchor
YFW


YFW
YFW
P
1° Anchor





VSMATLI
(4/5)


(3/5)
(4/5)
(4/5)
RK



deleterious
DE (3/5);

DE










P (5/5)

(4/5)








A24


1° Anchor






1° Anchor





YFWIVLMT






FIYWLM


B7
preferred
FWY (5/5)
1° Anchor
FWY




FWY
1° Anchor




LIVM (3/5)
P
(4/5)




(3/5)
VILFMWYA



deleterious
DE (3/5);



DE
G
QN
DE





P (5/5); G (4/5);



(3/5)
(4/5)
(4/5)
(4/5)





A(3/5); QN(3/5)










B27


1° Anchor






1° Anchor





RIHK






FYLWMIVA


B44


1° Anchor






1° Anchor





ED






FWYLIMVA


B58


1° Anchor






1° Anchor





ATS






FWYLIVMA


B62


1° Anchor






1° Anchor





QLIVMP






FWYMIVLA





Italicized residues indicate less preferred or “tolerated” residues













TABLE IV (E)





HLA Class I Motifs

















POSITION:















1
2
3
4
5





A1
preferred
GFYW
1° Anchor
DEA
YFW



9-mer


STM






deleterious
DE

RHKLIVMP
A
G


A1
preferred
GRHK
ASTCLIVM
1° Anchor
GSTC



9-mer



DEAS





deleterious
A
RHKDEPYFW

DE
PQN


A1
preferred
YFW
1° Anchor
DEAQN
A
YFWQN


10-mer


STM






deleterious
GP

RHKGLIVM
DE
RHK


A1
preferred
YFW
STCLIVM
1° Anchor
A
YFW


10-mer



DEAS





deleterious
RHK
RHKDEPYFW


P


A2.1
preferred
YFW
1° Anchor
YFW
STC
YFW


9-mer


LMIVQAT






deleterious
DEP

DERKH




A2.1
preferred
AYFW
1° Anchor
LVIM
G



10-mer


LMIVQAT






deleterious
DEP

DE
RKHA
P


A3
preferred
RHK
1° Anchor
YFW
PRHKYFW
A





LMVISAT








FCGD






deleterious
DEP

DE




A11
preferred
A
1° Anchor
YFW
YFW
A





VTLMISA








GNCDF






deleterious
DEP






A24
preferred
YFWRHK
1° Anchor

STC



9-mer


YFWM






deleterious
DEG

DE
G
QNP


A24
preferred

1° Anchor

P
YFWP


10-mer


YFWM






deleterious


GDE
QN
RHK


A3101
preferred
RHK
1° Anchor
YFW
P






MVTALIS






deleterious
DEP

DE

ADE


A3301
preferred

1° Anchor
YFW







MVALFIST






deleterious
GP

DE




A6801
preferred
YFWSTC
1° Anchor


YFWLIVM





AVTMSLI






deleterious
GP

DEG

RHK


B0702
preferred
RHKFWY
1° Anchor
RHK

RHK





P






deleterious
DEQNP

DEP
DE
DE


B3501
preferred
FWYLIVM
1° Anchor
FWY







P






deleterious
AGP



G


B51
preferred
LIVMFWY
1° Anchor
FWY
STC
FWY





P






deleterious
AGPDER



DE




HKSTC






B5301
preferred
LIVMFWY
1° Anchor
FWY
STC
FWY





P






deleterious
AGPQN






B5401
preferred
FWY
1° Anchor
FWYLIVM

LIVM





P






deleterious
GPQNDE

GDESTC

RHKDE













POSITION:
















6
7
8
9
C-terminus





A1
preferred
P
DEQN
YFW
or C-terminus



9-mer




1° Anchor








Y




deleterious
A






A1
preferred
ASTC
LIVM
DE
1° Anchor



9-mer




Y




deleterious
RHK
PG
GP




A1
preferred

PASTC
GDE
P
1° Anchor


10-mer





Y



deleterious
QNA
RHKYFW
RHK
A



A1
preferred

PG
G
YFW
1° Anchor


10-mer





Y



deleterious
G

PRHK
QN



A2.1
preferred

A
P
1° Anchor



9-mer




VLIMAT




deleterious
RKH
DERKH





A2.1
preferred
G

FYWL

1° Anchor


10-mer



VIM

VLIMAT



deleterious

RKH
DERKH
RKH



A3
preferred
YFW

P
1° Anchor








KYRHFA




deleterious







A11
preferred
YFW
YFW
P
1° Anchor








KRYH




deleterious

A
G




A24
preferred

YFW
YFW
1° Anchor



9-mer




FLIW




deleterious
DERHK
G
AQN




A24
preferred

P


1° Anchor


10-mer





FLIW



deleterious
DE
A
QN
DEA



A3101
preferred
YFW
YFW
AP
1° Anchor








RK




deleterious
DE
DE
DE




A3301
preferred

AYFW

1° Anchor








RK




deleterious







A6801
preferred

YFW
P
1° Anchor








RK




deleterious


A




B0702
preferred
RHK
RHK
PA
1° Anchor








LMFWYAIV




deleterious
GDE
QN
DE




B3501
preferred

FWY

1° Anchor








LMFWYIVA




deleterious
G






B51
preferred

G
FWY
1° Anchor








LIVFWYAM




deleterious
G
DEQN
GDE




B5301
preferred

LIVMFWY
FWY
1° Anchor








IMFWYALV




deleterious
G
RHKQN
DE




B5401
preferred

ALIVM
FWYAP
1° Anchor








ATIVLMFWY




deleterious
DE
QNDGE
DE





Italicized residues indicate less preferred or “tolerated” residues. The information in this Table is specific for 9-mers unless otherwise specified.
















TABLE V





Pos
123456789
Score
SeqID















v.1-A1-9-mers: 184P1E2










99
SHEPLPLAY
112.500
46


373
NGELQDFPY
56.250
47


566
IIDIPQLFK
50.000
48


620
LLEPLGLHC
45.000
49


22
GVETLVDIY
45.000
50


348
IQDEMELGY
18.750
51


301
STLPPLEVY
12.500
52


218
GPEDVCEAY
11.250
53


285
FTDTVVFRV
6.250
54


120
DCDLNCEGR
5.000
55


271
LLDDSNEDF
5.000
56


62
RADTRRWRF
5.000
57


561
LAECDIIDI
4.500
58


36
GTEMFEVYG
4.500
59


563
ECDIIDIPQ
2.500
60


383
RILGPDFGY
2.500
61


386
GPDFGYVTR
2.500
62


490
LASPGACFK
2.000
63


73
TLEIIVVMN
1.800
64


409
NLEVSPPVV
1.800
65


176
DLEDMSVMV
1.800
66


43
YGTPGVDIY
1.250
67


362
KTLPVVFDS
1.250
68


517
VVDDEQVKT
1.000
69


454
KVQPPVELF
1.000
70


587
LVNMLVLGK
1.000
71


462
FVDWLAVGH
1.000
72


335
TICPQAENR
1.000
73


446
VRDFLHAQK
1.000
74


630
FIDDFTPYH
1.000
75


229
VLGQDKVSY
1.000
76


534
NKDLINYNK
1.000
77


612
CLEEKVRSL
0.900
78


222
VCEAYRHVL
0.900
79


351
EMELGYVQA
0.900
80


339
QAENRNDRW
0.900
81


323
VAELARKAG
0.900
82


10
SLEHPTSAV
0.900
83


124
NCEGRQDRN
0.900
84


243
HGDEERFFV
0.625
85


480
APDGKGFRM
0.625
86


584
FPDLVNMLV
0.625
87


198
KLVLHTSSY
0.500
88


319
FVDAVAELA
0.500
89


470
HVDEFLSFV
0.500
90


186
RTQGPAALF
0.500
91


47
GVDIYISPN
0.500
92


547
CIDWNREVL
0.500
93


166
NCDQHVHCL
0.500
94


202
HTSSYDAKR
0.500
95


574
KTERKKATA
0.450
96


393
TREPRDRSV
0.450
97


250
FVEGLSFPD
0.450
98


321
DAVAELARK
0.400
99


532
LSNKDLINY
0.375
100


174
LQDLEDMSV
0.375
101


412
VSPPVVANG
0.300
102


128
RQDRNFVDK
0.300
103


432
GNLPGSSGR
0.250
104


647
GTNVCRKPF
0.250
105


103
LPLAYAVLY
0.250
106


156
RDDPSCDVQ
0.250
107


478
VPAPDGKGF
0.250
108


641
HGEVHCGTN
0.225
109


236
SYEVPRLHG
0.225
110


253
GLSFPDAGF
0.200
111


302
TLPPLEVYV
0.200
112


645
HCGTNVCRK
0.200
113


411
EVSPPVVAN
0.200
114


527
SINQVLSNK
0.200
115


516
GVVDDEQVK
0.200
116


489
LLASPGACF
0.200
117


375
ELQDFPYKR
0.200
118


399
RSVSGLDSF
0.150
119


254
LSFPDAGFT
0.150
120


52
ISPNMERGR
0.150
121


31
GSVPEGTEM
0.150
122


261
FTGLISFHV
0.125
123


506
CGHGRALLF
0.125
124


634
FTPYHMLHG
0.125
125


259
AGFTGLISF
0.125
126


35
EGTEMFEVY
0.125
127


44
GTPGVDIYI
0.125
128


88
NDSHVQISY
0.125
129


256
FPDAGFTGL
0.125
130


177
LEDMSVMVL
0.125
131


84
SNDLNDSHV
0.125
132


193
LFDDHKLVL
0.125
133


87
LNDSHVQIS
0.125
134


33
VPEGTEMFE
0.113
135


433
NLPGSSGRR
0.100
136


610
CCCLEEKVR
0.100
137


51
YISPNMERG
0.100
138


354
LGYVQAPHK
0.100
139


466
LAVGHVDEF
0.100
140


241
RLHGDEERF
0.100
141


32
SVPEGTEMF
0.100
142


384
ILGPDFGYV
0.100
143


565
DIIDIPQLF
0.100
144


153
NCDRDDPSC
0.100
145







v.2-A1-9mers: 184P1E2










6
STLAPLEVY
1.250
146


7
TLAPLEVYV
0.200
147


9
APLEVYVCR
0.050
148


3
MTPSTLAPL
0.050
149


2
IMTPSTLAP
0.025
150


8
LAPLEVYVC
0.020
151


1
WIMTPSTLA
0.010
152


5
PSTLAPLEV
0.008
153


4
TPSTLAPLE
0.001
154







v.3-A1-9mers: 184P1E2










9
VPDGKGFRM
0.625
155


7
VPVPDGKGF
0.250
156


5
SFVPVPDGK
0.100
157


4
LSFVPVPDG
0.030
158


8
PVPDGKGFR
0.010
159


6
FVPVPDGKG
0.010
160


3
FLSFVPVPD
0.002
161


1
DEFLSFVPV
0.001
162


2
EFLSFVPVP
0.000
163



















TABLE VI





Pos
1234567890
Score
SeqID















v.1-A1-10mers: 184P1E2










87
LNDSHVQISY
31.250
164


319
FVDAVAELAR
25.000
165


547
CIDWNREVLK
20.000
166


373
NGELQDFPYK
9.000
167


124
NCEGRQDRNF
9.000
168


620
LLEPLGLHCT
9.000
169


218
GPEDVCEAYR
4.500
170


22
GVETLVDIYG
4.500
171


36
GTEMFEVYGT
4.500
172


73
TLEIIVVMNS
4.500
173


176
DLEDMSVMVL
4.500
174


98
SSHEPLPLAY
3.750
175


412
VSPPVVANGK
3.000
176


403
GLDSFGNLEV
2.500
177


347
WIQDEMELGY
2.500
178


566
IIDIPQLFKT
2.500
179


531
VLSNKDLINY
2.500
180


574
KTERKKATAF
2.250
181


10
SLEHPTSAVC
1.800
182


409
NLEVSPPVVA
1.800
183


250
FVEGLSFPDA
1.800
184


222
VCEAYRHVLG
1.800
185


323
VAELARKAGC
1.800
186


31
GSVPEGTEMF
1.500
187


628
CTFIDDFTPY
1.250
188


285
FTDTVVFRVA
1.250
189


33
VPEGTEMFEV
1.125
190


47
GVDIYISPNM
1.000
191


228
HVLGQDKVSY
1.000
192


565
DIIDIPQLFK
1.000
193


630
FIDDFTPYHM
1.000
194


586
DLVNMLVLGK
1.000
195


51
YISPNMERGR
1.000
196


563
ECDIIDIPQL
1.000
197


339
QAENRNDRWI
0.900
198


305
PLEVYVCRVR
0.900
199


99
SHEPLPLAYA
0.900
200


612
CLEEKVRSLL
0.900
201


254
LSFPDAGFTG
0.750
202


584
FPDLVNMLVL
0.625
203


372
RNGELQDFPY
0.625
204


490
LASPGACFKL
0.500
205


454
KVQPPVELFV
0.500
206


505
KCGHGRALLF
0.500
207


276
NEDFSASPIF
0.500
208


166
NCDQHVHCLQ
0.500
209


120
DCDLNCEGRQ
0.500
210


462
FVDWLAVGHV
0.500
211


517
VVDDEQVKTI
0.500
212


118
SLDCDLNCEG
0.500
213


114
CVDISLDCDL
0.500
214


160
SCDVQDNCDQ
0.500
215


49
DIYISPNMER
0.500
216


258
DAGFTGLISF
0.500
217


271
LLDDSNEDFS
0.500
218


301
STLPPLEVYV
0.500
219


177
LEDMSVMVLR
0.500
220


393
TREPRDRSVS
0.450
221


353
ELGYVQAPHK
0.400
222


526
ISINQVLSNK
0.300
223


475
LSFVPAPDGK
0.300
224


470
HVDEFLSFVP
0.250
225


534
NKDLINYNKF
0.250
226


334
LTICPQAENR
0.250
227


205
SYDAKRAQVF
0.250
228


478
VPAPDGKGFR
0.250
229


432
GNLPGSSGRR
0.250
230


641
HGEVHCGTNV
0.225
231


519
DDEQVKTISI
0.225
232


275
SNEDESASPI
0.225
233


489
LLASPGACFK
0.200
234


488
MLLASPGACF
0.200
235


494
GACFKLFQEK
0.200
236


643
EVHCGTNVCR
0.200
237


302
TLPPLEVYVC
0.200
238


384
ILGPDFGYVT
0.200
239


411
EVSPPVVANG
0.200
240


200
VLHTSSYDAK
0.200
241


491
ASPGACFKLF
0.150
242


112
LTCVDISLDC
0.125
243


21
AGVETLVDIY
0.125
244


580
ATAFFPDLVN
0.125
245


272
LDDSNEDFSA
0.125
246


256
FPDAGFTGLI
0.125
247


468
VGHVDEFLSF
0.125
248


194
FDDHKLVLHT
0.125
249


69
RFDATLEIIV
0.125
250


43
YGTPGVDIYI
0.125
251


442
VTQVVRDFLH
0.125
252


84
SNDLNDSHVQ
0.125
253


382
KRILGPDFGY
0.125
254


480
APDGKGFRML
0.125
255


217
CGPEDVCEAY
0.125
256


648
TNVCRKPFSF
0.125
257


609
GCCCLEEKVR
0.100
258


625
GLHCTFIDDF
0.100
259


335
TICPQAENRN
0.100
260


280
SASPIFTDTV
0.100
261


241
RLHGDEERFF
0.100
262


122
DLNCEGRQDR
0.100
263







v.2-A1-10mers: 184P1E2










9
LAPLEVYVCR
0.200
264


8
TLAPLEVYVC
0.200
265


3
IMTPSTLAPL
0.100
266


6
PSTLAPLEVY
0.075
267


7
STLAPLEVYV
0.050
268


5
TPSTLAPLEV
0.013
269


4
MTPSTLAPLE
0.005
270


2
WIMTPSTLAP
0.005
271


10
APLEVYVCRV
0.003
272


1
PWIMTPSTLA
0.000
273







v.3-A1-10mers: 184P1E2










5
LSFVPVPDGK
0.300
274


8
VPVPDGKGFR
0.250
275


10
VPDGKGFRML
0.125
276


7
FVPVPDGKGF
0.100
277


1
VDEFLSFVPV
0.090
278


4
FLSFVPVPDG
0.020
279


6
SFVPVPDGKG
0.005
280


9
PVPDGKGFRN
0.005
281


2
DEFLSFVPVP
0.000
282


3
EFLSFVPVPD
0.000
283



















TABLE VII





Pos
123456789
Score
SeqID















v.1-A2-9mers: 184P1E2










192
ALFDDHKLV
1055.104
284


302
TLPPLEVYV
382.536
285


384
ILGPDFGYV
160.627
286


111
YLTCVDISL
98.267
287


263
GLISFHVTL
81.177
288


455
VQPPVELFV
65.934
289


498
KLFQEKQKC
64.336
290


589
NMLVLGKHL
57.085
291


109
VLYLTCVDI
56.754
292


25
TLVDIYGSV
55.607
293


460
ELFVDWLAV
32.811
294


289
VVFRVAPWI
31.581
295


536
DLINYNKFV
28.690
296


79
VMNSPSNDL
26.228
297


261
FTGLISFHV
25.773
298


285
FTDTVVFRV
23.711
299


470
HVDEFLSFV
16.312
300


234
KVSYEVPRL
15.047
301


487
RMLLASPGA
13.276
302


655
FSFKWWNMV
13.137
303


264
LISFHVTLL
11.485
304


107
YAVLYLTCV
10.220
305


356
YVQAPHKTL
8.598
306


200
VLHTSSYDA
8.446
307


579
KATAFFPDL
7.498
308


408
GNLEVSPPV
6.887
309


1
MSLQRIVRV
6.568
310


40
FEVYGTPGV
6.221
311


19
CVAGVETLV
6.086
312


104
PLAYAVLYL
5.945
313


591
LVLGKHLGI
5.742
314


467
AVGHVDEFL
5.038
315


163
VQDNCDQHV
4.795
316


174
LQDLEDMSV
4.795
317


459
VELFVDWLA
4.557
318


10
SLEHPTSAV
4.451
319


488
MLLASPGAC
4.062
320


144
SGYGGILLV
3.342
321


86
DLNDSHVQI
3.208
322


309
YVCRVRNNT
2.999
323


231
GQDKVSYEV
2.821
324


612
CLEEKVRSL
2.579
325


312
RVRNNTCFV
2.544
326


529
NQVLSNKDL
2.166
327


315
NNTCFVDAV
1.944
328


409
NLEVSPPVV
1.825
329


517
VVDDEQVKT
1.818
330


580
ATAFFPDLV
1.799
331


530
QVLSNKDLI
1.655
332


630
FIDDFTPYH
1.616
333


176
DLEDMSVMV
1.369
334


442
VTQVVRDFL
1.359
335


642
GEVHCGTNV
1.352
336


105
LAYAVLYLT
1.295
337


623
PLGLHCTFI
1.262
338


37
TEMFEVYGT
1.233
339


298
MTPSTLPPL
1.160
340


191
AALFDDHKL
1.098
341


132
NFVDKRQWV
1.089
342


376
LQDFPYKRI
1.079
343


437
SSGRRVTQV
1.044
344


15
TSAVCVAGV
1.044
345


653
KPFSFKWWN
0.987
346


72
ATLEIIVVM
0.987
347


358
QAPHKTLPV
0.966
348


179
DMSVMVLRT
0.928
349


281
ASPIFTDTV
0.845
350


9
VSLEHPTSA
0.836
351


117
ISLDCDLNC
0.836
352


584
FPDLVNMLV
0.835
353


444
QVVRDFLHA
0.826
354


491
ASPGACFKL
0.809
355


243
HGDEERFFV
0.808
356


44
GTPGVDIYI
0.797
357


558
ELGLAECDI
0.782
358


510
RALLFQGVV
0.724
359


17
AVCVAGVET
0.652
360


583
FFPDLVNML
0.644
361


21
AGVETLVDI
0.642
362


184
VLRTQGPAA
0.642
363


621
LEPLGLHCT
0.567
364


221
DVCEAYRHV
0.561
365


616
KVRSLLEPL
0.560
366


620
LLEPLGLHC
0.541
367


385
LGPDFGYVT
0.528
368


402
SGLDSFGNL
0.523
369


553
EVLKRELGL
0.519
370


618
RSLLEPLGL
0.516
371


417
VANGKEYPL
0.504
372


505
KCGHGRALL
0.488
373


256
FPDAGFTGL
0.488
374


638
HMLHGEVHC
0.451
375


359
APHKTLPVV
0.428
376


524
KTISINQVL
0.426
377


463
VDWLAVGHV
0.408
378


294
APWIMTPST
0.404
379


34
PEGTEMFEV
0.397
380


319
FVDAVAELA
0.392
381


421
KEYPLGRIL
0.381
382


523
VKTISINQV
0.378
383







v.2-A2-9mers: 184P1E2










7
TLAPLEVYV
382.536
384


1
WIMTPSTLA
11.626
385


3
MTPSTLAPL
1.160
386


8
LAPLEVYVC
0.111
387


2
IMTPSTLAP
0.018
388


5
PSTLAPLEV
0.010
389


9
APLEVYVCR
0.002
390


6
STLAPLEVY
0.002
391


4
TPSTLAPLE
0.000
392







v.3-A2-9mers: 184P1E2










1
DEFLSFVPV
0.713
393


9
VPDGKGFRM
0.128
394


3
FLSFVPVPD
0.069
395


4
LSFVPVPDG
0.007
396


6
FVRVPDGKG
0.004
397


7
VPVPDGKGF
0.001
398


8
PVPDGKGFR
0.000
399


2
EFLSFVPVP
0.000
400


5
SFVPVPDGK
0.000
401



















TABLE VIII





Pos
1234567890
Score
SeqID















v.1-A2-10mers: 184P1E2










173
CLQDLEDMSV
285.163
402


192
ALFDDHKLVL
132.868
403


454
KVQPPVELFV
117.152
404


560
GLAECDIIDI
98.381
405


619
SLLEPLGLHC
75.365
406


297
IMTPSTLPPL
60.325
407


263
GLISFHVTLL
49.134
408


383
RILGPDFGYV
48.070
409


348
IQDEMELGYV
32.332
410


403
GLDSFGNLEV
27.821
411


2
SLQRIVRVSL
21.362
412


384
ILGPDFGYVT
19.737
413


301
STLPPLEVYV
19.658
414


571
QLFKTERKKA
18.382
415


590
MLVLGKHLGI
17.736
416


514
FQGVVDDEQV
15.895
417


459
VELFVDWLAV
14.461
418


433
NLPGSSGRRV
13.910
419


639
MLHGEVHCGT
12.668
420


357
VQAPHKTLPV
11.988
421


78
VVMNSPSNDL
11.757
422


9
VSLEHPTSAV
11.709
423


103
LPLAYAVLYL
11.096
424


242
LHGDEERFFV
10.739
425


469
GHVDEFLSFV
9.519
426


566
IIDIPQLFKT
9.295
427


131
RNFVDKRQWV
9.239
428


289
VVFRVAPWIM
8.408
429


125
CEGRQDRNFV
7.438
430


535
KDLINYNKFV
6.880
431


579
KATAFFPDLV
6.295
432


204
SSYDAKRAQV
5.957
433


517
VVDDEQVKTI
5.854
434


612
CLEEKVRSLL
5.605
435


653
KPFSFKWWNM
5.355
436


253
GLSFPDAGFT
5.328
437


630
FIDDFTPYHM
5.153
438


441
RVTQVVRDFL
5.038
439


288
TVVFRVAPWI
5.021
440


375
ELQDFPYKRI
4.812
441


302
TLPPLEVYVC
4.685
442


284
IFTDTVVFRV
4.491
443


191
AALFDDHKLV
3.574
444


108
AVLYLTCVDI
3.378
445


462
FVDWLAVGHV
3.348
446


416
VVANGKEYPL
3.178
447


230
LGQDKVSYEV
3.165
448


490
LASPGACFKL
2.925
449


408
GNLEVSPPVV
2.824
450


184
VLRTQGPAAL
2.760
451


183
MVLRTQGPAA
2.734
452


199
LVLHTSSYDA
2.734
453


516
GVVDDEQVKT
2.673
454


11
LEHPTSAVCV
2.299
455


421
KEYPLGRILI
2.272
456


598
GIPKPFGPII
2.235
457


17
AVCVAGVETL
2.107
458


407
FGNLEVSPPV
2.088
459


309
YVCRVRNNTC
2.000
460


262
TGLISFHVTL
1.961
461


314
RNNTCFVDAV
1.944
462


162
DVQDNCDQHV
1.871
463


620
LLEPLGLHCT
1.865
464


280
SASPIFTDTV
1.799
465


487
RMLLASPGAC
1.726
466


43
YGTPGVDIYI
1.723
467


325
ELARKAGCKL
1.602
468


538
INYNKFVQSC
1.542
469


175
QDLEDMSVMV
1.465
470


466
LAVGHVDEFL
1.359
471


317
TCFVDAVAEL
1.187
472


294
APWIMTPSTL
1.157
473


436
GSSGRRVTQV
1.044
474


611
CCLEEKVRSL
0.973
475


601
KPFGPIINGC
0.966
476


498
KLFQEKQKCG
0.965
477


443
TQVVRDFLHA
0.957
478


583
FFPDLVNMLV
0.911
479


358
QAPHKTLPVV
0.911
480


588
VNMLVLGKHL
0.877
481


182
VMVLRTQGPA
0.793
482


18
VCVAGVETLV
0.772
483


557
RELGLAECDI
0.751
484


578
KKATAFFPDL
0.750
485


114
CVDISLDCDL
0.748
486


33
VPEGTEMFEV
0.659
487


581
TAFFPDLVNM
0.587
488


20
VAGVETLVDI
0.567
489


105
LAYAVLYLTC
0.563
490


271
LLDDSNEDFS
0.560
491


270
TLLDDSNEDF
0.554
492


529
NQVLSNKDLI
0.518
493


522
QVKTISINQV
0.490
494


488
MLLASPGACF
0.469
495


299
TPSTLPPLEV
0.454
496


8
RVSLEHPTSA
0.435
497


279
FSASPIFTDT
0.425
498


70
FDATLEIIVV
0.419
499


552
REVLKRELGL
0.415
500


176
DLEDMSVMVL
0.396
501







v.2-A2-10mers: 184P1e2










3
IMTPSTLAPL
60.325
502


7
STLAPLEVYV
19.658
503


10
APLEVYVCRV
15.841
504


8
TLAPLEVYVC
4.685
505


5
TPSTLAPLEV
0.454
506


2
WIMTPSTLAP
0.035
507


9
LAPLEVYVCR
0.001
508


4
MTPSTLAPLE
0.000
509


1
PWIMTPSTLA
0.000
510


6
PSTLAPLEVY
0.000
511







v.3-A2-10mers: 184P1E2










4
FLSFVPVPDG
0.788
512


9
PVPDGKGFRM
0.227
513


10
VPDGKGFRML
0.036
514


7
FVPVPDGKGF
0.030
515


1
VDEFLSFVPV
0.020
516


5
LSFVPVPDGK
0.000
517


2
DEFLSFVPVP
0.000
518


8
VPVPDGKGFR
0.000
519


6
SFVPVPDGKG
0.000
520


3
EFLSFVPVPD
0.000
521



















TABLE IX





Pos
123456789
Score
SeqID















v.1-A3-9mers: 184P1E2










198
KLVLHTSSY
18.000
522


587
LVNMLVLGK
12.000
523


375
ELQDFPYKR
10.800
524


571
QLFKTERKK
10.000
525


516
GVVDDEQVK
9.000
526


253
GLSFPDAGF
9.000
527


263
GLISFHVTL
8.100
528


566
IIDIPQLFK
6.000
529


527
SINQVLSNK
4.500
530


383
RILGPDFGY
4.050
531


454
KVQPPVELF
4.050
532


229
VLGQDKVSY
4.000
533


22
GVETLVDIY
3.600
534


495
ACFKLFQEK
3.000
535


498
KLFQEKQKC
3.000
536


241
RLHGDEERF
3.000
537


489
LLASPGACF
3.000
538


109
VLYLTCVDI
3.000
539


148
GILLVNCDR
2.700
540


271
LLDDSNEDF
2.000
541


433
NLPGSSGRR
1.800
542


111
YLTCVDISL
1.800
543


128
RQDRNFVDK
1.800
544


325
ELARKAGCK
1.800
545


301
STLPPLEVY
1.012
546


79
VMNSPSNDL
0.900
547


650
VCRKPFSFK
0.900
548


374
GELQDFPYK
0.810
549


645
HCGTNVCRK
0.600
550


620
LLEPLGLHC
0.600
551


200
VLHTSSYDA
0.600
552


335
TICPQAENR
0.600
553


649
NVCRKPFSF
0.600
554


460
ELFVDWLAV
0.600
555


202
HTSSYDAKR
0.600
556


544
VQSCIDWNR
0.540
557


234
KVSYEVPRL
0.540
558


192
ALFDDHKLV
0.500
559


289
VVFRVAPWI
0.450
560


589
NMLVLGKHL
0.450
561


44
GTPGVDIYI
0.405
562


625
GLHCTFIDD
0.360
563


348
IQDEMELGY
0.360
564


104
PLAYAVLYL
0.360
565


619
SLLEPLGLH
0.304
566


10
SLEHPTSAV
0.300
567


302
TLPPLEVYV
0.300
568


490
LASPGACFK
0.300
569


512
LLFQGVVDD
0.300
570


487
RMLLASPGA
0.300
571


638
HMLHGEVHC
0.300
572


32
SVPEGTEMF
0.300
573


413
SPPVVANGK
0.300
574


384
ILGPDFGYV
0.270
575


612
CLEEKVRSL
0.270
576


616
KVRSLLEPL
0.270
577


591
LVLGKHLGI
0.270
578


598
GIPKPFGPI
0.243
579


186
RTQGPAALF
0.225
580


524
KTISINQVL
0.203
581


184
VLRTQGPAA
0.200
582


409
NLEVSPPVV
0.200
583


55
NMERGRERA
0.200
584


362
KTLPVVFDS
0.182
585


103
LPLAYAVLY
0.180
586


558
ELGLAECDI
0.180
587


444
QVVRDFLHA
0.180
588


218
GPEDVCEAY
0.180
589


264
LISFHVTLL
0.180
590


86
DLNDSHVQI
0.180
591


179
DMSVMVLRT
0.180
592


305
PLEVYVCRV
0.180
593


560
GLAECDIID
0.180
594


364
LPVVFDSPR
0.180
595


403
GLDSFGNLE
0.180
596


579
KATAFFPDL
0.162
597


647
GTNVCRKPF
0.150
598


470
HVDEFLSFV
0.135
599


536
DLINYNKFV
0.135
600


25
TLVDIYGSV
0.135
601


565
DIIDIPQLF
0.135
602


321
DAVAELARK
0.135
603


285
FTDTVVFRV
0.135
604


466
LAVGHVDEF
0.135
605


386
GPDFGYVTR
0.120
606


569
IPQLFKTER
0.120
607


354
LGYVQAPHK
0.100
608


548
IDWNREVLK
0.100
609


283
PIFTDTVVF
0.100
610


570
PQLFKTERK
0.090
611


534
NKDLINYNK
0.090
612


428
ILIGGNLPG
0.090
613


288
TVVFRVAPW
0.090
614


363
TLPVVFDSP
0.090
615


467
AVGHVDEFL
0.090
616


353
ELGYVQAPH
0.090
617


441
RVTQVVRDF
0.090
618


5
RIVRVSLEH
0.090
619


530
QVLSNKDLI
0.090
620


554
VLKRELGLA
0.090
621







v.2-A3-9mers: 184P1E2










6
STLAPLEVY
0.675
622


7
TLAPLEVYV
0.300
623


9
APLEVYVCR
0.135
624


2
IMTPSTLAP
0.060
625


1
WIMTPSTLA
0.045
626


3
MTPSTLAPL
0.045
627


8
LAPLEVYVC
0.018
628


4
TPSTLAPLE
0.000
629


5
PSTLAPLEV
0.000
630







v.3-A3-9mers- 184P1E2










5
SFVPVPDGK
0.068
631


3
FLSFVPVPD
0.030
632


7
VPVPDGKGF
0.022
633


8
PVPDGKGFR
0.012
634


9
VPDGKGFRM
0.009
635


1
DEFLSFVPV
0.003
636


4
LSFVPVPDG
0.002
637


6
FVPVPDGKG
0.001
638


2
EFLSFVPVP
0.000
639



















TABLE X





Pos
1234567890
Score
SeqID















v.1-A3-10mers: 184P1E2










586
DLVNMLVLGK
54.000
640


592
VLGKHLGIPK
40.000
641


625
GLHCTFIDDF
27.000
642


489
LLASPGACFK
20.000
643


200
VLHTSSYDAK
20.000
644


363
TLPVVFDSPR
12.000
645


531
VLSNKDLINY
12.000
646


649
NVCRKPFSFK
9.000
647


560
GLAECDIIDI
8.100
648


263
GLISFHVTLL
8.100
649


192
ALFDDHKLVL
6.000
650


465
WLAVGHVDEF
6.000
651


353
ELGYVQAPHK
6.000
652


488
MLLASPGACF
4.500
653


270
TLLDDSNEDF
3.000
654


628
CTFIDDFTPY
3.000
655


565
DIIDIPQLFK
2.700
656


302
TLPPLEVYVC
2.700
657


445
VVRDFLHAQK
2.000
658


547
CIDWNREVLK
2.000
659


590
MLVLGKHLGI
1.800
660


189
GPAALFDDHK
1.800
661


49
DIYISPNMER
1.800
662


494
GACFKLFQEK
1.800
663


543
FVQSCIDWNR
1.800
664


612
CLEEKVRSLL
1.350
665


297
IMTPSTLPPL
1.350
666


384
ILGPDFGYVT
1.350
667


122
DLNCEGRQDR
1.200
668


102
PLPLAYAVLY
1.200
669


403
GLDSFGNLEV
1.200
670


176
DLEDMSVMVL
1.080
671


128
RQDRNFVDKR
1.080
672


619
SLLEPLGLHC
0.900
673


228
HVLGQDKVSY
0.900
674


2
SLQRIVRVSL
0.900
675


347
WIQDEMELGY
0.800
676


319
FVDAVAELAR
0.800
677


184
VLRTQGPAAL
0.600
678


283
PIFTDTVVFR
0.600
679


109
VLYLTCVDIS
0.600
680


137
RQWVWGPSGY
0.600
681


598
GIPKPFGPII
0.540
682


475
LSFVPAPDGK
0.500
683


571
QLFKTERKKA
0.500
684


374
GELQDFPYKR
0.486
685


512
LLFQGVVDDE
0.450
686


173
CLQDLEDMSV
0.400
687


568
DIPQLFKTER
0.360
688


73
TLEIIVVMNS
0.360
689


526
ISINQVLSNK
0.338
690


289
VVFRVAPWIM
0.300
691


241
RLHGDEERFF
0.300
692


334
LTICPQAENR
0.300
693


574
KTERKKATAF
0.300
694


409
NLEVSPPVVA
0.300
695


454
KVQPPVELFV
0.270
696


639
MLHGEVHCGT
0.225
697


412
VSPPVVANGK
0.225
698


620
LLEPLGLHCT
0.225
699


41
EVYGTPGVDI
0.203
700


569
IPQLFKTERK
0.200
701


10
SLEHPTSAVC
0.200
702


127
GRQDRNFVDK
0.180
703


533
SNKDLINYNK
0.180
704


558
ELGLAECDII
0.180
705


325
ELARKAGCKL
0.180
706


416
VVANGKEYPL
0.180
707


17
AVCVAGVETL
0.180
708


498
KLFQEKQKCG
0.150
709


460
ELFVDWLAVG
0.135
710


38
EMFEVYGTPG
0.135
711


375
ELQDFPYKRI
0.135
712


36
GTEMFEVYGT
0.135
713


78
VVMNSPSNDL
0.135
714


288
TVVFRVAPWI
0.135
715


428
ILIGGNLPGS
0.135
716


548
IDWNREVLKR
0.120
717


218
GPEDVCEAYR
0.120
718


643
EVHCGTNVCR
0.120
719


505
KCGHGRALLF
0.120
720


601
KPFGPIINGC
0.101
721


477
FVPAPDGKGF
0.100
722


644
VHCGTNVCRK
0.090
723


333
KLTICPQAEN
0.090
724


47
GVDIYISPNM
0.090
725


517
VVDDEQVKTI
0.090
726


653
KPFSFKWWNM
0.090
727


182
VMVLRTQGPA
0.090
728


317
TCFVDAVAEL
0.090
729


25
TLVDIYGSVP
0.090
730


198
KLVLHTSSYD
0.090
731


441
RVTQVVRDFL
0.090
732


487
RMLLASPGAC
0.090
733


250
FVEGLSFPDA
0.090
734


199
LVLHTSSYDA
0.090
735


511
ALLFQGVVDD
0.090
736


253
GLSFPDAGFT
0.090
737


108
AVLYLTCVDI
0.090
738


638
HMLHGEVHCG
0.090
739







v.2-A3-10mers: 184P1E2










8
TLAPLEVYVC
2.700
740


3
IMTPSTLAPL
1.350
741


9
LAPLEVYVCR
0.060
742


10
APLEVYVCRV
0.041
743


7
STLAPLEVYV
0.015
744


2
WIMTPSTLAP
0.006
745


6
PSTLAPLEVY
0.005
746


5
TPSTLAPLEV
0.004
747


4
MTPSTLAPLE
0.001
748


1
PWIMTPSTLA
0.000
749







v.3-A3-10mers: 184P1E2










5
LSFVPVPDGK
0.750
750


7
FVPVPDGKGF
0.100
751


4
FLSFVPVPDG
0.090
752


8
VPVPDGKGFR
0.027
753


9
PVPDGKGFRM
0.006
754


10
VPDGKGFRML
0.003
755


2
DEFLSFVPVP
0.001
756


1
VDEFLSFVPV
0.001
757


6
SFVPVPDGKG
0.000
758


3
EFLSFVPVPD
0.000
759



















TABLE XI





Pos
123456789
Score
SeqID















v.1-A11-9mers: 184P1E2










516
GVVDDEQVK
9.000
760


587
LVNMLVLGK
4.000
761


128
RQDRNFVDK
1.800
762


566
IIDIPQLFK
0.800
763


495
ACFKLFQEK
0.400
764


527
SINQVLSNK
0.400
765


148
GILLVNCDR
0.360
766


476
SFVPAPDGK
0.300
767


374
GELQDFPYK
0.270
768


50
IYISPNMER
0.240
769


544
VQSCIDWNR
0.240
770


202
HTSSYDAKR
0.200
771


490
LASPGACFK
0.200
772


645
HCGTNVCRK
0.200
773


650
VCRKPFSFK
0.200
774


413
SPPVVANGK
0.200
775


325
ELARKAGCK
0.120
776


386
GPDFGYVTR
0.120
777


570
PQLFKTERK
0.090
778


321
DAVAELARK
0.090
779


571
QLFKTERKK
0.080
780


335
TICPQAENR
0.080
781


433
NLPGSSGRR
0.080
782


22
GVETLVDIY
0.060
783


312
RVRNNTCFV
0.060
784


649
NVCRKPFSF
0.060
785


444
QVVRDFLHA
0.060
786


616
KVRSLLEPL
0.060
787


364
LPVVFDSPR
0.060
788


44
GTPGVDIYI
0.060
789


454
KVQPPVELF
0.060
790


591
LVLGKHLGI
0.060
791


234
KVSYEVPRL
0.060
792


383
RILGPDFGY
0.054
793


375
ELQDFPYKR
0.048
794


524
KTISINQVL
0.045
795


354
LGYVQAPHK
0.040
796


284
IFTDTVVFR
0.040
797


391
YVTREPRDR
0.040
798


289
VVFRVAPWI
0.040
799


593
LGKHLGIPK
0.040
800


133
FVDKRQWVW
0.040
801


548
IDWNREVLK
0.040
802


569
IPQLFKTER
0.040
803


534
NKDLINYNK
0.040
804


432
GNLPGSSGR
0.036
805


231
GQDKVSYEV
0.036
806


5
RIVRVSLEH
0.036
807


288
TVVFRVAPW
0.030
808


530
QVLSNKDLI
0.030
809


285
FTDTVVFRV
0.030
810


183
MVLRTQGPA
0.030
811


186
RTQGPAALF
0.030
812


574
KTERKKATA
0.030
813


261
FTGLISFHV
0.030
814


443
TQVVRDFLH
0.027
815


248
RFFVEGLSF
0.024
816


201
LHTSSYDAK
0.020
817


462
FVDWLAVGH
0.020
818


467
AVGHVDEFL
0.020
819


319
FVDAVAELA
0.020
820


226
YRHVLGQDK
0.020
821


32
SVPEGTEMF
0.020
822


19
CVAGVETLV
0.020
823


470
HVDEFLSFV
0.020
824


610
CCCLEEKVR
0.020
825


446
VRDFLHAQK
0.020
826


608
NGCCCLEEK
0.020
827


190
PAALFDDHK
0.020
828


503
KQKCGHGRA
0.018
829


487
RMLLASPGA
0.018
830


263
GLISFHVTL
0.018
831


198
KLVLHTSSY
0.018
832


542
KFVQSCIDW
0.018
833


553
EVLKRELGL
0.018
834


497
FKLFQEKQK
0.015
835


647
GTNVCRKPF
0.015
836


301
STLPPLEVY
0.015
837


72
ATLEIIVVM
0.015
838


362
KTLPVVFDS
0.013
839


59
GRERADTRR
0.012
840


338
PQAENRNDR
0.012
841


58
RGRERADTR
0.012
842


439
GRRVTQVVR
0.012
843


348
IQDEMELGY
0.012
844


120
DCDLNCEGR
0.012
845


241
RLHGDEERF
0.012
846


598
GIPKPFGPI
0.012
847


62
RADTRRWRF
0.012
848


174
LQDLEDMSV
0.012
849


292
RVAPWIMTP
0.012
850


253
GLSFPDAGF
0.012
851


455
VQPPVELFV
0.012
852


298
MTPSTLPPL
0.010
853


580
ATAFFPDLV
0.010
854


316
NTCFVDAVA
0.010
855


356
YVQAPHKTL
0.010
856


442
VTQVVRDFL
0.010
857


510
RALLFQGVV
0.009
858


64
DTRRWRFDA
0.009
859







v.2-A11-9mers: 184P1E2










9
APLEVYVCR
0.060
860


6
STLAPLEVY
0.015
861


3
MTPSTLAPL
0.010
862


1
WIMTPSTLA
0.008
863


7
TLAPLEVYV
0.004
864


2
IMTPSTLAP
0.001
865


8
LAPLEVYVC
0.000
866


4
TPSTLAPLE
0.000
867


5
PSTLAPLEV
0.000
868







v.3-A11-9mers: 184P1E2










5
SFVPVPDGK
0.300
869


8
PVPDGKGFR
0.040
870


9
VPDGKGFRM
0.006
871


6
FVPVPDGKG
0.002
872


7
VPVPDGKGF
0.002
873


3
FLSFVPVPD
0.000
874


1
DEFLSFVPV
0.000
875


2
EFLSFVPVP
0.000
876


4
LSFVPVPDG
0.000
877



















TABLE XII





Pos
1234567890
Score
SeqID















v.1-A11-10mers: 184P1E2










445
VVRDFLHAQK
2.000
878


649
NVCRKPFSFK
2.000
879


592
VLGKHLGIPK
0.800
880


543
FVQSCIDWNR
0.800
881


319
FVDAVAELAR
0.800
882


189
GPAALFDDHK
0.600
883


494
GACFKLFQEK
0.600
884


200
VLHTSSYDAK
0.400
885


489
LLASPGACFK
0.400
886


225
AYRHVLGQDK
0.400
887


547
CIDWNREVLK
0.400
888


128
RQDRNFVDKR
0.360
889


565
DIIDIPQLFK
0.360
890


586
DLVNMLVLGK
0.360
891


334
LTICPQAENR
0.300
892


569
IPQLFKTERK
0.200
893


218
GPEDVCEAYR
0.120
894


454
KVQPPVELFV
0.120
895


643
EVHCGTNVCR
0.120
896


353
ELGYVQAPHK
0.120
897


374
GELQDFPYKR
0.108
898


496
CFKLFQEKQK
0.100
899


49
DIYISPNMER
0.096
900


324
AELARKAGCK
0.090
901


363
TLPVVFDSPR
0.080
902


289
VVFRVAPWIM
0.080
903


533
SNKDLINYNK
0.080
904


127
GRQDRNFVDK
0.060
905


609
GCCCLEEKVR
0.060
906


441
RVTQVVRDFL
0.060
907


199
LVLHTSSYDA
0.060
908


8
RVSLEHPTSA
0.060
909


47
GVDIYISPNM
0.060
910


78
VVMNSPSNDL
0.040
911


478
VPAPDGKGFR
0.040
912


416
VVANGKEYPL
0.040
913


337
CPQAENRNDR
0.040
914


607
INGCCCLEEK
0.040
915


239
VPRLHGDEER
0.040
916


303
LPPLEVYVCR
0.040
917


475
LSFVPAPDGK
0.040
918


432
GNLPGSSGRR
0.036
919


390
GYVTREPRDR
0.036
920


137
RQWVWGPSGY
0.036
921


526
ISINQVLSNK
0.030
922


515
QGVVDDEQVK
0.030
923


228
HVLGQDKVSY
0.030
924


108
AVLYLTCVDI
0.030
925


442
VTQVVRDFLH
0.030
926


574
KTERKKATAF
0.030
927


288
TVVFRVAPWI
0.030
928


183
MVLRTQGPAA
0.030
929


568
DIPQLFKTER
0.024
930


653
KPFSFKWWNM
0.024
931


122
DLNCEGRQDR
0.024
932


560
GLAECDIIDI
0.024
933


403
GLDSFGNLEV
0.024
934


628
CTFIDDFTPY
0.020
935


522
QVKTISINQV
0.020
936


412
VSPPVVANGK
0.020
937


250
FVEGLSFPDA
0.020
938


17
AVCVAGVETL
0.020
939


114
CVDISLDCDL
0.020
940


644
VHCGTNVCRK
0.020
941


462
FVDWLAVGHV
0.020
942


320
VDAVAELARK
0.020
943


91
HVQISYHSSH
0.020
944


587
LVNMLVLGKH
0.020
945


373
NGELQDFPYK
0.020
946


147
GGILLVNCDR
0.018
947


443
TQVVRDFLHA
0.018
948


383
RILGPDFGYV
0.018
949


263
GLISFHVTLL
0.018
950


260
GFTGLISFHV
0.018
951


312
RVRNNTCFVD
0.018
952


548
IDWNREVLKR
0.016
953


418
ANGKEYPLGR
0.016
954


192
ALFDDHKLVL
0.016
955


283
PIFTDTVVFR
0.016
956


301
STLPPLEVYV
0.015
957


431
GGNLPGSSGR
0.012
958


58
RGRERADTRR
0.012
959


501
QEKQKCGHGR
0.012
960


598
GIPKPFGPII
0.012
961


142
GPSGYGGILL
0.012
962


590
MLVLGKHLGI
0.012
963


69
RFDATLEIIV
0.012
964


357
VQAPHKTLPV
0.012
965


625
GLHCTFIDDF
0.012
966


41
EVYGTPGVDI
0.012
967


110
LYLTCVDISL
0.012
968


388
DFGYVTREPR
0.012
969


177
LEDMSVMVLR
0.012
970


22
GVETLVDIYG
0.012
971


505
KCGHGRALLF
0.012
972


477
FVPAPDGKGF
0.010
973


517
VVDDEQVKTI
0.010
974


570
PQLFKTERKK
0.009
975


516
GVVDDEQVKT
0.009
976


529
NQVLSNKDLI
0.009
977







v.2-A11-10mers: 184P1E2










9
LAPLEVYVCR
0.040
978


7
STLAPLEVYV
0.015
979


5
TPSTLAPLEV
0.004
980


3
TMTPSTLAPL
0.004
981


10
APLEVYVCRV
0.003
982


2
WIMTPSTLAP
0.002
983


4
MTPSTLAPLE
0.001
984


8
TLAPLEVYVC
0.001
985


1
PWIMTPSTLA
0.000
986


6
PSTLAPLEVY
0.000
987







v.3-A11-10mers: 184P1E2










8
VPVPDGKGFR
0.060
988


5
LSFVPVPDGK
0.040
989


7
FVPVPDGKGF
0.010
990


9
PVPDGKGFRM
0.006
991


4
FLSFVPVPDG
0.000
992


6
SFVPVPDGKG
0.000
993


10
VPDGKGFRML
0.000
994


1
VDEFLSFVPV
0.000
995


3
EFLSFVPVPD
0.000
996


2
DEFLSFVPVP
0.000
997



















TABLE XIII





Pos
123456789
Score
SeqID















v.1-A24-9mers: 184P1E2










95
SYHSSHEPL
200.000
998


422
EYPLGRILI
75.000
999


583
FFPDLVNML
51.840
1000


42
VYGTPGVDI
50.000
1001


318
CFVDAVAEL
39.600
1002


193
LFDDHKLVL
24.000
1003


524
KTISINQVL
20.160
1004


248
RFFVEGLSF
20.000
1005


346
RWIQDEMEL
13.200
1006


618
RSLLEPLGL
12.000
1007


69
RFDATLEII
10.000
1008


579
KATAFFPDL
9.600
1009


616
KVRSLLEPL
9.600
1010


539
NYNKFVQSC
9.000
1011


402
SGLDSFGNL
8.640
1012


442
VTQVVRDFL
8.400
1013


355
GYVQAPHKT
8.250
1014


505
KCGHGRALL
8.000
1015


234
KVSYEVPRL
8.000
1016


110
LYLTCVDIS
7.500
1017


308
VYVCRVRNN
7.500
1018


589
NMLVLGKHL
7.200
1019


454
KVQPPVELF
7.200
1020


79
VMNSPSNDL
7.200
1021


612
CLEEKVRSL
7.200
1022


145
GYGGILLVN
7.000
1023


29
IYGSVPEGT
7.000
1024


191
AALFDDHKL
6.600
1025


491
ASPGACFKL
6.600
1026


565
DIIDIPQLF
6.048
1027


298
MTPSTLPPL
6.000
1028


417
VANGKEYPL
6.000
1029


553
EVLKRELGL
6.000
1030


222
VCEAYRHVL
6.000
1031


529
NQVLSNKDL
6.000
1032


186
RTQGPAALF
6.000
1033


263
GLISFHVTL
6.000
1034


399
RSVSGLDSF
6.000
1035


18
VCVAGVETL
6.000
1036


356
YVQAPHKTL
6.000
1037


106
AYAVLYLTC
6.000
1038


111
YLTCVDISL
5.600
1039


467
AVGHVDEFL
5.600
1040


441
RVTQVVRDF
5.600
1041


3
LQRIVRVSL
5.600
1042


205
SYDAKRAQV
5.000
1043


256
FPDAGFTGL
4.800
1044


326
LARKAGCKL
4.400
1045


482
DGKGFRMLL
4.000
1046


62
RADTRRWRF
4.000
1047


264
LISFHVTLL
4.000
1048


547
CIDWNREVL
4.000
1049


97
HSSHEPLPL
4.000
1050


241
RLHGDEERF
4.000
1051


142
GPSGYGGIL
4.000
1052


166
NCDQHVHCL
4.000
1053


32
SVPEGTEMF
3.600
1054


622
EPLGLHCTF
3.600
1055


466
LAVGHVDEF
3.300
1056


647
GTNVCRKPF
3.000
1057


582
AFFPDLVNM
3.000
1058


290
VFRVAPWIM
2.500
1059


478
VPAPDGKGF
2.400
1060


271
LLDDSNEDF
2.400
1061


492
SPGACFKLF
2.400
1062


44
GTPGVDIYI
2.100
1063


649
NVCRKPFSF
2.000
1064


506
CGHGRALLF
2.000
1065


489
LLASPGACF
2.000
1066


253
GLSFPDAGF
2.000
1067


259
AGFTGLISF
2.000
1068


598
GIPKPFGPI
1.800
1069


86
DLNDSHVQI
1.800
1070


21
AGVETLVDI
1.800
1071


141
WGPSGYGGI
1.500
1072


561
LAECDIIDI
1.500
1073


530
QVLSNKDLI
1.500
1074


559
LGLAECDII
1.500
1075


591
LVLGKHLGI
1.500
1076


542
KFVQSCIDW
1.500
1077


289
VVFRVAPWI
1.400
1078


540
YNKFVQSCI
1.400
1079


72
ATLEIIVVM
1.260
1080


599
IPKPFGPII
1.200
1081


421
KEYPLGRIL
1.152
1082


132
NFVDKRQWV
1.080
1083


629
TFIDDFTPY
1.080
1084


558
ELGLAECDI
1.000
1085


376
LQDFPYKRI
1.000
1086


109
VLYLTCVDI
1.000
1087


31
GSVPEGTEM
0.990
1088


613
LEEKVRSLL
0.840
1089


595
KHLGIPKPF
0.840
1090


426
GRILIGGNL
0.840
1091


50
IYISPNMER
0.825
1092


66
RRWRFDATL
0.800
1093


551
NREVLKREL
0.792
1094


535
KDLINYNKF
0.792
1095


390
GYVTREPRD
0.750
1096


236
SYEVPRLHG
0.750
1097







v.2-A24-9mers: 184P1E2










3
MTPSTLAPL
6.000
1098


1
WIMTPSTLA
0.150
1099


8
LAPLEVYVC
0.150
1100


6
STLAPLEVY
0.150
1101


7
TLAPLEVYV
0.144
1102


9
APLEVYVCR
0.025
1103


4
TPSTLAPLE
0.012
1104


2
IMTPSTLAP
0.012
1105


5
PSTLAPLEV
0.011
1106







v.3-A24-9mers: 184P1E2










7
VPVPDGKGF
3.600
1107


9
VPDGKGFRM
0.500
1108


5
SFVPVPDGK
0.126
1109


2
EFLSFVPVP
0.075
1110


6
FVPVPDGKG
0.017
1111


3
FLSFVPVPD
0.014
1112


1
DEFLSFVPV
0.012
1113


4
LSFVPVPDG
0.010
1114


8
PVPDGKGFR
0.002
1115



















TABLE XIV





Pos
1234567890
Score
SeqID















v.1-A24-10mers: 184P1E2










110
LYLTCVDISL
420.000
1116


355
GYVQAPHKTL
300.000
1117


205
SYDAKRAQVF
120.000
1118


539
NYNKFVQSCI
105.000
1119


255
SFPDAGFTGL
43.200
1120


582
AFFPDLVNML
34.560
1121


367
VFDSPRNGEL
22.000
1122


308
VYVCRVRNNT
12.600
1123


380
PYKRILGPDF
12.000
1124


441
RVTQVVRDFL
11.200
1125


612
CLEEKVRSLL
10.080
1126


466
LAVGHVDEFL
8.400
1127


2
SLQRIVRVSL
8.400
1128


503
KQKCGHGRAL
8.000
1129


611
CCLEEKVRSL
7.200
1130


101
EPLPLAYAVL
7.200
1131


588
VNMLVLGKHL
7.200
1132


546
SCIDWNREVL
7.200
1133


176
DLEDMSVMVL
7.200
1134


78
VVMNSPSNDL
7.200
1135


550
WNREVLKREL
6.336
1136


263
GLISFHVTLL
6.000
1137


528
INQVLSNKDL
6.000
1138


141
WGPSGYGGIL
6.000
1139


574
KTERKKATAF
6.000
1140


262
TGLISFHVTL
6.000
1141


604
GPIINGCCCL
6.000
1142


103
LPLAYAVLYL
6.000
1143


145
GYGGILLVNC
6.000
1144


425
LGRILIGGNL
5.600
1145


490
LASPGACFKL
5.280
1146


106
AYAVLYLTCV
5.000
1147


42
VYGTPGVDIY
5.000
1148


114
CVDISLDCDL
4.800
1149


297
IMTPSTLPPL
4.800
1150


401
VSGLDSFGNL
4.800
1151


221
DVCEAYRHVL
4.800
1152


165
DNCDQHVHCL
4.800
1153


192
ALFDDHKLVL
4.800
1154


395
EPRDRSVSGL
4.800
1155


480
APDGKGFRML
4.800
1156


168
DQHVHCLQDL
4.800
1157


325
ELARKAGCKL
4.400
1158


452
AQKVQPPVEL
4.400
1159


317
TCFVDAVAEL
4.400
1160


376
LQDFPYKRIL
4.000
1161


584
FPDLVNMLVL
4.000
1162


184
VLRTQGPAAL
4.000
1163


241
RLHGDEERFF
4.000
1164


294
APWIMTPSTL
4.000
1165


142
GPSGYGGILL
4.000
1166


505
KCGHGRALLF
4.000
1167


17
AVCVAGVETL
4.000
1168


563
ECDIIDIPQL
4.000
1169


94
ISYHSSHEPL
4.000
1170


416
VVANGKEYPL
4.000
1171


491
ASPGACFKLF
3.600
1172


270
TLLDDSNEDF
3.600
1173


31
GSVPEGTEMF
3.600
1174


282
SPIFTDTVVF
3.000
1175


648
TNVCRKPFSF
3.000
1176


124
NCEGRQDRNF
3.000
1177


252
EGLSFPDAGF
3.000
1178


477
FVPAPDGKGF
3.000
1179


488
MLLASPGACF
3.000
1180


370
SPRNGELQDF
2.400
1181


468
VGHVDEFLSF
2.400
1182


359
APHKTLPVVF
2.400
1183


465
WLAVGHVDEF
2.200
1184


67
RWRFDATLEI
2.200
1185


597
LGIPKPFGPI
2.160
1186


288
TVVFRVAPWI
2.100
1187


646
CGTNVCRKPF
2.000
1188


310
VCRVRNNTCF
2.000
1189


258
DAGFTGLISF
2.000
1190


625
GLHCTFIDDF
2.000
1191


275
SNEDFSASPI
1.800
1192


375
ELQDFPYKRI
1.800
1193


43
YGTPGVDIYI
1.680
1194


343
RNDRWIQDEM
1.540
1195


339
QAENRNDRWI
1.500
1196


108
AVLYLTCVDI
1.500
1197


622
EPLGLHCTFI
1.500
1198


542
KFVQSCIDWN
1.500
1199


598
GIPKPFGPII
1.500
1200


529
NQVLSNKDLI
1.500
1201


590
MLVLGKHLGI
1.500
1202


318
CFVDAVAELA
1.260
1203


583
FFPDLVNMLV
1.260
1204


560
GLAECDIIDI
1.200
1205


517
VVDDEQVKTI
1.200
1206


140
VWGPSGYGGI
1.200
1207


69
RFDATLEIIV
1.200
1208


552
REVLKRELGL
1.200
1209


419
NGKEYPLGRI
1.200
1210


207
DAKRAQVFHI
1.000
1211


256
FPDAGFTGLI
1.000
1212


20
VAGVETLVDI
1.000
1213


41
EVYGTPGVDI
1.000
1214


653
KPFSFKWWNM
1.000
1215







v.2-A24-10mers: 184P1E2










3
IMTPSTLAPL
4.800
1216


7
STLAPLEVYV
0.180
1217


10
APLEVYVCRV
0.180
1218


8
TLAPLEVYVC
0.120
1219


5
TPSTLAPLEV
0.110
1220


9
LAPLEVYVCR
0.021
1221


4
MTPSTLAPLE
0.018
1222


1
PWIMTPSTLA
0.015
1223


2
WIMTPSTLAP
0.015
1224


6
PSTLAPLEVY
0.010
1225







v.3-A24-10mers: 184P1E2










10
VPDGKGFRML
4.800
1226


7
FVPVPDGKGF
3.000
1227


3
EFLSFVPVPD
0.105
1228


6
SFVPVPDGKG
0.099
1229


9
PVPDGKGFRM
0.090
1230


1
VDEFLSFVPV
0.018
1231


8
VPVPDGKGFR
0.018
1232


5
LSFVPVPDGK
0.014
1233


4
FLSFVPVPDG
0.010
1234


2
DEFLSFVPVP
0.001
1235



















TABLE XV





Pos
123456789
Score
SeqID















v.1-B7-9mers: 184P1E2










616
KVRSLLEPL
200.000
1236


326
LARKAGCKL
120.000
1237


142
GPSGYGGIL
80.000
1238


467
AVGHVDEFL
60.000
1239


3
LQRIVRVSL
60.000
1240


191
AALFDDHKL
36.000
1241


356
YVQAPHKTL
30.000
1242


256
FPDAGFTGL
24.000
1243


553
EVLKRELGL
20.000
1244


234
KVSYEVPRL
20.000
1245


480
APDGKGFRM
18.000
1246


417
VANGKEYPL
12.000
1247


579
KATAFFPDL
12.000
1248


491
ASPGACFKL
12.000
1249


359
APHKTLPVV
12.000
1250


312
RVRNNTCFV
10.000
1251


599
IPKPFGPII
8.000
1252


294
APWIMTPST
6.000
1253


442
VTQVVRDFL
6.000
1254


482
DGKGFRMLL
6.000
1255


79
VMNSPSNDL
6.000
1256


505
KCGHGRALL
6.000
1257


263
GLISFHVTL
4.000
1258


589
NMLVLGKHL
4.000
1259


246
EERFFVEGL
4.000
1260


402
SGLDSFGNL
4.000
1261


264
LISFHVTLL
4.000
1262


18
VCVAGVETL
4.000
1263


101
EPLPLAYAV
4.000
1264


529
NQVLSNKDL
4.000
1265


524
KTISINQVL
4.000
1266


97
HSSHEPLPL
4.000
1267


618
RSLLEPLGL
4.000
1268


111
YLTCVDISL
4.000
1269


434
LPGSSGRRV
4.000
1270


282
SPIFTDTVV
4.000
1271


298
MTPSTLPPL
4.000
1272


126
EGRQDRNFV
3.000
1273


72
ATLEIIVVM
3.000
1274


438
SGRRVTQVV
2.000
1275


13
HPTSAVCVA
2.000
1276


395
EPRDRSVSG
2.000
1277


604
GPIINGCCC
2.000
1278


530
QVLSNKDLI
2.000
1279


303
LPPLEVYVC
2.000
1280


239
VPRLHGDEE
2.000
1281


591
LVLGKHLGI
2.000
1282


289
VVFRVAPWI
2.000
1283


370
SPRNGELQD
2.000
1284


547
CIDWNREVL
1.800
1285


222
VCEAYRHVL
1.800
1286


64
DTRRWRFDA
1.500
1287


17
AVCVAGVET
1.500
1288


322
AVAELARKA
1.500
1289


21
AGVETLVDI
1.200
1290


166
NCDQHVHCL
1.200
1291


208
AKRAQVFHI
1.200
1292


584
FPDLVNMLV
1.200
1293


612
CLEEKVRSL
1.200
1294


31
GSVPEGTEM
1.000
1295


19
CVAGVETLV
1.000
1296


310
VCRVRNNTC
1.000
1297


221
DVCEAYRHV
1.000
1298


344
NDRWIQDEM
1.000
1299


184
VLRTQGPAA
1.000
1300


290
VFRVAPWIM
1.000
1301


172
HCLQDLEDM
1.000
1302


309
YVCRVRNNT
0.750
1303


107
YAVLYLTCV
0.600
1304


580
ATAFFPDLV
0.600
1305


451
HAQKVQPPV
0.600
1306


453
QKVQPPVEL
0.600
1307


598
GIPKPFGPI
0.600
1308


510
RALLFQGVV
0.600
1309


632
DDFTPYHML
0.600
1310


358
QAPHKTLPV
0.600
1311


192
ALFDDHKLV
0.600
1312


281
ASPIFTDTV
0.600
1313


71
DATLEIIVV
0.600
1314


458
PVELFVDWL
0.600
1315


6
IVRVSLEHP
0.500
1316


445
VVRDFLHAQ
0.500
1317


643
EVHCGTNVC
0.500
1318


183
MVLRTQGPA
0.500
1319


444
QVVRDFLHA
0.500
1320


392
VTREPRDRS
0.450
1321


143
PSGYGGILL
0.400
1322


377
QDFPYKRIL
0.400
1323


564
CDIIDIPQL
0.400
1324


492
SPGACFKLF
0.400
1325


368
FDSPRNGEL
0.400
1326


169
QHVHCLQDL
0.400
1327


622
EPLGLHCTF
0.400
1328


421
KEYPLGRIL
0.400
1329


346
RWIQDEMEL
0.400
1330


653
KPFSFKWWN
0.400
1331


504
QKCGHGRAL
0.400
1332


540
YNKFVQSCI
0.400
1333


104
PLAYAVLYL
0.400
1334


44
GTPGVDIYI
0.400
1335







v.2-B7-9mers: 184P1E2










3
MTPSTLAPL
4.000
1336


9
APLEVYVCR
0.600
1337


1
WIMTPSTLA
0.450
1338


8
LAPLEVYVC
0.300
1339


4
TPSTLAPLE
0.200
1340


7
TLAPLEVYV
0.200
1341


6
STLAPLEVY
0.030
1342


5
PSTLAPLEV
0.020
1343


2
IMTPSTLAP
0.010
1344







v.3-B7-9mers: 184P1E2










9
VPDGKGFRM
6.000
1345


7
VPVPDGKGF
0.400
1346


6
FVPVPDGKG
0.075
1347


1
DEFLSFVPV
0.020
1348


4
LSFVPVPDG
0.015
1349


3
FLSFVPVPD
0.010
1350


8
PVPDGKGFR
0.005
1351


2
EFLSFVPVP
0.001
1352


5
SFVPVPDGK
0.001
1353



















TABLE XVI





Pos
1234567890
Score
SeqID















v.1-B7-10mers: 184P1E2










395
EPRDRSVSGL
800.000
1354


294
APWIMTPSTL
240.000
1355


78
VVMNSPSNDL
90.000
1356


142
GPSGYGGILL
80.000
1357


604
GPIINGCCCL
80.000
1358


101
EPLPLAYAVL
80.000
1359


103
LPLAYAVLYL
80.000
1360


480
APDGKGFRML
72.000
1361


17
AVCVAGVETL
60.000
1362


425
LGRILIGGNL
40.000
1363


184
VLRTQGPAAL
40.000
1364


550
WNREVLKREL
40.000
1365


441
RVTQVVRDFL
30.000
1366


221
DVCEAYRHVL
30.000
1367


584
FPDLVNMLVL
24.000
1368


416
VVANGKEYPL
20.000
1369


653
KPFSFKWWNM
20.000
1370


452
AQKVQPPVEL
18.000
1371


588
VNMLVLGKHL
12.000
1372


466
LAVGHVDEFL
12.000
1373


192
ALFDDHKLVL
12.000
1374


490
LASPGACFKL
12.000
1375


622
EPLGLHCTFI
8.000
1376


457
PRVELFVDWL
8.000
1377


546
SCIDWNREVL
6.000
1378


114
CVDISLDCDL
6.000
1379


299
TPSTLPPLEV
6.000
1380


2
SLQRIVRVSL
6.000
1381


108
AVLYLTCVDI
6.000
1382


289
VVFRVAPWIM
5.000
1383


6
IVRVSLEHPT
5.000
1384


401
VSGLDSFGNL
4.000
1385


611
CCLEEKVRSL
4.000
1386


503
KQKCGHGRAL
4.000
1387


141
WGPSGYGGIL
4.000
1388


635
TPYHMLHGEV
4.000
1389


325
ELARKAGCKL
4.000
1390


317
TCFVDAVAEL
4.000
1391


263
GLISFHVTLL
4.000
1392


297
IMTPSTLPPL
4.000
1393


370
SPRNGELQDF
4.000
1394


65
TRRWRFDATL
4.000
1395


528
INQVLSNKDL
4.000
1396


165
DNCDQHVHCL
4.000
1397


168
DQHVHCLQDL
4.000
1398


94
ISYHSSHEPL
4.000
1399


262
TGLISFHVTL
4.000
1400


71
DATLEIIVVM
3.000
1401


434
LPGSSGRRVT
3.000
1402


326
LARKAGCKLT
3.000
1403


41
EVYGTPGVDI
3.000
1404


581
TAFFPDLVNM
3.000
1405


256
FPDAGFTGLI
2.400
1406


158
DPSCDVQDNC
2.000
1407


508
HGRALLFQGV
2.000
1408


288
TVVFRVAPWI
2.000
1409


392
VTREPRDRSV
2.000
1410


601
KPFGPIINGC
2.000
1411


239
VPRLHGDEER
2.000
1412


191
AALFDDHKLV
1.800
1413


612
CLEEKVRSLL
1.800
1414


47
GVDIYISPNM
1.500
1415


563
ECDIIDIPQL
1.200
1416


190
PAALFDDHKL
1.200
1417


176
DLEDMSVMVL
1.200
1418


376
LQDFPYKRIL
1.200
1419


33
VPEGTEMFEV
1.200
1420


359
APHKTLPVVF
1.200
1421


207
DAKRAQVFHI
1.200
1422


20
VAGVETLVDI
1.200
1423


582
AFFPDLVNML
1.200
1424


64
DTRRWRFDAT
1.000
1425


30
YGSVPEGTEM
1.000
1426


454
KVQPPVELFV
1.000
1427


162
DVQDNCDQHV
1.000
1428


522
QVKTISINQV
1.000
1429


504
QKCGHGRALL
0.600
1430


579
KATAFFPDLV
0.600
1431


355
GYVQAPHKTL
0.600
1432


597
LGIPKPFGPI
0.600
1433


419
NGKEYPLGRI
0.600
1434


281
ASPIFTDTVV
0.600
1435


375
ELQDFPYKRI
0.600
1436


358
QAPHKTLPVV
0.600
1437


280
SASPIFTDTV
0.600
1438


517
VVDDEQVKTI
0.600
1439


199
LVLHTSSYDA
0.500
1440


553
EVLKRELGLA
0.500
1441


8
RVSLEHPTSA
0.500
1442


183
MVLRTQGPAA
0.500
1443


445
VVRDFLHAQK
0.500
1444


312
RVRNNTCFVD
0.500
1445


616
KVRSLLEPLG
0.500
1446


309
YVCRVRNNTC
0.500
1447


516
GVVDDEQVKT
0.500
1448


43
YGTPGVDIYI
0.400
1449


578
KKATAFFPDL
0.400
1450


599
IPKPFGPIIN
0.400
1451


456
QPPVELFVDW
0.400
1452


233
DKVSYEVPRL
0.400
1453







v.2-B7-10mers: 184P1E2










10
APLEVYVCRV
12.000
1454


5
TPSTLAPLEV
4.000
1455


3
IMTPSTLAPL
4.000
1456


7
STLAPLEVYV
0.200
1457


8
TLAPLEVYVC
0.100
1458


9
LAPLEVYVCR
0.030
1459


2
WIMTPSTLAP
0.030
1460


4
MTPSTLAPLE
0.010
1461


6
PSTLAPLEVY
0.003
1462


1
PWIMTPSTLA
0.002
1463







v.3-B7-10mers: 184P1E2










10
VPDGKGFRML
24.000
1464


9
PVPDGKGFRM
0.500
1465


8
VPVPDGKGFR
0.200
1466


7
FVPVPDGKGF
0.100
1467


4
FLSFVPVPDG
0.015
1468


5
LSFVPVPDGK
0.010
1469


1
VDEFLSFVPV
0.006
1470


6
SFVPVPDGKG
0.002
1471


2
DEFLSFVPVP
0.001
1472


3
EFLSFVPVPD
0.001
1473



















TABLE XVII





Pos
123456789
Score
SeqID















v.1-B35-9 mers: 184P1E2










103
LPLAYAVLY
40.000
1474


478
VPAPDGKGF
30.000
1475


218
GPEDVCEAY
24.000
1476


599
IPKPFGPII
24.000
1477


492
SPGACFKLF
20.000
1478


622
EPLGLHCTF
20.000
1479


142
GPSGYGGIL
20.000
1480


31
GSVPEGTEM
15.000
1481


618
RSLLEPLGL
15.000
1482


532
LSNKDLINY
15.000
1483


480
APDGKGFRM
12.000
1484


399
RSVSGLDSF
10.000
1485


326
LARKAGCKL
9.000
1486


97
HSSHEPLPL
7.500
1487


579
KATAFFPDL
6.000
1488


256
FPDAGFTGL
6.000
1489


616
KVRSLLEPL
6.000
1490


491
ASPGACFKL
5.000
1491


191
AALFDDHKL
4.500
1492


101
EPLPLAYAV
4.000
1493


359
APHKTLPVV
4.000
1494


35
EGTEMFEVY
4.000
1495


434
LPGSSGRRV
4.000
1496


383
RILGPDFGY
4.000
1497


282
SPIFTDTVV
4.000
1498


653
KPFSFKWWN
4.000
1499


198
KLVLHTSSY
4.000
1500


72
ATLEIIVVM
4.000
1501


417
VANGKEYPL
3.000
1502


158
DPSCDVQDN
3.000
1503


3
LQRIVRVSL
3.000
1504


172
HCLQDLEDM
3.000
1505


241
RLHGDEERF
3.000
1506


482
DGKGFRMLL
3.000
1507


466
LAVGHVDEF
3.000
1508


229
VLGQDKVSY
3.000
1509


82
SPSNDLNDS
3.000
1510


303
LPPLEVYVC
3.000
1511


234
KVSYEVPRL
3.000
1512


45
TPGVDIYIS
3.000
1513


441
RVTQVVRDF
2.000
1514


402
SGLDSFGNL
2.000
1515


294
APWIMTPST
2.000
1516


604
GPIINGCCC
2.000
1517


524
KTISINQVL
2.000
1518


565
DIIDIPQLF
2.000
1519


454
KVQPPVELF
2.000
1520


457
PPVELFVDW
2.000
1521


505
KCGHGRALL
2.000
1522


32
SVPEGTEMF
2.000
1523


13
HPTSAVCVA
2.000
1524


186
RTQGPAALF
2.000
1525


43
YGTPGVDIY
2.000
1526


301
STLPPLEVY
2.000
1527


62
RADTRRWRF
1.800
1528


274
DSNEDFSAS
1.500
1529


131
RNFVDKRQW
1.500
1530


510
RALLFQGVV
1.200
1531


312
RVRNNTCFV
1.200
1532


540
YNKFVQSCI
1.200
1533


348
IQDEMELGY
1.200
1534


584
FPDLVNMLV
1.200
1535


395
EPRDRSVSG
1.200
1536


9
VSLEHPTSA
1.000
1537


356
YVQAPHKTL
1.000
1538


589
NMLVLGKHL
1.000
1539


79
VMNSPSNDL
1.000
1540


264
LISFHVTLL
1.000
1541


15
TSAVCVAGV
1.000
1542


467
AVGHVDEFL
1.000
1543


263
GLISFHVTL
1.000
1544


506
CGHGRALLF
1.000
1545


442
VTQVVRDFL
1.000
1546


98
SSHEPLPLA
1.000
1547


647
GTNVCRKPF
1.000
1548


553
EVLKRELGL
1.000
1549


253
GLSFPDAGF
1.000
1550


1
MSLQRIVRV
1.000
1551


117
ISLDCDLNC
1.000
1552


281
ASPIFTDTV
1.000
1553


18
VCVAGVETL
1.000
1554


655
FSFKWWNMV
1.000
1555


529
NQVLSNKDL
1.000
1556


649
NVCRKPFSF
1.000
1557


111
YLTCVDISL
1.000
1558


298
MTPSTLPPL
1.000
1559


259
AGFTGLISF
1.000
1560


437
SSGRRVTQV
1.000
1561


489
LLASPGACF
1.000
1562


126
EGRQDRNFV
0.900
1563


71
DATLEIIVV
0.900
1564


86
DLNDSHVQI
0.800
1565


21
AGVETLVDI
0.800
1566


254
LSFPDAGFT
0.750
1567


401
VSGLDSFGN
0.750
1568


203
TSSYDAKRA
0.750
1569


629
TFIDDFTPY
0.600
1570


344
NDRWIQDEM
0.600
1571


612
CLEEKVRSL
0.600
1572


22
GVETLVDIY
0.600
1573







v.2-B35-9 mers: 184P1E2










6
STLAPLEVY
2.000
1574


3
MTPSTLAPL
1.000
1575


8
LAPLEVYVC
0.450
1576


9
APLEVYVCR
0.400
1577


7
TLAPLEVYV
0.200
1578


4
TPSTLAPLE
0.200
1579


5
PSTLAPLEV
0.100
1580


1
WIMTPSTLA
0.100
1581


2
IMTPSTLAP
0.010
1582







v.3-B35-9 mers: 184P1E2










7
VPVPDGKGF
30.000
1583


9
VPDGKGFRM
12.000
1584


4
LSFVPVPDG
0.050
1585


1
DEFLSFVPV
0.020
1586


6
FVPVPDGKG
0.010
1587


3
FLSFVPVPD
0.010
1588


8
PVPDGKGFR
0.002
1589


2
EFLSFVPVP
0.001
1590


5
SFVPVPDGK
0.001
1591



















TABLE XVIII





Pos
1234567890
Score
SeqID















v.1-B35-10 mers: 184P1E2










395
EPRDRSVSGL
120.000
1592


370
SPRNGELQDF
90.000
1593


653
KPFSFKWWNM
80.000
1594


282
SPIFTDTVVF
30.000
1595


294
APWIMTPSTL
20.000
1596


98
SSHEPLPLAY
20.000
1597


103
LPLAYAVLYL
20.000
1598


359
APHKTLPVVF
20.000
1599


142
GPSGYGGILL
20.000
1600


101
EPLPLAYAVL
20.000
1601


604
GPIINGCCCL
20.000
1602


456
QPPVELFVDW
10.000
1603


581
TAFFPDLVNM
9.000
1604


372
RNGELQDFPY
8.000
1605


622
EPLGLHCTFI
8.000
1606


550
WNREVLKREL
6.000
1607


599
IPKPFGPIIN
6.000
1608


480
APDGKGFRML
6.000
1609


503
KQKCGHGRAL
6.000
1610


584
FPDLVNMLVL
6.000
1611


71
DATLEIIVVM
6.000
1612


94
ISYHSSHEPL
5.000
1613


31
GSVPEGTEMF
5.000
1614


401
VSGLDSFGNL
5.000
1615


491
ASPGACFKLF
5.000
1616


414
PPVVANGKEY
4.000
1617


21
AGVETLVDIY
4.000
1618


137
RQWVWGPSGY
4.000
1619


457
PPVELFVDWL
4.000
1620


347
WIQDEMELGY
4.000
1621


635
TPYHMLHGEV
4.000
1622


299
TPSTLPPLEV
4.000
1623


217
CGPEDVCEAY
4.000
1624


601
KPFGPIINGC
4.000
1625


207
DAKRAQVFHI
3.600
1626


531
VLSNKDLINY
3.000
1627


241
RLHGDEERFF
3.000
1628


364
LPVVFDSPRN
3.000
1629


490
LASPGACFKL
3.000
1630


184
VLRTQGPAAL
3.000
1631


258
DAGFTGLISF
3.000
1632


628
CTFIDDFTPY
3.000
1633


452
AQKVQPPVEL
3.000
1634


310
VCRVRNNTCF
3.000
1635


30
YGSVPEGTEM
3.000
1636


228
HVLGQDKVSY
3.000
1637


425
LGRILIGGNL
3.000
1638


466
LAVGHVDEFL
3.000
1639


256
FPDAGFTGLI
2.400
1640


419
NGKEYPLGRI
2.400
1641


204
SSYDAKRAQV
2.000
1642


192
ALFDDHKLVL
2.000
1643


158
DPSCDVQDNC
2.000
1644


289
VVFRVAPWIM
2.000
1645


546
SCIDWNREVL
2.000
1646


221
DVCEAYRHVL
2.000
1647


165
DNCDQHVHCL
2.000
1648


434
LPGSSGRRVT
2.000
1649


9
VSLEHPTSAV
2.000
1650


270
TLLDDSNEDF
2.000
1651


441
RVTQVVRDFL
2.000
1652


505
KCGHGRALLF
2.000
1653


611
CCLEEKVRSL
2.000
1654


33
VPEGTEMFEV
1.800
1655


477
FVPAPDGKGF
1.500
1656


650
VCRKPFSFKW
1.500
1657


468
VGHVDEFLSF
1.500
1658


343
RNDRWIQDEM
1.200
1659


392
VTREPRDRSV
1.200
1660


560
GLAECDIIDI
1.200
1661


630
FIDDFTPYHM
1.200
1662


579
KATAFFPDLV
1.200
1663


479
PAPDGKGFRM
1.200
1664


20
VAGVETLVDI
1.200
1665


648
TNVCRKPFSF
1.000
1666


300
PSTLPPLEVY
1.000
1667


528
INQVLSNKDL
1.000
1668


317
TCFVDAVAEL
1.000
1669


78
VVMNSPSNDL
1.000
1670


262
TGLISFHVTL
1.000
1671


281
ASPIFTDTVV
1.000
1672


263
GLISFHVTLL
1.000
1673


2
SLQRIVRVSL
1.000
1674


545
QSCIDWNREV
1.000
1675


297
IMTPSTLPPL
1.000
1676


436
GSSGRRVTQV
1.000
1677


437
SSGRRVTQVV
1.000
1678


17
AVCVAGVETL
1.000
1679


325
ELARKAGCKL
1.000
1680


646
CGTNVCRKPF
1.000
1681


625
GLHCTFIDDF
1.000
1682


141
WGPSGYGGIL
1.000
1683


168
DQHVHCLQDL
1.000
1684


588
VNMLVLGKHL
1.000
1685


252
EGLSFPDAGF
1.000
1686


465
WLAVGHVDEF
1.000
1687


416
VVANGKEYPL
1.000
1688


488
MLLASPGACF
1.000
1689


174
LQDLEDMSVM
0.900
1690


191
AALFDDHKLV
0.900
1691







v.2-B35-10 mers










10
APLEVYVCRV
8.000
1692


5
TPSTLAPLEV
4.000
1693


6
PSTLAPLEVY
1.000
1694


3
IMTPSTLAPL
1.000
1695


7
STLAPLEVYV
0.200
1696


8
TLAPLEVYVC
0.150
1697


9
LAPLEVYVCR
0.030
1698


4
MTPSTLAPLE
0.010
1699


2
WIMTPSTLAP
0.010
1700


1
PWIMTPSTLA
0.001
1701







v.3-B35-10 mers










10
VPDGKGFRML
6.000
1702


7
FVPVPDGKGF
1.500
1703


9
PVPDGKGFRM
0.400
1704


8
VPVPDGKGFR
0.200
1705


5
LSFVPVPDGK
0.050
1706


4
FLSFVPVPDG
0.010
1707


1
VDEFLSFVPV
0.006
1708


6
SFVPVPDGKG
0.001
1709


2
DEFLSFVPVP
0.001
1710


3
EFLSFVPVPD
0.001
1711
















TABLE XIX







Frequently Occurring Motifs











avrg. %




Name
identity
Description
Potential Function





zf-C2H2
34%
Zinc finger, C2H2 type
Nucleic acid-binding protein functions as





transcription factor, nuclear location





probable


cytochrome_b_N
68%
Cytochrome b(N-
membrane bound oxidase, generate




terminal)/b6/petB
superoxide


ig
19%
Immunoglobulin domain
domains are one hundred amino acids long





and include a conserved intradomain





disulfide bond.


WD40
18%
WD domain, G-beta repeat
tandem repeats of about 40 residues, each





containing a Trp-Asp motif. Function in





signal transduction and protein interaction


PDZ
23%
PDZ domain
may function in targeting signaling





molecules to sub-membranous sites


LRR
28%
Leucine Rich Repeat
short sequence motifs involved in protein-





protein interactions


pkinase
23%
Protein kinase domain
conserved catalytic core common to both





serine/threonine and tyrosine protein





kinases containing an ATP binding site and





a catalytic site


PH
16%
PH domain
pleckstrin homology involved in





intracellular signaling or as constituents of





the cytoskeleton


EGF
34%
EGF-like domain
30-40 amino-acid long found in the





extracellular domain of membrane-bound





proteins or in secreted proteins


rvt
49%
Reverse transcriptase





(RNA-dependent DNA





polymerase)



ank
25%
Ank repeat
Cytoplasmic protein, associates integral





membrane proteins to the cytoskeleton


oxidored_q1
32%
NADH-
membrane associated. Involved in proton




Ubiquinone/plastoquinone
translocation across the membrane




(complex I), various chains



efhand
24%
EF hand
calcium-binding domain, consists of a 12





residue loop flanked on both sides by a 12





residue alpha-helical domain


rvp
79%
Retroviral aspartyl protease
Aspartyl or acid proteases, centered on a





catalytic aspartyl residue


Collagen
42%
Collagen triple helix repeat
extracellular structural proteins involved in




(20 copies)
formation of connective tissue. The





sequence consists of the G-X-Y and the





polypeptide chains forms a triple helix.


fn3
20%
Fibronectin type III domain
Located in the extracellular ligand-binding





region of receptors and is about 200 amino





acid residues long with two pairs of





cysteines involved in disulfide bonds


7tm_1
19%
7 transmembrane receptor
seven hydrophobic transmembrane regions,




(rhodopsin family)
with the N-terminus located extracellularly





while the C-terminus is cytoplasmic.





Signal through G proteins
















TABLE XX





Motifs and Post-translational Modifications of 184P1E2







N-glycosylation site








88-91
NDSH


315-318
NNTC







cAMP- and cGMP-dependent protein kinase phosphorylation site








440-443
RRvT


578-581
KKaT







Protein kinase C phosphorylation site








65-67
TrR


370-372
SpR


438-440
SgR


533-535
SnK


575-577
TeR


656-658
SfK







Casein kinase II phosphorylation site








25-28
TlvD


32-35
SvpE


 98-101
SshE


113-116
TcvD


204-207
SsyD


255-258
SfpD


270-273
TllD


275-278
SneD


402-405
SglD


533-536
SnkD


546-549
SciD


619-622
SllE


629-632
TfiD







N-myristoylation site








331-336
GCklTI


432-437
GNlpGS


560-565
GLaeCD


625-630
GLhcTF


647-652
GTnvCR







Amidation site








438-441
sGRR
















TABLE XXI







Protein Properties of 184P1E2










Bioinformatic




Program
Outcome





184P1E2 v.1




ORF
ORF finder
bp42-2036 (includes stop)


Protein length

664 aa


Transmembrane
TM Pred
No TM


region
HMMTop
no TM, N terminus extracellular



Sosui
soluble protein



TMHMM
no TM, extracellular


Signal Peptide
Signal P
none


pI
pI/MW tool
pI5.3


Molecular weight
pI/MW tool
74.7 kDa


Localization
PSORT
Mitochondrial matrix space 59%,




mitochondrial inner membrane




29.9%, mitochondrial intermembrane




space 29.9%, mitochondrial outer




membrane 29.9%



PSORT II
47.8 cytoplasmic, 21.7% nuclear,




17.4% mitochondrial


Motifs
Pfam
Protein-arginine deiminase (PAD)



Prints
Cadherin signature, FAD dependent




pyridine nucleotide reductase




signature,



Blocks
G10 protein, urocanase,




phosphoglycerate kinase family,




developmental signaling protein




Wnt1family


184P1E2 v.2




ORF
ORF finder
bp42-2036 (includes stop)


Protein length

664 aa


Transmembrane
TM Pred
no TM


region
HMMTop
no TM, N terminus extracellular



Sosui
soluble protein



TMHMM
no TM, extracellular


Signal Peptide
Signal P
none


pI
pI/MW tool
pI5.3


Molecular
pI/MW tool
74.7 kDa


weight




Localization
PSORT
Mitochondrial matrix space 59%,




mitochondrial inner membrane




29.9%, mitochondrial intermembrane




space 29.9%, mitochondrial outer




membrane 29.9%



PSORT II
47.8% cytoplasmic, 21.7% nuclear,




17.4% mitochondrial


Motifs
Pfam
Protein-arginine deiminase (PAD)



Prints
Cadherin signature, FAD dependent




pyridine nucleotide reductase




signature



Blocks
G10 protein, urocanase,




phosphoglycerate kinase family,




developmental signaling protein




Wnt1family


184P1E2 v.3




ORF
ORF finder
bp42-2036 (includes stop)


Protein length

664 aa


Transmembrane
TM Pred
no TM


region
HMMTop
no TM, N terminus extracellular



Sosui
soluble protein



TMHMM
no TM, extracellular


Signal Peptide
Signal P
none


pI
pI/MW tool
pI5.3


Molecular
pI/MW tool
74.7 kDa


weight




Localization
PSORT
Mitochondrial matrix space 59%,




mitochondrial inner membrane




29.9%, mitochondrial intermembrane




space 29.9%, mitochondrial outer




membrane 29.9%



PSORT II
47.8 cytoplasmic, 21.7% nuclear,




17.4% mitochondrial


Motifs
Pfam
Protein-arginine deiminase (PAD)



Prints
Cadherin signature, FAD dependent




pyridine nucleotide reductase




signature



Blocks
G10 protein, urocanase,




phosphoglycerate kinase family,




developmental signaling protein




Wnt1family
















TABLE XXII







184P1E2 v.1: HLA Peptide Scoring


Results A1 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






99
S H E P L P L A Y
37
1712



348
I Q D E M E L C Y
31
1713



301
S T L P P L E V Y
29
1714



218
C P E D V C E A Y
27
1715



373
N G E L Q D F P Y
27
1716



22
G V E T L V D I Y
26
1717



43
Y G T P G V D I Y
24
1718



532
L S N K D L I N Y
23
1719



620
L L E P L G L H C
22
1720



88
N D S H V Q I S Y
21
1721



236
S Y E V P R L H G
20
1722



285
F T D T V V F R V
20
1723



35
E C T F N F F V Y
19
1724



103
L P L A Y A V L Y
18
1725



629
T F I D D F T P Y
18
1726



138
Q W V W G P S G Y
17
1727



193
L F D D H K L V L
17
1728



229
V L G Q D K V S Y
17
1729



383
R I L G P D F G Y
17
1730



566
I I D I P Q L F K
17
1731



36
G T E M F E V Y G
16
1732



198
K L V L H T S S Y
16
1733



256
F P D A C F T G L
16
1734



415
P V V A N G K E Y
16
1735



574
K T E R K K A T A
16
1736



584
F P D L V N M L V
16
1737



156
R D D P S C D V Q
15
1738



319
F V D A V A F L A
15
1739



613
L F E K V R S L L
15
1740



10
S L E H P T S A V
14
1741



160
S C D V Q D N C D
14
1742



222
V C E A Y R H V L
14
1743



244
G D E F R F F V E
14
1744



265
I S F H V T L L D
14
1745



393
T F E P R D R S V
14
1746



403
G L D S F C N L E
14
1747



518
V D D E Q V K T I
14
1748



561
L A E C D I I D I
14
1749



563
E C D I I D I P Q
14
1750



26
L V D I Y G S V P
13
1751



44
G T P C V D I Y I
13
1752



55
N N E R C R E R A
13
1753



73
T L E I I V V N N
13
1754



118
S L D C D L N C E
13
1755



128
R Q D R N F V D K
13
1756



143
P S G Y C G I L L
13
1757



144
S G Y G G I L L V
13
1758



194
F D D H K L V L H
13
1759



205
S Y D A K R A Q V
13
1760



245
D F E R F F V F G
13
1761



305
P L E V Y V C R V
13
1762



362
K T L P V V F D S
13
1763



367
V F D S P R N G E
13
1764



455
V Q P P V E L F V
13
1765



480
A P D G K G F R H
13
1766



500
F Q E K Q K C G H
13
1767



517
V V D D E Q V K T
13
1768



526
I S I N Q V L S N
13
1769



634
F T P Y H M L H G
13
1770



33
V P E C T E M F E
12
1771



39
M F E V Y G T P G
12
1772



84
S N D L N D S H V
12
1773



133
F V D K R Q W V W
12
1774



176
D L E D M S V M V
12
1775



177
L F D M S V M V L
12
1776



231
G Q D K V S Y E V
12
1777



250
F V E C L S F P D
12
1778



271
L L D D S N E D F
12
1779



275
S N E D F S A S P
12
1780



323
V A E L A R K A G
12
1781



351
E M E L G Y V Q A
12
1782



386
G P D F C Y V T R
12
1783



409
N L E V S P P V V
12
1784



412
V S P P V V A N G
12
1785



471
V D E F L S F V P
12
1786



519
D D E Q V K T I S
12
1787



547
C I D W N R F V L
12
1788



556
K R E L G L A F C
12
1789



612
C L E E K V R S L
12
1790



47
C V D I Y I S P N
11
1791



62
R A D T R R W R F
11
1792



87
L N D S H V Q I S
11
1793



124
N C E G R Q D F N
11
1794



155
D R D D P S C D V
11
1795



163
V Q D N C D Q H V
11
1796



166
N C D Q H V H C L
11
1797



243
H G D E E R F F V
11
1798



300
P S T L P P L E V
11
1799



339
Q A E N R N D R W
11
1800



376
L Q D F P Y K R I
11
1801



396
P R D R S V S G L
11
1802



420
G K E Y P L G R I
11
1803



428
I L I G G N L P G
11
1804



446
V R D F L H A Q K
11
1805



458
P V E L F V D W L
11
1806



462
F V D W L A V G H
11
1807



534
N K D L I N Y N K
11
1808



630
F I D D F T P Y H
11
1809



59
G R E R A D T R R
10
1810



69
R F D A T L E I I
10
1811



81
N S P S N D L N D
10
1812



97
H S S H E P L P L
10
1813



114
C V D I S L D C D
10
1814



117
I S L D C D L N C
10
1815



120
D C D L N C E G R
10
1816



153
N C D R D D P S C
10
1817



174
L Q D L E D M S V
10
1818



186
R T Q G P A A L F
10
1819



187
T Q G P A A L F D
10
1820



219
P E D V C E A Y R
10
1821



272
L D D S N E D F S
10
1822



276
N E D F S A S P I
10
1823



297
I M T P S T L P P
10
1824



343
R N D R W I Q D E
10
1825



349
Q D E M E L G Y V
10
1826



370
S P R N G E L Q D
10
1827



442
V T Q V V R D F L
10
1828



470
H V D E F L S F V
10
1829



551
N R E V L K R E L
10
1830



618
R S L L E P L G L
10
1831



631
I D D F T P Y H M
10
1832



641
H C E V H C C T N
10
1833



31
C S V P E C T E M
9
1834



98
S S H E P L P L A
9
1835



111
Y L T C V D I S L
9
1836



171
V H C L Q D L E D
9
1837



202
H T S S Y D A K R
9
1838



235
V S Y E V P R L H
9
1839



280
S A S P I F T D T
9
1840



316
N T C F V D A V A
9
1841



392
V T R E P R D R S
9
1842



404
L D S F C N L E V
9
1843



422
E Y P L C R I L I
9
1844



423
Y P L C R I L I G
9
1845



444
Q V V R D F L H A
9
1846



469
G H V D E F L S F
9
1847



506
C G H G R A L L F
9
1848



549
D W N R E V L K R
9
1849



580
A T A F F P D L V
9
1850



5
R I V R V S L E H
8
1851



1
P T S A V C V A G
8
1852



20
V A G V E T L V D
8
1853



72
A T L E I I V V M
8
1854



10
P L A Y A V L Y L
8
1855



112
L T C V D I S L D
8
1856



145
G Y G G I L L V N
8
1857



254
L S F P D A G F T
8
1858



266
S F H V T L L D D
8
1859



38
I L G P D F G Y V
8
1860



45
K V Q P P V E L F
8
1861



483
G K G F R M L L A
8
1862



491
A S P G A C F K L
8
1863



524
K T I S I N Q V L
8
1864



531
V L S N K D L I N
8
1865



567
I D I P Q L F K T
8
1866



581
T A F F P D L V N
8
1867



614
E E K V R S L L E
8
1868



625
C L H C T F I D D
8
1869



23
V E T L V D I Y G
7
1870



64
D T R R W R F D A
7
1871



80
M N S P S N D L N
7
1872



106
A Y A V L Y L T C
7
1873



179
D M S V M V L R T
7
1874



195
D D H K L V L H T
7
1875



224
F A Y R H V L G Q
7
1876



248
R F F V E G L S F
7
1877



258
D A G F T C L I S
7
1878



259
A C F T C L I S F
7
1879



261
F T G L I S F H V
7
1880



269
V T L L D D S N E
7
1881



274
D S N E D F S A S
7
1882



287
D T V V F R V A P
7
1883



291
F R V A P W I M T
7
1884



320
V D A V A E L A R
7
1885



328
R K A C C K L T I
7
1886



334
L T I C P Q A E N
7
1887



357
V Q A P H K T L P
7
1888



369
D S P R N G E L Q
7
1889



378
D F P Y K R I L G
7
1890



379
F P Y K R I L G P
7
1891



398
D R S V S G L D S
7
1892



419
N C K E Y P L C R
7
1893



437
S S G R R V T Q V
7
1894



460
E L F V D W L A V
7
1895



492
S P G A C F K L F
7
1896



555
L K R E L G L A E
7
1897



585
P D L V N M L V L
7
1898



587
L V N M L V L G K
7
1899



591
L V L G K H L G I
7
1900



633
D F T P Y H M L H
7
1901



647
G T N V C R K P F
7
1902



24
E T L V D T Y G S
6
1903



51
Y I S P N M E R G
6
1904



68
W R F D A T L S I
6
1905



89
D S H V Q I S Y H
6
1906



113
T C V D I S L D C
6
1907



140
V W G P S G Y C G
6
1908



159
P S C D V Q D N C
6
1909



168
D Q H V H C L Q D
6
1910



180
M S V M V L R T Q
6
1911



204
S S Y D A K R A Q
6
1912



237
Y E V P R L H G D
6
1913



279
F S A S P I F T D
6
1914



286
T D T V V F R V A
6
1915



298
M T P S T L P P L
6
1916



335
T I C P Q A S N R
6
1917



358
Q A P H K T L P V
6
1918



363
T L P V V F D S P
6
1919



368
F D S P R N C S L
6
1920



399
R S V S G L D S F
6
1921



405
D S F G N L E V S
6
1922



411
E V S P P V V A N
6
1923



432
G N L P G S S C R
6
1924



436
G S S C R R V T Q
6
1925



476
S F V P A P D C K
6
1926



478
V P A P D G K G F
6
1927



481
P D G K G F R M L
6
1928



484
K C F R M L L A S
6
1929



490
L A S P C A C F K
6
1930



493
P G A C F K L F Q
6
1931



507
G H G R A L L F Q
6
1932



511
A L L F Q G V V D
6
1933



553
S V L K R E L G L
6
1934



600
P K P F G P I I N
6
1935



601
K P F G P I I N G
6
1936



607
I N G C C C L E E
6
1937



615
E K V R S L L S P
6
1938



619
S L L E P L C L H
6
1939



628
C T F I D D F T P
6
1940



651
C R K P F S F K W
6
1941



655
F S F K W W N M V
6
1942



2
S L Q R I V R V S
5
1943



9
V S L E H P T S A
5
1944



11
L E H P T S A V C
5
1945



15
T S A V C V A G V
5
1946



16
S A V C V A G V E
5
1947



19
C V A G V E T L V
5
1948



32
S V P E G T E M F
5
1949



70
F D A T L S I I V
5
1950



71
D A T L S I I V V
5
1951



83
P S N D L N D S H
5
1952



184
V L R T Q G P A A
5
1953



209
K R A Q V F H I C
5
1954



281
A S P I F T D T V
5
1955



302
T L P P L E V Y V
5
1956



309
Y V C R V R N N T
5
1957



327
A R K A G C K L T
5
1958



330
A G C K L T I C P
5
1959



377
Q D F P Y K R I L
5
1960



401
V S G L D S F G N
5
1961



402
S G L D S F C N L
5
1962



421
K S Y P L G R I L
5
1963



424
P L G R I L I C G
5
1964



427
R I L I G G N L P
5
1965



438
S G R R V T Q V V
5
1966



459
V E L F V D W L A
5
1967



475
L S F V P A P D G
5
1968



505
K C C H G R A L L
5
1969



560
G L A E C D I I D
5
1970



582
A F F P D L V N M
5
1971



588
V N M L V L G K H
5
1972



590
M L V L C K H L G
5
1973



593
L G K H L G I P K
5
1974



599
I P K P F G P I I
5
1975



650
V C R K P F S F K
5
1976



1
M S L Q R I V R V
4
1977



18
V C V A G V E T L
4
1978



46
P C V D I Y I S P
4
1979



52
I S P N M E R G R
4
1980



94
I S Y H S S H E P
4
1981



96
Y H S S H E P L P
4
1982



101
F P L P L A Y A V
4
1983



105
L A Y A V L Y L T
4
1984



167
C D Q H V H C L Q
4
1985



178
E D M S V M V L R
4
1986



192
A L F D D H K L V
4
1987



203
T S S Y D A K R A
4
1988



208
A K R A Q V F H I
4
1989



210
R A Q V F H I C G
4
1990



216
I C G P E D V C E
4
1991



227
R H V L G Q D K V
4
1992



228
H V L G Q D K V S
4
1993



247
E R F F V E G L S
4
1994



251
V E G L S F P D A
4
1995



262
T G L I S F H V T
4
1996



278
D F S A S P I F T
4
1997



292
R V A P W I M T P
4
1998



342
N R N D R W I Q D
4
1999



397
R D R S V S C L D
4
2000



413
S P P V V A N G K
4
2001



435
P G S S G R R V T
4
2002



449
F L H A Q K V Q P
4
2003



452
A Q K V Q P P V E
4
2004



464
D W L A V G H V D
4
2005



474
F L S F V P A P D
4
2006



477
F V P A P D G K G
4
2007



482
D G K C F R M L L
4
2008



495
A C F K L F Q E K
4
2009



512
L L F Q G V V D D
4
2010



513
L F Q G V V D D E
4
2011



520
D E Q V K T I S I
4
2012



522
Q V K T I S I N Q
4
2013



527
S I N Q V L S N K
4
2014



540
Y N K F V Q S C I
4
2015



542
K F V Q S C I D W
4
2016



545
Q S C I D W N R E
4
2017



554
V L K R S L G L A
4
2018



594
C K H L G I P K P
4
2019



597
L C I P K P F C P
4
2020



598
G I P K P F C P I
4
2021



602
P F G P I I N C C
4
2022



637
Y H M L H C S V H
4
2023



646
C C T N V C R K P
4
2024



656
S F K W W N M V P
4
2025



3
L Q R I V R V S L
3
2026



4
Q R I V R V S L E
3
2027



27
V D I Y G S V P E
3
2028



30
Y C S V P E C T E
3
2029



40
F E V Y G T P C V
3
2030



42
V Y G T P C V D I
3
2031



53
S P N M E R G R E
3
2032



65
T R R W R F D A T
3
2033



74
L E I I V V M N S
3
2034



107
Y A V L Y L T C V
3
2035



123
L N C E G R Q D R
3
2036



129
Q D R N F V D K R
3
2037



134
V D K R Q W V W G
3
2038



142
G P S G Y G G I L
3
2039



148
G I L L V N C D R
3
2040



181
S V M V L R T Q G
3
2041



189
G P A A L F D D H
3
2042



191
A A L F D D H K L
3
2043



206
Y D A K R A Q V F
3
2044



214
F H I C G P E D V
3
2045



215
H I C G P E D V C
3
2046



225
A Y R H V L G Q D
3
2047



226
Y R H V L G Q D K
3
2048



249
F F V E G L S F P
3
2049



255
S F P D A G F T G
3
2050



257
P D A G F T G L I
3
2051



264
L I S F H V T L L
3
2052



282
S P I F T D T V V
3
2053



289
V V F R V A P W I
3
2054



296
W I M T P S T L P
3
2055



306
L E V Y V C R C R
3
2056



314
R N N T C F V D A
3
2057



322
A V A E L A R K A
3
2058



326
L A R K A G C K L
3
2059



345
D R W I Q D E M E
3
2060



352
M E L G Y V Q A P
3
2061



353
E L G Y V Q A P H
3
2062



356
Y V Q A P H K T L
3
2063



374
G E L Q D F P Y K
3
2064



375
E L Q D F P Y K R
3
2065



381
Y K R I L G P D F
3
2066



387
P D F G Y N T R E
3
2067



400
S V S G L D S F G
3
2068



414
P P V V A M G K E
3
2069



416
V V A N G K E Y P
3
2070



418
A N G K E Y P L G
3
2071



440
R R V T Q V V R D
3
2072



441
R V T Q V V R D F
3
2073



457
P P V E L F V D M
3
2074



465
W L A V G H V D E
3
2075



467
A V G H V D E F L
3
2076



468
V G H V D E F L S
3
2077



472
D E F L S F V P A
3
2078



489
L L A S P G A C F
3
2079



497
F K L F Q E K Q K
3
2080



498
K L F Q E K Q K C
3
2081



509
G R A L L F Q C V
3
2082



514
F Q G V V D D E Q
3
2083



536
D L I N Y N K F V
3
2084



538
I N Y N K F V Q S
3
2085



546
S C I D W N R E V
3
2086



552
R E V L K R E L G
3
2087



568
D I P Q L F K T E
3
2088



571
Q L F K T E R K K
3
2089



572
L F K T E R K K A
3
2090



578
K K A T A F F P D
3
2091



579
K A T A F F P D L
3
2092



583
F F P D L V N M L
3
2093



586
D L V N M L V L G
3
2094



606
I I N G C C C L E
3
2095



617
V R S L L E P L G
3
2096



639
M L H G E V H C G
3
2097



643
E V H C G T N V C
3
2098



644
V H C G T N V C R
3
2099



652
R K P F S F K W W
3
2100



13
H P T S A V C V A
2
2101



17
A V C V A G V E T
2
2102



28
D I Y G S V P E G
2
2103



29
I Y G S V P E G T
2
2104



38
E M F E V Y G T P
2
2105



41
E V Y C T P G V D
2
2106



50
I Y I S P N M F R
2
2107



67
R W R F D A T L E
2
2108



75
F I I V V M N S P
2
2109



79
V M N S P S N D L
2
2110



82
S P S N D L N D S
2
2111



90
S H V Q I S Y H S
2
2112



92
V Q I S Y H S S H
2
2113



95
S Y H S S H E P L
2
2114



102
P L P L A Y A V L
2
2115



108
A V L Y L T C V D
2
2116



109
V L Y L T C V D I
2
2117



116
D I S L D C D L N
2
2118



122
D L N C E G R Q D
2
2119



125
C E G R Q D R N F
2
2120



127
G R Q D R N F V D
2
2121



130
D R N F V D K R Q
2
2122



136
K R Q W V W C P S
2
2123



139
W V W G P S G Y G
2
2124



141
W C P S G Y C G I
2
2125



146
Y G G I L L V N C
2
2126



165
D N C D Q H V H C
2
2127



170
H V H C L Q D L E
2
2128



173
C L Q D L E D M S
2
2129



185
L R T Q G P A A L
2
2130



196
D H K L V L H T S
2
2131



200
V L H T S S Y D A
2
2132



213
V F H I C G P E D
2
2133



223
C E A Y R H V L G
2
2134



239
V P R L H G D E E
2
2135



240
P R L H C D E E R
2
2136



241
R L H G D E E R F
2
2137



253
G L S F P D A G F
2
2138



299
T P S T L P P L E
2
2139



303
L P P L E V Y V C
2
2140



304
P P L F V Y V C R
2
2141



308
V Y V C R V R N N
2
2142



310
V C R V R N N T C
2
2143



313
V R N N T C F V D
2
2144



347
W I Q D E M E L G
2
2145



350
D E M E L G Y V Q
2
2146



354
L G Y V Q A P H K
2
2147



355
G Y V Q A P H K T
2
2148



359
A P H K T L P V V
2
2149



361
H K T L P V V F D
2
2150



382
K R I L G P D F G
2
2151



385
L G P D F G Y V T
2
2152



388
D F G Y V T R E P
2
2153



389
F G Y V T R E P R
2
2154



391
Y V T R E P R D R
2
2155



394
R E P R D R S V S
2
2156



406
S F G N L E V S P
2
2157



407
F G N L E V S P P
2
2158



408
G N L E V S P P V
2
2159



410
L F V S P P v V A
2
2160



417
V A N G K E Y P L
2
2161



431
G G N L P G S S G
2
2162



433
N L P G S S G R R
2
2163



443
T Q V V R D F L H
2
2164



447
R D F L H A Q K V
2
2165



451
H A Q K V Q P P V
2
2166



463
V D W L A V G H V
2
2167



466
L A V G H V D E F
2
2168



479
P A P D G K G F R
2
2169



496
C F K L F Q E K Q
2
2170



503
K Q K C G H G R A
2
2171



504
Q K C G H G R A L
2
2172



525
T I S I N Q V L S
2
2173



528
I N Q V L S N K D
2
2174



530
Q V L S N K D L I
2
2175



533
S N K D L I N Y N
2
2176



535
K D L I N Y N K F
2
2177



541
N K F V Q S C I D
2
2178



548
I D W N R E V L K
2
2179



550
W N R E V L K R E
2
2180



565
D I I D I P Q L F
2
2181



592
V L G K H L G I P
2
2182



596
H L G I P K P F G
2
2183



603
F G P I I N C C C
2
2184



605
P I I N C C C C L
2
2185



609
G C C C L E E K V
2
2186



621
L E P L G L H C T
2
2187



632
D D F T P Y H M L
2
2188



638
H M L H G E V H C
2
2189



640
L H G E V H C G T
2
2190



7
V R V S L E H P T
1
2191



21
A C V E T L V D I
1
2192



25
T L V D I Y G S V
1
2193



37
T E M F E V Y C T
1
2194



48
V D I Y I S P N M
1
2195



49
D I Y I S P N M E
1
2196



54
P N M E R G R E R
1
2197



56
M E R G R E R A D
1
2198



57
E R G R E R A D T
1
2199



61
E R A D T R R W R
1
2200



63
A D T R R W R F D
1
2201



76
I I V V M N S P S
1
2202



78
V V M N S P S N D
1
2203



85
N D L N D S H V Q
1
2204



86
D L N D S H V Q I
1
2205



100
H E P L P L A Y A
1
2206



110
L Y L T C V D I S
1
2207



115
V D I S L D C D L
1
2208



119
L D C D L N C E G
1
2209



126
E G R Q D R N F V
1
2210



132
N F V D K R Q W V
1
2211



135
D K R Q W V W G P
1
2212



137
R Q W V W G P S G
1
2213



147
C C I L L V N C D
1
2214



149
I L L V N C D R D
1
2215



150
L L V N C D R D D
1
2216



151
L V N C D R D D P
1
2217



152
V N C D R D D P S
1
2218



158
D P S C D V Q D N
1
2219



161
C D V Q D N C D Q
1
2220



164
Q D N C D Q H V H
1
2221



175
Q D L E D M S V M
1
2222



182
V M V L R T Q G P
1
2223



211
A Q V F H I C G P
1
2224



217
C G P E D V C E A
1
2225



230
L G Q D K V S Y E
1
2226



232
Q D K V S Y E V P
1
2227



233
D K V S Y E V P R
1
2228



234
K V S Y E V P R L
1
2229



246
E E R F F V E C L
1
2230



260
C F T G L I S F H
1
2231



263
C L I S F H V T L
1
2232



267
F H V T L L D D S
1
2233



270
T L L D D S N E D
1
2234



283
P I F T D T V V F
1
2235



284
I F T D T V V F R
1
2236



288
T V V F R V A P W
1
2237



290
V F R V A P W I M
1
2238



293
V A P W I M T P S
1
2239



294
A P W I M T P S T
1
2240



307
E V Y V C R V R N
1
2241



311
C R V R N N T C F
1
2242



315
N N T C F V D A V
1
2243



318
C F V D A V A E L
1
2244



324
A E L A R K A G C
1
2245



325
E L A R K A C C K
1
2246



333
K L T I C P Q A E
1
2247



337
C P Q A E N R N D
1
2248



338
P Q A E N R N D R
1
2249



340
A E N R N D R W I
1
2250



341
E N R N D R W I Q
1
2251



360
P H K T L P V V F
1
2252



366
V V F D S P R N G
1
2253



371
P R N G E L Q D F
1
2254



426
C R I L I C G N L
1
2255



429
L I G G N L P G S
1
2256



434
L P G S S G R R V
1
2257



439
G R R V T Q V V R
1
2258



445
V V R D F L H A Q
1
2259



448
D F L H A Q K V Q
1
2260



450
L H A Q K V Q P P
1
2261



453
Q K V Q P P V E L
1
2262



456
Q P P V E L F V D
1
2263



461
L F V D W L A V G
1
2264



473
E F L S F V P A P
1
2265



486
F R M L L A S P G
1
2266



488
M L L A S P G A C
1
2267



502
E K Q K C G H G R
1
2268



510
R A L L F Q G V V
1
2269



516
G V V D D E Q V K
1
2270



523
V K T I S I N Q V
1
2271



529
N Q V L S N K D L
1
2272



537
L I N Y N K F V Q
1
2273



539
N Y N K F V Q S C
1
2274



543
F V Q S C I D W N
1
2275



544
V Q S C I D W N R
1
2276



557
R E L G L A E C D
1
2277



558
E L G L A E C D I
1
2278



562
A E C D I I D I P
1
2279



573
F K T E R K K A T
1
2280



589
N M L V L G K H L
1
2281



595
K H L G I P K P F
1
2282



604
G P I I N G C C C
1
2283



610
C C C L E E K V R
1
2284



622
E P L G L H C T F
1
2285



623
P L G L H C T F I
1
2286



642
G E V H C G T N V
1
2287



645
H C G T N V C R K
1
2288










184P1E2 v.2: HLA Peptide Scoring


Results A1 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






6
S T L A P L E V Y
25
2289



2
I M T P S T L A P
10
2290



5
P S T L A P L E V
10
2291



3
M T P S T L A P L
6
2292



7
T L A P L E V Y V
5
2293



1
W I M T P S T L A
3
2294



9
A P L E V Y V C R
3
2295



4
T P S T L A P L E
2
2296



8
L A P L E V Y V C
2
2297










184P1E2 v.3: HLA Peptide Scoring


Results A1 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






9
V P D G K G F R M
13
2298



5
S F V P V P D G K
8
2299



4
L S F V P V P D G
6
2300



7
V P V P D G K G F
6
2301



3
F L S F V P V P D
4
2302



6
F V P V P D G K G
4
2303



1
D E F L S F V P V
3
2304



2
E F L S F V P V P
2
2305



8
P V P D G K G F R
2
2306
















TABLE XXIII







184P1E2 v.1: HLA Peptide Scoring


Results A0201 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






263
G L I S F H V T L
28
2307



612
C L E E K V R S L
28
2308



10
S L E H P T S A V
25
2309



302
T L P P L E V Y V
25
2310



384
I L G P D F G Y V
24
2311



25
T L V D I Y G S V
23
2312



104
P L A Y A V L Y L
23
2313



111
Y L T C V D I S L
23
2314



192
A L F D D H K L V
23
2315



264
L I S F H V T L L
23
2316



79
V M N S P S N D L
22
2317



109
V L Y L T C V D I
22
2318



144
S G Y C C I L L V
22
2319



86
D L N D S H V Q I
21
2320



107
Y A V L Y L T C V
21
2321



176
D L E D M S V M V
21
2322



305
P L E V Y V C R V
21
2323



470
H V D E F L S F V
21
2324



512
L L F Q C V V D D
21
2325



536
D L I N Y N K F V
21
2326



591
L V L C K H L C I
21
2327



619
S L L E P L C L H
21
2328



72
A T L E I I V V M
20
2329



191
A A L F D D H K L
20
2330



409
N L E V S P P V V
20
2331



417
V A N C K E Y P L
20
2332



460
E L F V D W L A V
20
2333



589
N M L V L C K H L
20
2334



616
K V R S L L E P L
20
2335



15
T S A V C V A C V
19
2336



18
V C V A C V E T L
19
2337



102
P L P L A Y A V L
19
2338



234
K V S Y E V P R L
19
2339



285
F T D T V V F R V
19
2340



298
M T P S T L P P L
19
2341



318
C F V D A V A E L
19
2342



428
I L I G G N L P G
19
2343



605
P I I N G C C C L
19
2344



1
M S L Q R I V R V
18
2345



19
C V A G V E T L V
18
2346



21
A G V F T L V D I
18
2347



326
L A R K A G C K L
18
2348



356
Y V Q A P H K T L
18
2349



358
Q A P H K T L P V
18
2350



429
L I G C N L P G S
18
2351



54
C I D W N R E V L
18
2352



55
V L K R E L G L A
18
2353



580
A T A F F P D L V
18
2354



583
F F P D L V N M L
18
2355



598
G I P K P F G P I
18
2356



28
D I Y G S V P E G
17
2357



118
S L D C D L N C E
17
2358



184
V L R T Q G P A A
17
2359



281
A S P I F T D T V
17
2360



322
A V A E L A R K A
17
2361



359
A P H K T L P V V
17
2362



489
L L A S P C A C F
17
2363



510
R A L L F Q C V V
17
2364



524
K T I S I N Q V L
17
2365



546
S C I D W N R B V
17
2366



561
L A E C D I I D I
17
2367



564
C D I I D I P Q L
17
2368



639
M L H C E V H C G
17
2369



2
S L Q R I V R V S
16
2370



3
L Q R I V R V S L
16
2371



40
F E V Y C T P G V
16
2372



66
R R W R F D A T L
16
2373



71
D A T L S I I V V
16
2374



105
L A Y A V L Y L T
16
2375



177
L E D M S V M V L
16
2376



179
D M S V M V L R T
16
2377



185
L R T Q G P A A L
16
2378



200
V L H T S S Y D A
16
2379



270
T L L D D S N E D
16
2380



312
R V R N N T C F V
16
2381



404
L D S F G N B E V
16
2382



408
C N B E V S P P V
16
2383



437
S S G R R V T Q V
16
2384



463
V D W L A V C H V
16
2385



465
W L A V G H V D E
16
2386



467
A V G H V D E F L
16
2387



487
R M L L A S P G A
16
2388



491
A S P G A C F K L
16
2389



518
V D D H Q V K T I
16
2390



567
I D I P Q L F K T
16
2391



579
K A T A F F P D L
16
2392



44
G T P G V D I Y I
15
2393



68
W R F D A T L E I
15
2394



73
T L E I I V V M N
15
2395



166
N C D Q H V H C L
15
2396



198
K L V L H T S S Y
15
2397



205
S Y D A K R A Q V
15
2398



214
F H I C G P E D V
15
2399



221
D V C E A Y R H V
15
2400



229
V L G Q D K V S Y
15
2401



261
F T C L I S F H V
15
2402



289
V V F R V A P W I
15
2403



301
S T L P P L E V Y
15
2404



315
N N T C F V D A V
15
2405



328
R K A G C K L T I
15
2406



368
F D S P R N G E L
15
2407



402
S G L D S F G N L
15
2408



421
K E Y P L C R I L
15
2409



442
V T Q V V R D F L
15
2410



451
H A Q K V Q P P V
15
2411



453
Q K V Q P P V E L
15
2412



458
P V E L F V D W L
15
2413



505
K C G H G R A L L
15
2414



509
G R A L L F Q G V
15
2415



511
A L L F Q C V V D
15
2416



523
V K T I S I N Q V
15
2417



527
S I N Q V L S N K
15
2418



553
E V L K R E L G L
15
2419



558
E L C L A E C D I
15
2420



560
C L A E C D I I D
15
2421



582
A F F P D L V N M
15
2422



586
D L V N M L V L G
15
2423



592
V L G K H L C I P
15
2424



620
L L E P L C L H C
15
2425



623
P L G L H C T F I
15
2426



17
A V C V A G V H T
14
2427



70
F D A T L H I I V
14
2428



98
S S H F P L P L A
14
2429



101
E P L P L A Y A V
14
2430



115
V D I S L D C D L
14
2431



142
G P S C Y C G I L
14
2432



149
I L L V N C D R D
14
2433



169
Q H V H C L Q D L
14
2434



174
L Q D L E D M S V
14
2435



193
L F D D H K L V L
14
2436



208
A K R A Q V F H I
14
2437



227
R H V L G Q D K V
14
2438



231
G Q D K V S Y E V
14
2439



253
G L S F P D A G F
14
2440



256
F P D A G F T G L
14
2441



292
R V A P W I M T P
14
2442



297
I M T P S T L P P
14
2443



346
R W I Q D E M E L
14
2444



377
Q D F P Y K R I L
14
2445



383
R I L G P D F G Y
14
2446



438
S G R R V T Q V V
14
2447



455
V Q P P V E L F V
14
2448



466
L A V G H V D E F
14
2449



498
K L F Q S K Q K C
14
2450



517
V V D D E Q V K T
14
2451



530
Q V L S N K D L I
14
2452



587
L V N M L V L G K
14
2453



609
G C C C L E E K V
14
2454



618
R S L L S P L C L
14
2455



630
F I D D F T P Y H
14
2456



638
H M L H C E V H C
14
2457



55
N M E R G R E R A
13
2458



271
L L D D S N S D F
13
2459



282
S P I F T D T V V
13
2460



334
L T I C P Q A S N
13
2461



349
Q D E M S L G Y V
13
2462



362
K T L P V V F D S
13
2463



393
T R S P R D R S V
13
2464



403
C L D S F G N L E
13
2465



426
G R I L I C C N L
13
2466



427
R I L I G C N L P
13
2467



445
V V R D F L H A Q
13
2468



447
R D F L H A Q K V
13
2469



449
F L H A Q K V Q P
13
2470



488
M L L A S P C A C
13
2471



504
Q K C C H C R A L
13
2472



531
V L S N K D L I N
13
2473



571
Q L F K T S R K K
13
2474



574
K T E R K K A T A
13
2475



590
M L V L C K H L G
13
2476



599
I P K P F C P I I
13
2477



606
I I N C C C C L E
13
2478



621
L E P L G L H C T
13
2479



655
F S F K W W N M V
13
2480



32
S V P 5 G T S M F
12
2481



42
V Y C T P G V D I
12
2482



51
Y I S P N M E R G
12
2483



69
R F D A T L E I I
12
2484



84
S N D L N D S H V
12
2485



95
S Y H S S H E P L
12
2486



132
N F V D K R Q W V
12
2487



150
L L V N C D R D D
12
2488



173
C L Q D L E D M S
12
2489



182
V M V L R T Q G P
12
2490



217
C G P E D V C E A
12
2491



222
V C E A Y R H V L
12
2492



241
R L H G D E E R F
12
2493



243
H G D E E R F F V
12
2494



246
E E R F F V E G L
12
2495



257
P D A G F T C L I
12
2496



280
S A S P I F T D T
12
2497



294
A P W I M T P S T
12
2498



351
E M E L G Y V Q A
12
2499



363
T L P V V F D S P
12
2500



375
E L Q D F P Y K R
12
2501



396
P R D R S V S C L
12
2502



420
C K E Y P L C K I
12
2503



424
P L G R I L I G G
12
2504



433
N L P G S S C R R
12
2505



434
L P G S S G S R V
12
2506



454
K V Q P P V E L F
12
2507



474
F L S F V P A P D
12
2508



520
D E Q V K T I S I
12
2509



551
N R E V L K R S L
12
2510



559
L G L A E C D I I
12
2511



566
I I D I P Q L F K
12
2512



596
H L G I P K P F G
12
2513



632
D D F T P Y H M L
12
2514



5
R I V R V S L E H
11
2515



9
V S L E H P T S A
11
2516



97
H S S H E P L P L
11
2517



100
H E P L P L A Y A
11
2518



141
W G P 5 G Y C G I
11
2519



148
C I L L V N C D R
11
2520



155
D R D D P S C D V
11
2521



163
V Q D N C D Q H V
11
2522



215
H I C G P 5 D V C
11
2523



259
A G F T G L I S F
11
2524



295
P W I M T P S T L
11
2525



309
Y V C R V R N N T
11
2526



333
K L T I C P Q A E
11
2527



340
A E N R N D R W I
11
2528



411
E V S P P V V A N
11
2529



423
Y P L G R I L I G
11
2530



481
D G K G F R M L L
11
2531



526
I S I N Q V L S N
11
2532



58
F P D L V N M L V
11
2533



613
L E E K V R S L L
11
2534



625
G L H C T F I D D
11
2535



642
G E V H C G T N V
11
2536



6
I V R V S L E H P
10
2537



37
T E M F E V Y G T
10
2538



38
E M F E V Y G T P
10
2539



47
G V D I Y I S P N
10
2540



75
E I I V V M N S P
10
2541



76
I I V V M N S P S
10
2542



108
A V L Y L T C V D
10
2543



112
L T C V D I S L D
10
2544



122
D L N C E G R Q D
10
2545



172
H C L Q D L E D M
10
2546



183
M V L R T Q G P A
10
2547



237
Y E V P R L H G D
10
2548



279
F S A S P I F T D
10
2549



283
P I F T D T V V F
10
2550



296
W I M T P S T L P
10
2551



300
P S T L P P L E V
10
2552



325
E L A R K A G G E
10
2553



329
K A G C K L T I C
10
2554



335
T I C P Q A E N R
10
2555



352
M E L G Y V Q A P
10
2556



485
C F R M L L A S P
10
2557



490
L A S P G A C F E
10
2558



515
Q G V V D D E Q V
10
2559



525
T I S I N Q V L S
10
2560



537
L I N Y N K F V Q
10
2561



556
K R E L G L A E C
10
2562



565
D I I D I P Q L F
10
2563



585
P D L V N M L V L
10
2564



594
G K H L G I P K P
10
2565



636
P Y H N L H C E V
10
2566



12
E H P T S A V C V
9
2567



22
C V E T L V D I Y
9
2568



24
E T L V D I Y C S
9
2569



31
G S V P E G T E H
9
2570



34
P E G T E M F E V
9
2571



43
Y G T P G V D I Y
9
2572



49
D I Y I S P N M E
9
2573



77
I V V M N S P S N
9
2574



146
Y G G I L L V N C
9
2575



147
G G I L L V N C D
9
2576



186
R T Q G P A A L F
9
2577



195
D D H K L V L H T
9
2578



202
H T S S Y D A K R
9
2579



249
F F V E G L S F P
9
2580



254
L S F P D A G F T
9
2581



288
T V V F R V A P W
9
2582



316
N T C F V D A V A
9
2583



319
F V D A V A E L A
9
2584



321
D A V A E L A R K
9
2585



347
W I Q D E N E L G
9
2586



376
L Q D F P Y K R I
9
2587



385
L G P D F G Y V T
9
2588



400
S V S G L D S F G
9
2589



410
L E V S P P V V A
9
2590



412
V S P P V V A N G
9
2591



416
V V A N G K E Y P
9
2592



444
Q V V R D F L H A
9
2593



450
L H A Q K V Q P P
9
2594



461
L F V D W L A V G
9
2595



462
F V D W L A V G H
9
2596



480
A P D G K G F R M
9
2597



481
P D G K G F R M L
9
2598



494
G A C F K L F Q E
9
2599



529
N Q V L S N K D L
9
2600



543
F V Q S C I D W N
9
2601



568
D I P Q L F K T E
9
2602



601
K P F G P I I N G
9
2603



640
L H G E V H C G T
9
2604



20
V A G V E T L V D
8
2605



74
L E I I V V M N S
8
2606



106
A Y A V L Y L T C
8
2607



110
L Y L T C V D I S
8
2608



114
C V D I S L D C D
8
2609



126
E G R Q D R N F V
8
2610



139
W V W C P S G Y G
8
2611



143
P S C Y C C I L L
8
2612



145
G Y C G I L L V N
8
2613



175
Q D L E D M S V M
8
2614



181
S V M V L R T Q G
8
2615



194
F D D H K L V L H
8
2616



199
L V L H T S S Y D
8
2617



22
E A Y R H V L G Q
8
2618



230
L G Q D K V S Y E
8
2619



284
I F T D T V V F R
8
2620



353
E L G Y V Q A P H
8
2621



355
G Y V Q A P H K T
8
2622



379
F P Y K R I L G P
8
2623



392
V T R E P R D R S
8
2624



399
R S V S C L D S F
8
2625



425
L G R I L I G G N
8
2626



432
G N L P G S S G R
8
2627



477
F V P A P D G K G
8
2628



484
K G F R M L L A S
8
2629



513
L F Q G V V D D E
8
2630



533
S N K D L I N Y N
8
2631



539
N Y N K F V Q S C
8
2632



540
Y N K F V Q S C I
8
2633



588
V N M L V L G K H
8
2634



597
L G I P K P F G P
8
2635



7
V R V S L E H P T
7
2636



8
R V S L E H P T S
7
2637



13
H P T S A V C V A
7
2638



26
L V D I Y G S V P
7
2639



29
I Y G S V P E G T
7
2640



36
G T E M F E V Y G
7
2641



48
V D I Y I S P N M
7
2642



50
I Y I S P N M E R
7
2643



64
D T R R W R F D A
7
2644



82
S P S N D L N D S
7
2645



91
H V Q I S Y H S S
7
2646



93
Q I S Y H S S H E
7
2647



103
L P L A Y A V L Y
7
2648



116
D I S L D C D L N
7
2649



129
Q D R N F V D K R
7
2650



151
L V N C D R D D P
7
2651



228
H V L C Q D K V S
7
2652



250
F V E G L S F P D
7
2653



260
G F T G L I S F H
7
2654



262
T G L I S F H V T
7
2655



266
S F H V T L L D D
7
2656



269
V T L L D D S N E
7
2657



275
S N E D F S A S P
7
2658



276
N E D F S A S P I
7
2659



287
D T V V F R V A P
7
2660



303
L P P L S V Y V C
7
2661



314
R N N T C F V D A
7
2662



324
A E L A R K A G C
7
2663



332
C K L T I C P Q A
7
2664



366
V V F D S P R N G
7
2665



406
S F G N L E V S P
7
2666



422
E Y P L G R I L I
7
2667



436
G S S G R R V T Q
7
2668



441
R V T Q V V R D F
7
2669



532
L S N K D L I N Y
7
2670



549
D W N R E V L K R
7
2671



555
L K R E L G L A E
7
2672



562
A E C D I I D I P
7
2673



626
L H C T F I D D F
7
2674



628
C T F I D D F T P
7
2675



635
T P Y H M L H G E
7
2676



649
N V C R K P F S F
7
2677



4
Q R I V R V S L E
6
2678



11
L E H P T S A V C
6
2679



16
S A V C V A G V E
6
2680



27
V D I Y C S V P E
6
2681



78
V V M N S P S N D
6
2682



87
L N D S H V Q I S
6
2683



92
V Q I S Y H S S H
6
2684



94
I S Y H S S H E P
6
2685



99
S H E P L P L A Y
6
2686



117
I S L D C D L N C
6
2687



119
L D C D L N C E G
6
2688



123
L N C S G R Q D R
6
2689



180
M S V M V L R T Q
6
2690



211
A Q V F H I C G P
6
2691



216
I C G P E D V C E
6
2692



225
A Y R H V L G Q D
6
2693



251
V E G L S F P D A
6
2694



265
I S F H V T L L D
6
2695



267
F H V T L L D D S
6
2696



268
H V T L L D D S N
6
2697



291
F R V A P W I M T
6
2698



327
A R K A G C K L T
6
2699



344
N D R W I Q D E M
6
2700



348
I Q D E M E L G Y
6
2701



361
H K T L P V V F D
6
2702



386
G P D F G Y V T R
6
2703



405
D S F G N L E V S
6
2704



407
F G N L E V S P P
6
2705



440
R R V T Q V V R D
6
2706



459
V E L F V D W L A
6
2707



472
D E F L S F V P A
6
2708



473
E F L S F V P A P
6
2709



495
A C F K L F Q E K
6
2710



503
K Q K C G H G R A
6
2711



507
G H G R A L L F Q
6
2712



516
G V V D D E Q V K
6
2713



528
I N Q V L S N K D
6
2714



535
K D L I N Y N K F
6
2715



538
I N Y N K F V Q S
6
2716



572
L F K T E R K K A
6
2717



573
F K T E R K K A T
6
2718



581
T A F F P D L V N
6
2719



608
N G C C C L E E K
6
2720



611
C C L E E K V R S
6
2721



624
L G L H C T F I D
6
2722



631
I D D F T P Y H M
6
2723



643
E V H C G T N V C
6
2724



30
Y G S V P E C T E
5
2725



45
T P G V D I Y I S
5
2726



65
T R R W R F D A T
5
2727



133
F V D K R Q W V W
5
2728



140
V W C P S G Y G G
5
2729



170
H V H C L Q D L E
5
2730



196
D H K L V L H T S
5
2731



197
H K L V L H T S S
5
2732



204
S S Y D A K R A Q
5
2733



210
R A Q V F H I C G
5
2734



212
Q V F H I C G P E
5
2735



255
S F P D A C F T G
5
2736



273
D D S N E D F S A
5
2737



290
V F R V A P W I M
5
2738



293
V A P W I M T P S
5
2739



308
V Y V C R V R N N
5
2740



320
V D A V A E L A R
5
2741



330
A G C K L T I C P
5
2742



357
V Q A P H K T L P
5
2743



370
S P R N C E L Q D
5
2744



382
K R I L G P D F G
5
2745



387
P D F G Y V T R E
5
2746



391
Y V T R E P R D R
5
2747



430
I G G N L P G S S
5
2748



431
G G N L P C S S G
5
2749



469
G H V D E F L S F
5
2750



475
L S F V P A P D G
5
2751



476
S F V P A P D C K
5
2752



478
V P A P D G K G F
5
2753



483
G K G F R M L L A
5
2754



508
H G R A L L F Q G
5
2755



548
I D W N R E V L K
5
2756



550
W N R E V L K R E
5
2757



557
R E L G L A E C D
5
2758



569
I P Q L F K T E R
5
2759



602
P F G P I I N C C
5
2760



607
I N G C C C L E E
5
2761



63
F T P Y H M L H G
5
2762



64
V H C G T N V C R
5
2763



647
G T N V C R K P F
5
2764



1
P T S A V C V A G
4
2765



41
E V Y G T P G V D
4
2766



58
R G R E R A D T R
4
2767



62
R A D T R R W R F
4
2768



135
D K R Q W V W G P
4
2769



137
R Q W V W G P S G
4
2770



162
D V Q D N C D Q H
4
2771



171
V H C L Q D L E D
4
2772



188
Q G P A A L F D D
4
2773



189
G P A A L F D D H
4
2774



190
P A A L F D D H K
4
2775



206
Y D A K R A Q V F
4
2776



207
D A K R A Q V F H
4
2777



209
K R A Q V F H I C
4
2778



213
V F H I C C P E D
4
2779



235
V S Y E V P R L H
4
2780



248
R F F V E G L S F
4
2781



307
E V Y V C R V R N
4
2782



310
V C R V R N N T C
4
2783



317
T C F V D A V A E
4
2784



323
V A E L A R K A G
4
2785



331
G C K L T I C P Q
4
2786



343
R N D R W I Q D E
4
2787



350
D E M E L G Y V Q
4
2788



354
L G Y V Q A P H K
4
2789



367
V F D S P R N C E
4
2790



371
P R N G E L Q D F
4
2791



374
G E L Q D F P Y K
4
2792



381
Y K R I L C P D F
4
2793



413
S P P V V A N G K
4
2794



419
N G K E Y P L C R
4
2795



456
Q P P V E L F V D
4
2796



479
P A P D C K G F R
4
2797



499
L F Q E K Q K C G
4
2798



595
K H L G I P K P F
4
2799



615
E K V R S L L E P
4
2800



622
E P L G L H C T F
4
2801



645
H C G T N V C R K
4
2802



651
C R K P F S F K W
4
2803



52
I S P N M E R G R
3
2804



53
S P N M E R G R E
3
2805



56
M E R G R E R A D
3
2806



57
E R G R E R A D T
3
2807



85
N D L N D S H V Q
3
2808



89
D S H V Q I S Y H
3
2809



90
S H V Q I S Y H S
3
2810



96
Y H S S H E P L P
3
2811



138
Q W V W G P S G Y
3
2812



153
N C D R D D P S C
3
2813



158
D P S C D V Q D N
3
2814



160
S C D V Q D N C D
3
2815



187
T Q C P A A L F D
3
2816



223
C E A Y R H V L G
3
2817



226
Y R H V L G Q D K
3
2818



236
S Y E V P R L H G
3
2819



239
V P R L H G D E E
3
2820



240
P R L H G D E E R
3
2821



242
L H G D F E R F F
3
2822



245
D E E R F F V E G
3
2823



258
D A G F T G L I S
3
2824



274
D S N E D F S A S
3
2825



278
D F S A S R I F T
3
2826



286
T D T V V F R V A
3
2827



304
P P L E V Y V C R
3
2828



336
I C P Q A E N R N
3
2829



339
Q A E N R N D R W
3
2830



364
L P V V F D S P R
3
2831



372
R N G E L Q D F P
3
2832



418
A N G K E Y P L G
3
2833



435
P G S S G R R V T
3
2834



439
G R R V T Q V V R
3
2835



446
V R D F L H A Q K
3
2836



486
F R M L L A S P G
3
2837



506
C G H G R A L L F
3
2838



514
F Q C V V D D S Q
3
2839



522
Q V K T I S I N Q
3
2840



578
K K A T A F F P D
3
2841



604
G P I I N G C C C
3
2842



617
V R S L L E P L G
3
2843



629
T F I D D F T P Y
3
2844



637
Y H N L H C F V H
3
2845



653
K P F S F K W W N
3
2846



33
V P E G T E M F E
2
2847



54
P N M E R G R E R
2
2848



60
R E R A D T R R W
2
2849



80
M N S P S N D L N
2
2850



83
P S N D L N D S H
2
2851



88
N D S H V Q I S Y
2
2852



113
T C V D I S L D C
2
2853



121
C D L N C E G R Q
2
2854



127
G R Q D R N F V D
2
2855



128
R Q D R N F V D K
2
2856



134
V D K R Q W V W G
2
2857



152
V N C D R D D P S
2
2858



156
R D D P S C D V Q
2
2859



164
Q D N C D Q H V H
2
2860



165
D N C D Q H V H C
2
2861



201
L H T S S Y D A K
2
2862



203
T S S Y D A K R A
2
2863



218
G P E D V C E A Y
2
2864



233
D K V S Y E V P R
2
2865



238
E V P R L H G D E
2
2866



244
G D E E R F F V E
2
2867



272
L D D S N E D F S
2
2868



299
T P S T L P P L E
2
2869



313
V R N N T C F V D
2
2870



338
P Q A E N R N D R
2
2871



342
N R N D R W I Q D
2
2872



388
D F G Y V T R E P
2
2873



389
F G Y V T R E P R
2
2874



390
G Y V T R E P R D
2
2875



395
E P R D R S V S G
2
2876



401
V S C L D S F C N
2
2877



415
P V V A N G K E Y
2
2878



452
A Q K V Q P P V E
2
2879



457
P P V E L F V D W
2
2880



464
D W L A V C H V D
2
2881



496
C F K L F Q E K Q
2
2882



575
T E R K K A T A F
2
2883



593
L G K H L C I P K
2
2884



627
H C T F I D D F T
2
2885



650
V C R K P F S F K
2
2886



63
A D T R R W R F D
1
2887



81
N S P S N D L N D
1
2888



124
N C E C R Q D K N
1
2889



131
R N F V D K R Q W
1
2890



136
K R Q W V W G P S
1
2891



154
C D R D D P S C D
1
2892



157
D D P S C D V Q D
1
2893



252
E G L S F P D A G
1
2894



306
L E V Y V C R V R
1
2895



365
P V V F D S P R N
1
2896



468
V G H V D E F L S
1
2897



492
S P G A C F K L F
1
2898



493
P G A C F K L F Q
1
2899



497
F K L F Q E K Q K
1
2900



500
F Q E K Q K C G H
1
2901



521
E Q V K T I S I N
1
2902



534
N K D L I N Y N K
1
2903



541
N K F V Q S C I D
1
2904



542
K F V Q S C I D W
1
2905



544
V Q S C I D W N R
1
2906



545
Q S C I D W N R E
1
2907



552
R E V L K R E L G
1
2908



570
P Q L F K T E R K
1
2909



603
F G P I I N G C C
1
2910



633
D F T P Y H M L H
1
2911



641
H G E V H C G T N
1
2912



646
C G T N V C R K P
1
2913



648
T N V C R K P F S
1
2914



654
P F S F K W W N M
1
2915



656
S F K W W N M V P
1
2916



67
R W R F D A T L E
−1
2917



219
P E D V C E A Y R
−1
2918



232
Q D K V S Y E V P
−1
2919



277
E D F S A S P I F
−1
2920



360
P H K T L P V V F
−1
2921



369
D S P R N C F L Q
−1
2922



373
N G E L Q D F P Y
−1
2923



378
D F P Y K R I L G
−1
2924



414
P P V V A N G K E
−1
2925



502
E K Q K C G H G R
−1
2926



577
R K K A T A F F P
−1
2927



610
C C C L E E K V R
−1
2928



35
E G T E M F E V Y
−2
2929



130
D R N F V D K R Q
−2
2930



159
P S C D V Q D N C
−2
2931



220
E D V C E A Y R H
−2
2932



519
D D E Q V K T I S
−2
2933



600
P K P F G P I I N
−2
2934



652
R K P F S F K W W
−2
2935



247
E R F F V E G L S
−3
2936



341
E N R N D R W I Q
−3
2937



563
E C D I I D I P Q
−3
2938



614
E E K V R S L L E
−3
2939










184P1E2 v.2: HLA Peptide Scoring


Results A0201 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






7
T LA P L E V Y V
27
2940



3
M TP S T L A P L
20
2941



2
I MT P S T L A P
15
2942



1
W IM T P S T L A
14
2943



6
S TL A P L E V Y
13
2944



8
L AP L E V Y V C
11
2945



5
P ST L A P L E V
9
2946



9
A PL E V Y V C R
8
2947



4
T PS T L A P L E
2
2948










184P1E2 v.3: HLA Peptide Scoring


Results A0201 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






1
D EF L S F V P V
12
2949



3
F LS F V P V P D
12
2950



4
L SF V P V P D G
9
2951



6
F VP V P D G K G
8
2952



9
V PD G K G F R M
8
2953



2
E FL S F V P V P
5
2954



5
S FV P V P D G K
5
2955



8
P VP D G K G F R
4
2956



7
V PV P D G K G F
3
2957
















TABLE XXIV







184P1E2: HLA Peptide Scoring


Results A0202 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID














NO DATA
















TABLE XXV







184P1E2: HLA Peptide Scoring


Results A0203 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID














NO DATA




















TABLE XXVI









SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A3 9-mers SYFPEITHI












325
E L A R K A G C K
29
2958



511
A L L F Q G V V D
26
2959



198
K L V L H T S S Y
25
2960



516
G V V D D E Q V K
25
2961



566
I I D I P Q L F K
24
2962



5
R I V R V S L E H
23
2963



292
R V A P W I M T P
23
2964



527
S I N Q V L S N K
23
2965



587
L V N M L V L G K
23
2966



41
E V Y G T P G V D
22
2967



229
V L G Q D K V S Y
22
2968



383
R I L G P D F G Y
22
2969



428
I L I G G N L P G
22
2970



571
Q L F K T E R K K
22
2971



26
L V D I Y G S V P
21
2972



58
R G R E R A D T R
21
2973



128
R Q D R N F V D K
21
2974



263
G L I S F H V T L
21
2975



415
P V V A N G K E Y
21
2976



462
F V D W L A V G H
21
2977



489
L L A S P G A C F
21
2978



649
N V C R K P F S F
21
2979



17
A V C V A G V E T
20
2980



241
R L H C D E E R F
20
2981



283
P I F T D T V V F
20
2982



427
R I L I G G N L P
20
2983



454
K V Q P P V E L F
20
2984



8
R V S L E H P T S
19
2985



32
S V P E G T F M F
19
2986



108
A V L Y L T C V D
19
2987



109
V L Y L T C V D I
19
2988



307
E V Y V C R V R N
19
2989



441
R V T Q V V R D F
19
2990



449
F L H A Q K V Q P
19
2991



548
I D W N R E V L K
19
2992



619
S L L E P L G L H
19
2993



620
L L E P L G L H C
19
2994



2
S L Q R I V R V S
18
2995



86
D L N D S H V Q I
18
2996



102
P L P L A Y A V L
18
2997



162
D V Q D N C D Q H
18
2998



228
H V L G Q D K V S
18
2999



289
V V F R V A P W I
18
3000



312
R V R N N T C F V
18
3001



322
A V A E L A R K A
18
3002



335
T I C P Q A E N R
18
3003



353
E L G Y V Q A P H
18
3004



391
Y V T R E P R D R
18
3005



411
E V S P P V V A N
18
3006



444
Q V V R D F L H A
18
3007



490
L A S P G A C F K
18
3008



591
L V L G K H L G I
18
3009



103
L P L A Y A V L Y
17
3010



184
V L R T Q G P A A
17
3011



253
G L S F P D A G F
17
3012



302
T L P P L E V Y V
17
3013



354
L G Y V Q A P H K
17
3014



433
N L P G S S G R R
17
3015



465
W L A V G H V D E
17
3016



488
M L L A S P G A C
17
3017



553
E V L K R E L G L
17
3018



565
D I I D I P Q L F
17
3019



22
G V E T L V D I Y
16
3020



73
T L E I I V V M N
16
3021



122
D L N C E G R Q D
16
3022



133
F V D K R Q W V W
16
3023



148
G I L L V N C D R
16
3024



176
D L E D M S V M V
16
3025



181
S V M V L R T Q G
16
3026



183
M V L R T Q G P A
16
3027



186
R T Q G P A A L F
16
3028



248
R F F V E G L S F
16
3029



301
S T L P P L E V Y
16
3030



319
F V D A V A E L A
16
3031



333
K L T I C P Q A E
16
3032



356
Y V Q A P H K T L
16
3033



375
E L Q D F P Y K R
16
3034



384
I L G P D F G Y V
16
3035



400
S V S G L D S F G
16
3036



446
V R D F L H A Q K
16
3037



512
L L F Q G V V D D
16
3038



616
K V R S L L E P L
16
3039



643
E V H C G T N V C
16
3040



650
V C R K P F S F K
16
3041



6
I V R V S L E H P
15
3042



10
S L E H P T S A V
15
3043



19
C V A G V E T L V
15
3044



49
D I Y I S P N M E
15
3045



78
V V M N S P S N D
15
3046



99
S H E P L P L A Y
15
3047



199
L V L H T S S Y D
15
3048



271
L L D D S N E D F
15
3049



288
T V V F R V A P W
15
3050



309
Y V C R V R N N T
15
3051



321
D A V A E L A R K
15
3052



394
R E P R D R S V S
15
3053



409
N L E V S P P V V
15
3054



413
S P P V V A N G K
15
3055



439
G R R V T Q V V R
15
3056



445
V V R D F L H A Q
15
3057



474
F L S F V P A P D
15
3058



476
S F V P A P D G K
15
3059



497
F K L F Q E K Q K
15
3060



517
V V D D E Q V K T
15
3061



531
V L S N K D L I N
15
3062



538
I N Y N K F V Q S
15
3063



554
V L K R E L G L A
15
3064



586
D L V N M L V L G
15
3065



25
T L V D I Y G S V
14
3066



28
D I Y G S V P E G
14
3067



47
G V D I Y I S P N
14
3068



66
R R W R F D A T L
14
3069



72
A T L E I I V V M
14
3070



75
E I I V V M N S P
14
3071



77
I V V M N S P S N
14
3072



104
P L A Y A V L Y L
14
3073



138
Q W V W G P S G Y
14
3074



149
I L L V N C D R D
14
3075



192
A L F D D H K L V
14
3076



206
Y D A K R A Q V F
14
3077



215
H I C G P E D V C
14
3078



238
E V P R L H G D E
14
3079



328
R K A G C K L T I
14
3080



360
P H K T L P V V F
14
3081



386
G P D F G Y V T R
14
3082



416
V V A N G K E Y P
14
3083



432
G N L P G S S G R
14
3084



460
E L F V D W L A V
14
3085



467
A V G H V D E F L
14
3086



470
H V D E F L S F V
14
3087



506
C G H G R A L L F
14
3088



530
Q V L S N K D L I
14
3089



536
D L I N Y N K F V
14
3090



574
K T E R K K A T A
14
3091



593
L G K H L G I P K
14
3092



605
P I I N G C C C L
14
3093



612
C L E E K V R S L
14
3094



76
I I V V M N S P S
13
3095



91
H V Q I S Y H S S
13
3096



93
Q I S Y H S S H E
13
3097



111
Y L T C V D I S L
13
3098



139
W V W G P S G Y G
13
3099



212
Q V F H I C G P E
13
3100



226
Y R H V L G Q D K
13
3101



234
K V S Y E V P R L
13
3102



268
H V T L L D D S N
13
3103



270
T L L D D S N E D
13
3104



363
T L P V V F D S P
13
3105



370
S P R N G E L Q D
13
3106



403
G L D S F G N L E
13
3107



421
K E Y P L G R I L
13
3108



424
P L G R I L I G G
13
3109



436
G S S G R R V T Q
13
3110



477
F V P A P D G K G
13
3111



495
A C F K L F Q E K
13
3112



498
K L F Q E K Q K C
13
3113



522
Q V K T I S I N Q
13
3114



525
T I S I N Q V L S
13
3115



547
C I D W N R E V L
13
3116



570
P Q L F K T E R K
13
3117



575
T E R K K A T A F
13
3118



590
M L V L G K H L G
13
3119



630
F I D D F T P Y H
13
3120



3
L Q R I V R V S L
12
3121



92
V Q I S Y H S S H
12
3122



144
S G Y G G I L L V
12
3123



173
C L Q D L E D M S
12
3124



175
Q D L E D M S V M
12
3125



190
P A A L F D D H K
12
3126



207
D A K R A Q V F H
12
3127



250
F V E G L S F P D
12
3128



304
P P L E V Y V C R
12
3129



324
A E L A R K A G C
12
3130



348
I Q D E M E L G Y
12
3131



366
V V F D S P R N G
12
3132



374
G E L Q D F P Y K
12
3133



395
E P R D R S V S G
12
3134



399
R S V S G L D S F
12
3135



438
S G R R V T Q V V
12
3136



458
P V E L F V D W L
12
3137



464
D W L A V G H V D
12
3138



510
R A L L F Q G V V
12
3139



537
L I N Y N K F V Q
12
3140



560
G L A E C D I I D
12
3141



568
D I P Q L F K T E
12
3142



595
K H L G I P K P F
12
3143



606
I I N G C C C L E
12
3144



622
E P L G L H C T F
12
3145



623
P L G L H C T F I
12
3146



637
Y H M L H G E V H
12
3147



639
M L H G E V H C G
12
3148



35
E G T E M F E V Y
11
3149



43
Y G T P G V D I Y
11
3150



51
Y I S P N M E R G
11
3151



62
R A D T R R W R F
11
3152



67
R W R F D A T L E
11
3153



106
A Y A V L Y L T C
11
3154



114
C V D I S L D C D
11
3155



118
S L D C D L N C E
11
3156



150
L L V N C D R D D
11
3157



151
L V N C D R D D P
11
3158



156
R D D P S C D V Q
11
3159



201
L H T S S Y D A K
11
3160



205
S Y D A K R A Q V
11
3161



221
D V C E A Y R H V
11
3162



225
A Y R H V L G Q D
11
3163



235
V S Y E V P R L H
11
3164



259
A G F T G L I S F
11
3165



284
I F T D T V V F R
11
3166



295
P W I M T P S T L
11
3167



381
Y K R I L G P D F
11
3168



478
V P A P D G K G F
11
3169



526
I S I N Q V L S N
11
3170



534
N K D L I N Y N K
11
3171



543
F V Q S C I D W N
11
3172



555
L K R E L G L A E
11
3173



557
R E L G L A E C D
11
3174



558
E L G L A E C D I
11
3175



592
V L G K H L G I P
11
3176



596
H L G I P K P F G
11
3177



618
R S L L E P L G L
11
3178



629
T F I D D F T P Y
11
3179



4
Q R I V R V S L E
10
3180



11
L E H P T S A V C
10
3181



16
S A V C V A G V E
10
3182



50
I Y I S P N M E R
10
3183



54
P N M E R G R E R
10
3184



129
Q D R N F V D K R
10
3185



164
Q D N C D Q H V H
10
3186



170
H V H C L Q D L E
10
3187



200
V L H T S S Y D A
10
3188



224
E A Y R H V L G Q
10
3189



305
P L E V Y V C R V
10
3190



351
E M E L G Y V Q A
10
3191



365
P V V F D S P R N
10
3192



382
K R I L G P D F G
10
3193



419
N G K E Y P L G R
10
3194



429
L I G G N L P G S
10
3195



437
S S G R R V T Q V
10
3196



469
G H V D E F L S F
10
3197



484
K G F R M L L A S
10
3198



524
K T I S I N Q V L
10
3199



535
K D L I N Y N K F
10
3200



549
D W N R E V L K R
10
3201



577
R K K A T A F F P
10
3202



582
A F F P D L V N M
10
3203



608
N G C C C L E E K
10
3204



625
G L H C T F I D D
10
3205



644
V H C G T N V C R
10
3206



645
H C G T N V C R K
10
3207



27
V D I Y G S V P E
9
3208



60
R E R A D T R R W
9
3209



88
N D S H V Q I S Y
9
3210



101
E P L P L A Y A V
9
3211



116
D I S L D C D L N
9
3212



134
V D K R Q W V W G
9
3213



137
R Q W V W G P S G
9
3214



145
G Y G G I L L V N
9
3215



178
E D M S V M V L R
9
3216



187
T Q G P A A L F D
9
3217



194
F D D H K L V L H
9
3218



216
I C G P E D V C E
9
3219



218
G P E D V C E A Y
9
3220



255
S F P D A G F T G
9
3221



260
G F T G L I S F H
9
3222



264
L I S F H V T L L
9
3223



282
S P I F T D T V V
9
3224



296
W I M T P S T L P
9
3225



306
L E V Y V C R V R
9
3226



310
V C R V R N N T C
9
3227



320
V D A V A E L A R
9
3228



347
W I Q D E M E L G
9
3229



362
K T L P V V F D S
9
3230



379
F P Y K R I L G P
9
3231



389
F G Y V T R E P R
9
3232



397
R D R S V S G L D
9
3233



410
L E V S P P V V A
9
3234



423
Y P L G R I L I G
9
3235



431
G G N L P G S S G
9
3236



452
A Q K V Q P P V E
9
3237



480
A P D G K G F R M
9
3238



505
K C G H G R A L L
9
3239



556
K R E L G L A E C
9
3240



585
P D L V N M L V L
9
3241



598
G I P K P F G P I
9
3242



599
I P K P F G P I I
9
3243



610
C C C L E E K V R
9
3244



1
M S L Q R I V R V
8
3245



14
P T S A V C V A G
8
3246



18
V C V A G V E T L
8
3247



21
A G V E T L V D I
8
3248



56
M E R G R E R A D
8
3249



59
G R E R A D T R R
8
3250



61
E R A D T R R W R
8
3251



63
A D T R R W R F D
8
3252



117
I S L D C D L N C
8
3253



123
L N C E G R Q D R
8
3254



154
C D R D D P S C D
8
3255



193
L F D D H K L V L
8
3256



208
A K R A Q V F H I
8
3257



233
D K V S Y E V P R
8
3258



240
P R L H G D E E R
8
3259



281
A S P I F T D T V
8
3260



297
I M T P S T L P P
8
3261



311
C R V R N N T C F
8
3262



314
R N N T C F V D A
8
3263



316
N T C F V D A V A
8
3264



317
T C F V D A V A E
8
3265



327
A R K A G C K L T
8
3266



346
R W I Q D E M E L
8
3267



350
D E M E L G Y V Q
8
3268



385
L G P D F G Y V T
8
3269



406
S F G N L E V S P
8
3270



426
G R I L I G G N L
8
3271



440
R R V T Q V V R D
8
3272



447
R D F L H A Q K V
8
3273



461
L F V D W L A V G
8
3274



479
P A P D G K G F R
8
3275



485
G F R M L L A S P
8
3276



504
Q K C G H G R A L
8
3277



508
H G R A L L F Q G
8
3278



567
I D I P Q L F K T
8
3279



569
I P Q L F K T E R
8
3280



576
E R K K A T A F F
8
3281



597
L G I P K P F G P
8
3282



633
D F T P T H M L H
8
3283



638
H M L H G E V H C
8
3284



9
V S L E H P T S A
7
3285



12
E H P T S A V C V
7
3286



20
V A G V E T L V D
7
3287



31
G S V P E G T E M
7
3288



38
E M F E V Y G T P
7
3289



42
V Y G T P G V D I
7
3290



52
I S P N M E R G R
7
3291



57
E R G R E R A D T
7
3292



64
D T R R W R F D A
7
3293



83
P S N D L N D S H
7
3294



85
N D L N D S H V Q
7
3295



89
D S H V Q I S Y H
7
3296



105
L A Y A V L Y L T
7
3297



125
C E G R Q D R N F
7
3298



127
G R Q D R N F V D
7
3299



131
R N F V D K R Q W
7
3300



157
D D P S C D V Q D
7
3301



185
L R T Q G P A A L
7
3302



189
G P A A L F D D H
7
3303



196
D H K L V L H T S
7
3304



202
H T S S T D A K R
7
3305



204
S S Y D A K R A Q
7
3306



219
P E D V C E A Y R
7
3307



220
E D V C E A Y R H
7
3308



222
V C E A Y R H V L
7
3309



223
C E A Y R H V L G
7
3310



227
R H V L G Q D K V
7
3311



232
Q D K V S Y E V P
7
3312



239
V P R L H G D E E
7
3313



244
G D E E R F F V E
7
3314



245
D E E R F F V E G
7
3315



254
L S F P D A G F T
7
3316



262
T G L I S F H V T
7
3317



275
S N E D F S A S P
7
3318



338
P Q A E N R N D R
7
3319



340
A E N R N D R W I
7
3320



343
R N D R W I Q D E
7
3321



359
A P H K T L P V V
7
3322



364
L P V V F D S P R
7
3323



373
N G E L Q D F P Y
7
3324



393
T R E P R D R S V
7
3325



412
V S P P V V A N G
7
3326



430
I G G N L P G S S
7
3327



435
P G S S G R R V T
7
3328



453
Q K V Q P P V E L
7
3329



455
V Q P P V E L F V
7
3330



486
F R M L L A S P G
7
3331



487
R M L L A S P G A
7
3332



492
S P G A C F K L F
7
3333



502
E K Q K C G H G R
7
3334



503
K Q K C G H G R A
7
3335



552
R E V L K R E L G
7
3336



564
C D I I D I P Q L
7
3337



581
T A F F P D L V N
7
3338



588
V N M L V L G K H
7
3339



604
G P I I N G C C C
7
3340



611
C C L E E K V R S
7
3341



656
S F K W W N M V P
7
3342



55
N M E R G R E R A
6
3343



65
T R R W R F D A T
6
3344



69
R F D A T L E I I
6
3345



74
L E I I V V M N S
6
3346



94
I S Y H S S H E P
6
3347



140
V W G P S G Y G G
6
3348



142
G P S G Y G G I L
6
3349



168
D Q H V H C L Q D
6
3350



214
F H I C G P E D V
6
3351



249
F F V E G L S F P
6
3352



258
D A G F T G L I S
6
3353



265
I S F H V T L L D
6
3354



269
V T L L D D S N E
6
3355



276
N E D F S A S P I
6
3356



287
D T V V F R V A P
6
3357



294
A P W I M T P S T
6
3358



326
L A R K A G C K L
6
3359



334
L T I C P Q A E N
6
3360



342
N R N D R W I Q D
6
3361



358
Q A P H K T L P V
6
3362



368
F D S P R N G E L
6
3363



369
D S P R N G E L Q
6
3364



371
P R N G E L Q D F
6
3365



396
P R D R S V S G L
6
3366



398
D R S V S G L D S
6
3367



405
D S F G N L E V S
6
3368



443
T Q V V R D F L H
6
3369



448
D F L H A Q K V Q
6
3370



456
Q P P V E L F V D
6
3371



466
L A V G H V D E F
6
3372



507
G H G R A L L F Q
6
3373



532
L S N K D L I N Y
6
3374



614
E E K V R S L L E
6
3375



615
E K V R S L L E P
6
3376



30
Y G S V P E G T E
5
3377



71
D A T L E I I V V
5
3378



84
S N D L N D S H V
5
3379



120
D C D L N C E G R
5
3380



136
K R Q W V W G P S
5
3381



143
P S G Y G G I L L
5
3382



146
T G G I L L V N C
5
3383



153
N C D R D D P S C
5
3384



165
D N C D Q H V H C
5
3385



177
L E D M S V M V L
5
3386



180
M S V M V L R T Q
5
3387



191
A A L F D D H K L
5
3388



197
H K L V L H T S S
5
3389



209
K R A Q V F H I C
5
3390



230
L G Q D K V S Y E
5
3391



236
S Y E V P R L H G
5
3392



237
Y E V P R L H G D
5
3393



242
L H G D E E R F F
5
3394



256
F P D A G F T G L
5
3395



274
D S N E D F S A S
5
3396



279
F S A S P I F T D
5
3397



280
S A S P I F T D T
5
3398



286
T D T V V F R V A
5
3399



300
P S T L P P L E V
5
3400



303
L P P L E V Y V C
5
3401



313
V R N N T C F V D
5
3402



329
K A G C K L T I C
5
3403



332
C K L T I C P Q A
5
3404



339
Q A E N R N D R W
5
3405



341
E N R N D R W I Q
5
3406



352
M E L G Y V Q A P
5
3407



357
V Q A P H K T L P
5
3408



361
H K T L P V V F D
5
3409



372
R N G E L Q D F P
5
3410



377
Q D F P T K R I L
5
3411



380
P Y K R I L G P D
5
3412



392
V T R E P R D R S
5
3413



402
S G L D S F G N L
5
3414



404
L D S F G N L E V
5
3415



425
L G R I L I G G N
5
3416



457
P P V E L F V D W
5
3417



472
D E F L S F V P A
5
3418



482
D G K G F R M L L
5
3419



483
G K G F R M L L A
5
3420



491
A S P G A C F K L
5
3421



501
Q E K Q K C G H G
5
3422



515
Q G V V D D E Q V
5
3423



518
V D D E Q V K T I
5
3424



540
Y N K F V Q S C I
5
3425



546
S C I D W N R E V
5
3426



550
W N R E V L K R E
5
3427



579
K A T A F F P D L
5
3428



580
A T A F F P D L V
5
3429



589
N M L V L G K H L
5
3430



601
K P F G P I I N G
5
3431



607
I N G C C C L E E
5
3432



613
L E E K V R S L L
5
3433



628
C T F I D D F T P
5
3434



641
H G E V H C G T N
5
3435



647
G T N V C R K P F
5
3436



652
R K P F S F K W W
5
3437



13
H P T S A V C V A
4
3438



15
T S A V C V A G V
4
3439



29
I Y G S V P E G T
4
3440



36
G T E M F E V Y G
4
3441



39
M F E V T G Y P G
4
3442



68
W R F D A T L E I
4
3443



97
H S S H E P L P L
4
3444



100
H E P L P L A Y A
4
3445



113
T C V D I S L D C
4
3446



121
C D L N C E G R Q
4
3447



124
N C E G R Q D R N
4
3448



126
E G R Q D R N F V
4
3449



132
N F V D K R Q W V
4
3450



147
G G I L L V N C D
4
3451



171
V H C L Q D L E D
4
3452



174
L Q D L E D M S V
4
3453



188
Q G P A A L F D D
4
3454



210
R A Q V F H I C G
4
3455



213
V F H I C G P E D
4
3456



252
E G L S F P D A G
4
3457



257
P D A G F T G L I
4
3458



266
S F H V T L L D D
4
3459



277
E D F S A S P I F
4
3460



290
V F R V A P W I M
4
3461



318
C F V D A V A E L
4
3462



407
F G N L E V S P P
4
3463



408
G N L E V S P P V
4
3464



422
E Y P L G R I L I
4
3465



471
V D E F L S F V P
4
3466



493
P G A C F K L F Q
4
3467



494
G A C F K L F Q E
4
3468



500
F Q E K Q K C G H
4
3469



523
V K T I S I N Q V
4
3470



544
V Q S C I D W N R
4
3471



559
L G L A E C D I I
4
3472



562
A E C D I I D I P
4
3473



573
F K T E R K K A T
4
3474



578
K K A T A F F P D
4
3475



626
L H C T F I D D F
4
3476



635
T P Y H M L H G N
4
3477



642
G E V H C G T N V
4
3478



651
C R K P F S F K W
4
3479



653
K P F S F K W W N
4
3480



33
V P E G T E M F E
3
3481



44
G T P G V D I Y I
3
3482



46
P G V D I Y I S P
3
3483



48
V D I Y I S P N M
3
3484



53
S P N M E R G R E
3
3485



80
M N S P S N D L N
3
3486



81
N S P S N D L N D
3
3487



112
L T C V D I S L D
3
3488



115
V D I S L D C D L
3
3489



135
D K R Q W V W G P
3
3490



141
W G P S G Y G G I
3
3491



152
V N C D R D D P S
3
3492



179
D M S V M V L R T
3
3493



182
V H V L R T Q G P
3
3494



195
D D H K L V L H T
3
3495



203
T S S T D A K R A
3
3496



211
A Q V F H I C G P
3
3497



246
E E R F F V E G L
3
3498



273
D D S N E D F S A
3
3499



278
D F S A S P I F T
3
3500



291
F R V A P W I M T
3
3501



323
V A E L A R K A G
3
3502



330
A G C K L T I C P
3
3503



344
N D R W I Q D E M
3
3504



349
Q D E M E L G Y V
3
3505



387
P D F G Y V T R E
3
3506



417
V A N G K E Y P L
3
3507



418
A N G K E Y P L G
3
3508



434
L P G S S G R R V
3
3509



459
V E L F V D W L A
3
3510



473
E F L S F V P A P
3
3511



475
L S F V P A P D G
3
3512



481
P D G K G F R M L
3
3513



513
L F Q G V V D D E
3
3514



520
D E Q V K T I S I
3
3515



521
E Q V K T I S I N
3
3516



533
S N K D L I N Y N
3
3517



542
K F V Q S C I D W
3
3518



545
Q S C I D W N R E
3
3519



584
F P D L V N M L V
3
3520



594
G K H L G I P K P
3
3521



600
P K P F G P I I N
3
3522



602
P F G P I I N G C
3
3523



621
L E P L G L H C T
3
3524



631
I D D F T P Y H M
3
3525



634
F T P Y H M L H G
3
3526



23
V E T L V D I Y G
2
3527



24
E T L V D I Y G S
2
3528



40
F E V Y G T P G V
2
3529



82
S P S N D L N D S
2
3530



98
S S H E P L P L A
2
3531



107
Y A V L Y L T C V
2
3532



110
L Y L T C V D I S
2
3533



159
P S C D V Q D N C
2
3534



169
Q H V H C L Q D L
2
3535



172
H C L Q D L E D M
2
3536



243
H G D E E R F F V
2
3537



247
E R F F V E G L S
2
3538



308
V Y V C R V R N N
2
3539



331
G C K L T I C P Q
2
3540



336
I C P Q A E N R N
2
3541



337
C P Q A E N R N D
2
3542



345
D R W I Q D E M E
2
3543



378
D F P T K R I L G
2
3544



390
G Y V T R S P R D
2
3545



442
V T Q V V R D F L
2
3546



450
L H A Q K V Q P P
2
3547



463
V D W L A V G H V
2
3548



496
C F K L F Q E K Q
2
3549



499
L F Q S K Q K C G
2
3550



509
G R A L L F Q G V
2
3551



519
D D E Q V K T I S
2
3552



528
I N Q V L S N K D
2
3553



539
N Y N K F V Q S C
2
3554



572
L F K T E R K K A
2
3555



583
F F P D L V N M L
2
3556



603
F G P I I N G C C
2
3557



624
L G L H C T F I D
2
3558



640
L H G E V H C G T
2
3559



646
C G T N V C R K P
2
3560



648
T N V C R K P F S
2
3561



70
F D A T L E I I V
1
3562



90
S H V Q I S Y H S
1
3563



95
S Y H S S H E P L
1
3564



119
L D C D L N C E G
1
3565



130
D R N F V D K R Q
1
3566



155
D R D D P S C D V
1
3567



160
S C D V Q D N C D
1
3568



163
V Q D N C D Q H V
1
3569



167
C D Q H V H C L Q
1
3570



217
C G P E D V C E A
1
3571



231
G Q D K V S Y E V
1
3572



251
V E G L S F P D A
1
3573



261
F T G L I S F H V
1
3574



285
F T D T V V F R V
1
3575



293
V A P W I M T P S
1
3576



299
T P S T L P P L E
1
3577



315
N N T C F V D A V
1
3578



355
G Y V Q A P H K T
1
3579



367
V F D S P R N G E
1
3580



388
D F G Y V T R E P
1
3581



401
V S G L D S F G N
1
3582



414
P P V V A N G K E
1
3583



420
G K E Y P L G R I
1
3584



451
H A Q K V Q P P V
1
3585



514
F Q G V V D D E Q
1
3586



529
N Q V L S N K D L
1
3587



563
E C D I I D I P Q
1
3588



617
V R S L L E P L G
1
3589



632
D D F T P Y H M L
1
3590



636
P Y H M L H G E V
1
3591



654
P F S F K W W N M
1
3592



655
F S F K W W N M V
1
3593







184P1E2 v.2: HLA Peptide


Scoring Results A3 9-mers SYFPEITHI












7
T L A P L E V Y V
18
3594



6
S T L A P L E V Y
15
3595



9
A P L E V Y V C R
15
3596



1
W I M T P S T L A
9
3597



2
I M T P S T L A P
9
3598



5
P S T L A P L E V
5
3599



8
L A P L E V Y V C
5
3600



3
M T P S T L A P L
3
3601



4
T P S T L A P L E
3
3602







184P1E2 v.3: HLA Peptide


Scoring Results A3 9-mers SYFPEITHI












8
P V P D G K G F R
18
3603



3
F L S F V P V P D
16
3604



5
S F V P V P D G K
16
3605



6
F V P V P D G K G
11
3606



7
V P V P D G K G F
10
3607



2
E F L S F V P V P
6
3608



9
V P D G K G F R M
6
3609



1
D E F L S F V P V
5
3610




















TABLE XXVII









SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A26 9-mers SYFPEITHI












565
D I I D I P Q L F
30
3611



301
S T L P P L E V Y
26
3612



454
K V Q P P V E L F
26
3613



553
E V L K R E L G L
26
3614



22
G V E T L V D I Y
25
3615



28
D I Y G S V P E G
25
3616



72
A T L E I I V V M
25
3617



298
M T P S T L P P L
25
3618



441
R V T Q V V R D F
25
3619



32
S V P E G T E M F
24
3620



234
K V S Y E V P R L
24
3621



411
E V S P P V V A N
24
3622



582
A F F P D L V N M
24
3623



612
C L E E K V R S L
24
3624



616
K V R S L L E P L
24
3625



75
E I I V V M N S P
23
3626



186
R T Q G P A A L F
23
3627



221
D V C E A Y R H V
23
3628



283
P I F T D T V V F
23
3629



383
R I L G P D F G Y
23
3630



568
D I P Q L F K T E
23
3631



629
T F I D D F T P Y
23
3632



35
E G T E M F E V Y
22
3633



102
P L P L A Y A V L
22
3634



264
L I S F H V T L L
22
3635



458
P V E L F V D W L
22
3636



524
K T I S I N Q V L
22
3637



583
F F P D L V N M L
22
3638



649
N V C R K P F S F
22
3639



24
E T L V D I Y G S
21
3640



104
P L A T A V L Y L
21
3641



229
V L G Q D K V S Y
21
3642



241
R L H G D E E R F
21
3643



271
L L D D S N E D F
21
3644



318
C F V D A V A E L
21
3645



415
P V V A N G K E Y
21
3646



489
L L A S P G A C F
21
3647



632
D D F T P Y H M L
21
3648



198
K L V L H T S S Y
20
3649



253
G L S F P D A G F
20
3650



263
G L I S F H V T L
20
3651



277
E D F S A S P I F
20
3652



605
P I I N G C C C L
20
3653



647
G T N V C R K P F
20
3654



246
E E R F F V E G L
19
3655



248
R F F V E G L S F
19
3656



473
E F L S F V P A P
19
3657



547
C I D W N R E V L
19
3658



576
E R K K A T A F F
19
3659



586
D L V N M L V L G
19
3660



622
E P L G L H C T F
19
3661



41
E V Y G T P G V D
18
3662



49
D I Y I S P N M E
18
3663



86
D L N D S H V Q I
18
3664



162
D V Q D N C D Q H
18
3665



238
E V P R L H G D E
18
3666



287
D T V V F R V A P
18
3667



292
R V A P W I M T P
18
3668



307
E V Y V C R V R N
18
3669



356
T V Q A P H K T L
18
3670



429
L I G G N L P G S
18
3671



442
V T Q V V R D F L
18
3672



467
A V G H V D E F L
18
3673



470
H V D E F L S F V
18
3674



482
D G K G F R M L L
18
3675



51
Y I S P N M E R G
17
3676



111
Y L T C V D I S L
17
3677



176
D L E D M S V M V
17
3678



259
A G F T G L I S F
17
3679



371
P R N G E L Q D F
17
3680



460
E L F V D W L A V
17
3681



466
L A V G H V D E F
17
3682



527
S I N Q V L S N K
17
3683



6
I V R V S L E H P
16
3684



43
Y G T P G V D I Y
16
3685



47
G V D I Y I S P N
16
3686



64
D T R R W R F D A
16
3687



91
H V Q I S T H S S
16
3688



114
C V D I S L D C D
16
3689



116
D I S L D C D L N
16
3690



193
L F D D H K L V L
16
3691



285
F T D T V V F R V
16
3692



325
E L A R K A G C K
16
3693



362
K T L P V V F D S
16
3694



375
E L Q D F P Y K R
16
3695



396
P R D R S V S G L
16
3696



399
R S V S G L D S F
16
3697



445
V V R D F L H A Q
16
3698



469
G H V D E F L S F
16
3699



512
L L F Q G V V D D
16
3700



532
L S N K D L I N Y
16
3701



536
D L I N Y N K F V
16
3702



564
C D I I D I P Q L
16
3703



587
L V N M L V L G K
16
3704



598
G I P K P F G P I
16
3705



634
F T P Y H M L H G
16
3706



643
E V H C G T N V C
16
3707



654
P F S F K W W N M
16
3708



112
L T C V D I S L D
15
3709



122
D L N C E G R Q D
15
3710



175
Q D L E D M S V M
15
3711



177
L E D M S V M V L
15
3712



288
T V V F R V A P W
15
3713



322
A V A E L A R K A
15
3714



347
W I Q D E M E L G
15
3715



353
E L G Y V Q A P H
15
3716



366
V V F D S P R N G
15
3717



402
S G L D S F G N L
15
3718



444
Q V V R D F L H A
15
3719



535
K D L I N Y N K F
15
3720



543
F V Q S C I D W N
15
3721



633
D F T P Y H M L H
15
3722



25
T L V D I Y G S V
14
3723



38
E M F E V Y G T P
14
3724



73
T L E I I V V M N
14
3725



99
S H E P L P L A Y
14
3726



166
N C D Q H V H C L
14
3727



256
F P D A G F T G L
14
3728



260
G F T G L I S F H
14
3729



274
D S N E D F S A S
14
3730



290
V F R V A P W I M
14
3731



334
L T I C P Q A E N
14
3732



335
T I C P Q A E N R
14
3733



348
I Q D E M E L G Y
14
3734



363
T L P V V F D S P
14
3735



378
D F P Y K R I L G
14
3736



384
I L G P D F G Y V
14
3737



400
S V S G L D S F G
14
3738



405
D S F G N L E V S
14
3739



462
F V D W L A V G H
14
3740



472
D E F L S F V P A
14
3741



478
V P A P D G K G F
14
3742



481
P D G K G F R M L
14
3743



492
S P G A C F K L F
14
3744



517
V V D D E Q V K T
14
3745



558
E L G L A E C D I
14
3746



591
L V L G K H L G I
14
3747



619
S L L E P L G L H
14
3748



626
L H C T F I D D F
14
3749



639
M L H G E V H C G
14
3750



19
C V A G V E T L V
13
3751



44
G T P G V D I Y I
13
3752



78
V V M N S P S N D
13
3753



118
S L D C D L N C E
13
3754



169
Q H V H C L Q D L
13
3755



172
H C L Q D L E D M
13
3756



195
D D H K L V L H T
13
3757



202
H T S S Y D A K R
13
3758



206
Y D A K R A Q V F
13
3759



228
H V L G Q D K V S
13
3760



249
F F V E G L S F P
13
3761



250
F V E G L S F P D
13
3762



278
D F S A S P I F T
13
3763



289
V V F R V A P W I
13
3764



302
T L P P L E V Y V
13
3765



305
P L E V Y V C R V
13
3766



346
R W I Q D F M E L
13
3767



360
P H K T L P V V F
13
3768



388
D F G Y V T R E P
13
3769



392
V T R E P R D R S
13
3770



424
P L G R I L I G G
13
3771



433
N L P G S S G R R
13
3772



480
A P D G K G F R M
13
3773



485
G F R M L L A S P
13
3774



498
K L F Q E K Q K C
13
3775



554
V L K R E L G L A
13
3776



575
T E R K K A T A F
13
3777



579
K A T A F F P D L
13
3778



592
V L G K H L G I P
13
3779



602
P F G P I I N G C
13
3780



630
F I D D F T P Y H
13
3781



2
S L Q R I V R V S
12
3782



14
P T S A V C V A G
12
3783



17
A V C V A G V E T
12
3784



18
V C V A G V E T L
12
3785



26
L V D I Y G S V P
12
3786



48
V D I Y I S P N M
12
3787



62
R A D T R R W R F
12
3788



77
I V V M N S P S N
12
3789



89
D S H V Q I S Y H
12
3790



93
Q I S Y H S S H E
12
3791



115
V D I S L D C D L
12
3792



125
C E G R Q D R N F
12
3793



133
F V D K R Q W V W
12
3794



135
D K R Q W V W G P
12
3795



138
Q W V W G P S G Y
12
3796



139
W V W G P S G Y G
12
3797



178
E D M S V M V L R
12
3798



181
S V M V L R T Q G
12
3799



212
Q V F H I C G P E
12
3800



215
H I C G P E D V C
12
3801



218
G P E D V C E A Y
12
3802



245
D E E R F F V E G
12
3803



268
H V T L L D D S N
12
3804



269
V T L L D D S N E
12
3805



284
I F T D T V V F R
12
3806



321
D A V A E L A R K
12
3807



351
E M E L G Y V Q A
12
3808



365
P V V F D S P R N
12
3809



377
Q D F P Y K R I L
12
3810



381
Y K R I L G P D F
12
3811



428
I L I G G N L P G
12
3812



448
D F L H A Q K V Q
12
3813



461
L F V D W L A V G
12
3814



477
F V P A P D G K G
12
3815



491
A S P G A C F K L
12
3816



506
C G H G R A L L F
12
3817



513
L F Q G V V D D E
12
3818



516
G V V D D E Q V K
12
3819



521
E Q V K T I S I N
12
3820



522
Q V K T I S I N Q
12
3821



571
Q L F K T E R K K
12
3822



574
K T E R K K A T A
12
3823



585
P D L V N M L V L
12
3824



595
K H L G I P K P F
12
3825



606
I I N G C C C L E
12
3826



615
E K V R S L L E P
12
3827



620
L L E P L G L H C
12
3828



625
G L H C T F I D D
12
3829



628
C T F I D D F T P
12
3830



5
R I V R V S L E H
11
3831



8
R V S L E H P T S
11
3832



31
G S V P E G T E M
11
3833



36
G T E M F E V Y G
11
3834



69
R F D A T L E I I
11
3835



76
I I V V M N S P S
11
3836



88
N D S H V Q I S Y
11
3837



103
L P L A Y A V L Y
11
3838



108
A V L Y L T C V D
11
3839



158
D P S C D V Q D N
11
3840



170
H V H C L Q D L E
11
3841



173
C L Q D L E D M S
11
3842



179
D M S V M V L R T
11
3843



184
V L R T Q G P A A
11
3844



185
L R T Q G P A A L
11
3845



192
A L F D D H K L V
11
3846



196
D H K L V L H T S
11
3847



199
L V L H T S S Y D
11
3848



224
E A Y R H V L G Q
11
3849



242
L H G D E E R F F
11
3850



261
F T G L I S F H V
11
3851



295
P W I M T P S T L
11
3852



309
Y V C R V R N N T
11
3853



316
N T C F V D A V A
11
3854



319
F V D A V A E L A
11
3855



368
F D S P R N G E L
11
3856



373
N G E L Q D F P Y
11
3857



416
V V A N G K E Y P
11
3858



417
V A N G K E Y P L
11
3859



421
K E Y P L G R I L
11
3860



426
G R I L I G G N L
11
3861



449
F L H A Q K V Q P
11
3862



453
Q K V Q P P V E L
11
3863



504
Q K C G H G R A L
11
3864



525
T I S I N Q V L S
11
3865



530
Q V L S N K D L I
11
3866



549
D W N R E V L K R
11
3867



560
G L A E C D I I D
11
3868



566
I I D I P Q L F K
11
3869



580
A T A F F P D L V
11
3870



3
L Q R I V R V S L
10
3871



10
S L E H P T S A V
10
3872



71
D A T L E I I V V
10
3873



95
S Y H S S H E P L
10
3874



101
E P L P L A Y A V
10
3875



142
G P S G Y G G I L
10
3876



148
G I L L V N C D R
10
3877



151
L V N C D R D D P
10
3878



157
D D P S C D V Q D
10
3879



165
D N C D Q H V H C
10
3880



183
M V L R T Q G P A
10
3881



266
S F H V T L L D D
10
3882



270
T L L D D S N E D
10
3883



296
W I M T P S T L P
10
3884



311
C R V R N N T C F
10
3885



312
R V R N N T C F V
10
3886



326
L A R K A G C K L
10
3887



333
K L T I C P Q A E
10
3888



391
Y V T R E P R D R
10
3889



403
G L D S F G N L E
10
3890



409
N L E V S P P V V
10
3891



427
R I L I G G N L P
10
3892



465
W L A V G H V D E
10
3893



488
M L L A S P G A C
10
3894



495
A C F K L F Q E K
10
3895



511
A L L F Q G V V D
10
3896



537
L I N Y N K F V Q
10
3897



551
N R E V L K R E L
10
3898



567
I D I P Q L F K T
10
3899



589
N M L V L G K H L
10
3900



613
L E E K V R S L L
10
3901



631
I D D F T P Y H M
10
3902



12
E H P T S A V C V
9
3903



66
R R W R F D A T L
9
3904



79
V M N S P S N D L
9
3905



109
V L Y L T C V D I
9
3906



132
N F V D K R Q W V
9
3907



149
I L L V N C D R D
9
3908



191
A A L F D D H K L
9
3909



200
V L H T S S Y D A
9
3910



344
N D R W I Q D E M
9
3911



350
D E M E L G Y V Q
9
3912



367
V F D S P R N G E
9
3913



387
P D F G Y V T R E
9
3914



395
E P R D R S V S G
9
3915



406
S F G N L E V S P
9
3916



412
V S P P V V A N G
9
3917



474
F L S F V P A P D
9
3918



476
S F V P A P D G K
9
3919



499
L F Q E K Q K C G
9
3920



505
K C G H G R A L L
9
3921



529
N Q V L S N K D L
9
3922



531
V L S N K D L I N
9
3923



596
H L G I P K P F G
9
3924



601
K P F G P I I N G
9
3925



618
R S L L E P L G L
9
3926



61
E R A D T R R W R
8
3927



74
L E I I V V M N S
8
3928



97
H S S H E P L P L
8
3929



120
D C D L N C E G R
8
3930



143
P S G Y G G I L L
8
3931



150
L L V N C D R D D
8
3932



155
D R D D P S C D V
8
3933



207
D A K R A Q V F H
8
3934



213
V F H I C G P E D
8
3935



217
C G P E D V C E A
8
3936



222
V C E A Y R H V L
8
3937



233
D K V S Y E V P R
8
3938



247
E R F F V E G L S
8
3939



252
E G L S F P D A G
8
3940



255
S F P D A G F T G
8
3941



345
D R W I Q D E M E
8
3942



352
M E L C Y V Q A P
8
3943



450
L H A Q K V Q P P
8
3944



457
P P V E L F V D W
8
3945



496
C F K L F Q E K Q
8
3946



502
E K Q K C G H G R
8
3947



509
G R A L L F Q G V
8
3948



518
V D D E Q V K T I
8
3949



520
D E Q V K T I S I
8
3950



526
I S I N Q V L S N
8
3951



542
K F V Q S C I D W
8
3952



572
L F K T E R K K A
8
3953



590
M L V L G K H L G
8
3954



623
P L G L H C T F I
8
3955



656
S F K W W N M V P
8
3956



1
M S L Q R I V R V
7
3957



4
Q R I V R V S L E
7
3958



15
T S A V C V A G V
7
3959



21
A G V E T L V D I
7
3960



57
E R G R E R A D T
7
3961



87
L N D S H V Q I S
7
3962



98
S S H E P L P L A
7
3963



126
E G R Q D R N F V
7
3964



130
D R N F V D K R Q
7
3965



141
W G P S G Y G G I
7
3966



146
T G G I L L V N C
7
3967



147
G G I L L V N C D
7
3968



168
D Q H V H C L Q D
7
3969



194
F D D H K L V L H
7
3970



201
L H T S S Y D A K
7
3971



220
E D V C E A Y R H
7
3972



230
L G Q D K V S Y E
7
3973



237
Y E V P R L H G D
7
3974



273
D D S N E D F S A
7
3975



304
P P L E V Y V C R
7
3976



369
D S P R N G E L Q
7
3977



398
D R S V S G L D S
7
3978



422
E Y P L G R I L I
7
3979



440
R R V T Q V V R D
7
3980



484
K G F R M L L A S
7
3981



519
D D E Q V K T I S
7
3982



533
S N K D L I N Y N
7
3983



538
I N Y N K F V Q S
7
3984



539
N Y N K F V Q S C
7
3985



550
W N R E V L K R E
7
3986



563
E C D I I D I P Q
7
3987



614
E E K V R S L L E
7
3988



39
M F E V Y G T P G
6
3989



46
P G V D I Y I S P
6
3990



100
H E P L P L A Y A
6
3991



106
A Y A V L Y L T C
6
3992



128
R Q D R N F V D K
6
3993



144
S G Y G G I L L V
6
3994



145
G Y G G I L L V N
6
3995



189
G P A A L F D D H
6
3996



208
A K R A Q V F H I
6
3997



209
K R A Q V F H I C
6
3998



232
Q D K V S Y E V P
6
3999



244
G D E E R F F V E
6
4000



251
V E G L S F P D A
6
4001



258
D A G F T G L I S
6
4002



279
F S A S P I F T D
6
4003



280
S A S P I F T D T
6
4004



341
E N R N D R W I Q
6
4005



343
R N D R W I Q D E
6
4006



359
A P H K T L P V V
6
4007



374
G E L Q D F P Y K
6
4008



376
L Q D F P Y K R I
6
4009



379
F P Y K R I L G P
6
4010



386
G P D F G Y V T R
6
4011



418
A N G K E Y P L G
6
4012



423
Y P L G R I L I G
6
4013



425
L G R I L I G G N
6
4014



46
D W L A V G H V D
6
4015



494
G A C F K L F Q E
6
4016



508
H G R A L L F Q G
6
4017



523
V K T I S I N Q V
6
4018



556
K R E L G L A E C
6
4019



594
G K H L G I P K P
6
4020



597
L G I P K P F G P
6
4021



650
V C R K P F S F K
6
4022



652
R K P F S F K W W
6
4023



13
H P T S A V C V A
5
4024



34
P E G T E M F E V
5
4025



37
T E M F E V Y G T
5
4026



50
I Y I S P N M E R
5
4027



65
T R R W R F D A T
5
4028



92
V Q I S Y H S S H
5
4029



129
Q D R N F V D K R
5
4030



188
Q G P A A L F D D
5
4031



211
A Q V F H I C G P
5
4032



225
A Y R H V L G Q D
5
4033



262
T G L I S F H V T
5
4034



267
F H V T L L D D S
5
4035



293
V A P W I M T P S
5
4036



303
L P P L E V Y V C
5
4037



308
V Y V C R V R N N
5
4038



314
R N N T C F V D A
5
4039



315
N N T C F V D A V
5
4040



329
K A G C K L T I C
5
4041



331
G C K L T I C P Q
5
4042



361
H K T L P V V F D
5
4043



380
P Y K R I L G P D
5
4044



407
F G N L E V S P P
5
4045



420
G K E Y P L G R I
5
4046



437
S S G R R V T Q V
5
4047



456
Q P P V E L F V D
5
4048



463
V D W L A V G H V
5
4049



562
A E C D I I D I P
5
4050



578
K K A T A F F P D
5
4051



588
V N M L V L G K H
5
4052



608
N G C C C L E E K
5
4053



621
L E P L G L H C T
5
4054



635
T P Y H M L H G E
5
4055



645
H C G T N V C R K
5
4056



45
T P G V D I Y I S
4
4057



68
W R F D A T L E I
4
4058



82
S P S N D L N D S
4
4059



105
L A Y A V L Y L T
4
4060



107
Y A V L Y L T C V
4
4061



110
L Y L T C V D I S
4
4062



140
V W G P S G Y G G
4
4063



156
R D D P S C D V Q
4
4064



180
M S V M V L R T Q
4
4065



205
S Y D A K R A Q V
4
4066



254
L S F P D A G F T
4
4067



286
T D T V V F R V A
4
4068



317
T C F V D A V A E
4
4069



349
Q D E M E L G Y V
4
4070



419
N G K E Y P L G R
4
4071



432
G N L P G S S G R
4
4072



446
V R D F L H A Q K
4
4073



447
R D F L H A Q K V
4
4074



483
G K G F R M L L A
4
4075



501
Q E K Q K C G H G
4
4076



507
G H G R A L L F Q
4
4077



546
S C I D W N R E V
4
4078



555
L K R E L G L A E
4
4079



561
L A E C D I I D I
4
4080



599
I P K P F G P I I
4
4081



611
C C L E E K V R S
4
4082



651
C R K P F S F K W
4
4083



655
F S F K W W N M V
4
4084



9
V S L E H P T S A
3
4085



11
L E H P T S A V C
3
4086



27
V D I Y G S V P E
3
4087



33
V P E G T E M F E
3
4088



54
P N M E R G R E R
3
4089



56
M E R G R E R A D
3
4090



80
M N S P S N D L N
3
4091



81
N S P S N D L N D
3
4092



83
P S N D L N D S H
3
4093



84
S N D L N D S H V
3
4094



119
L D C D L N C E G
3
4095



123
L N C E G R Q D R
3
4096



124
N C E G R Q D R N
3
4097



131
R N F V D K R Q W
3
4098



134
V D K R Q W V W G
3
4099



153
N C D R D D P S C
3
4100



163
V Q D N C D Q H V
3
4101



174
L Q D L E D M S V
3
4102



216
I C G P E D V C E
3
4103



231
G Q D K V S Y E V
3
4104



243
H G D E E R F F V
3
4105



257
P D A G F T G L I
3
4106



275
S N E D F S A S P
3
4107



276
N E D F S A S P I
3
4108



297
I M T P S T L P P
3
4109



328
R K A G C K L T I
3
4110



336
I C P Q A E N R N
3
4111



338
P Q A E N R N D R
3
4112



354
L G Y V Q A P H K
3
4113



357
V Q A P H K T L P
3
4114



372
R N G E L Q D F P
3
4115



393
T R E P R D R S V
3
4116



408
G N L E V S P P V
3
4117



434
L P G S S G R R V
3
4118



436
G S S G R R V T Q
3
4119



452
A Q K V Q P P V E
3
4120



455
V Q P P V E L F V
3
4121



475
L S F V P A P D G
3
4122



500
F Q E K Q K C G H
3
4123



541
N K F V Q S C I D
3
4124



557
R E L G L A E C D
3
4125



573
F K T E R K K A T
3
4126



640
L H G E V H C G T
3
4127



644
V H C G T N V C R
3
4128



653
K P F S F K W W N
3
4129



7
V R V S L S H P T
2
4130



29
I Y G S V P E G T
2
4131



40
F E V Y G T P G V
2
4132



52
I S P N M E R G R
2
4133



58
R G R E R A D T R
2
4134



70
F D A T L E I I V
2
4135



90
S H V Q I S Y H S
2
4136



94
I S Y H S S H E P
2
4137



96
Y H S S H E P L P
2
4138



117
I S L D C D L N C
2
4139



137
R Q W V W G P S G
2
4140



154
C D R D D P S C D
2
4141



159
P S C D V Q D N C
2
4142



160
S C D V Q D N C D
2
4143



171
V H C L Q D L E D
2
4144



182
V M V L R T Q G P
2
4145



187
T Q G P A A L F D
2
4146



190
P A A L F D D H K
2
4147



197
H K L V L H T S S
2
4148



204
S S Y D A K R A Q
2
4149



214
F H I C G P E D V
2
4150



219
P E D V C E A Y R
2
4151



223
C E A Y R H V L G
2
4152



226
Y R H V L G Q D K
2
4153



235
V S Y E V P R L H
2
4154



265
I S F H V T L L D
2
4155



272
L D D S N E D F S
2
4156



282
S P I F T D T V V
2
4157



294
A P W I M T P S T
2
4158



299
T P S T L P P L E
2
4159



320
V D A V A E L A R
2
4160



324
A E L A R K A G C
2
4161



330
A G C K L T I C P
2
4162



332
C K L T I C P Q A
2
4163



342
N R N D R W I Q D
2
4164



355
G Y V Q A P H K T
2
4165



358
Q A P H K T L P V
2
4166



382
K R I L G P D F G
2
4167



385
L G P D F G Y V T
2
4168



390
G Y V T R E P R D
2
4169



394
R E P K D R S V S
2
4170



410
L F V S P P V V A
2
4171



413
S P P V V A N G K
2
4172



430
I G G N L P G S S
2
4173



431
G G N L P G S S G
2
4174



451
H A Q K V Q P P V
2
4175



479
P A P D G K G F R
2
4176



486
F R M L L A S P G
2
4177



487
R M L L A S P G A
2
4178



490
L A S P G A C F K
2
4179



497
F K L F Q F K Q K
2
4180



503
K Q K C G H G R A
2
4181



528
I N Q V L S N K D
2
4182



534
N K D L I N Y N K
2
4183



540
Y N K F V Q S C I
2
4184



570
P Q L F K T E R K
2
4185



581
T A F F P D L V N
2
4186



584
F P D L V N M L V
2
4187



593
L G K H L G I P K
2
4188



604
G P I I N G C C C
2
4189



609
G C C C L E E K V
2
4190



617
V R S L L E P L G
2
4191



636
P Y H M L H G E V
2
4192



638
H M L H G E V H C
2
4193



42
V Y G T P G V D I
1
4194



53
S P N M E R G R E
1
4195



55
N M E R G R E R A
1
4196



59
G R E R A D T R R
1
4197



60
R E R A D T R R W
1
4198



113
T C V D I S L D C
1
4199



121
C D L N C E G R Q
1
4200



127
G R Q D R N F V D
1
4201



152
V N C D R D D P S
1
4202



161
C D V Q D N C D Q
1
4203



203
T S S Y D A K R A
1
4204



227
R H V L G Q D K V
1
4205



236
S Y E V P R L H G
1
4206



239
V P R L H G D E E
1
4207



240
P R L H G D E E R
1
4208



281
A S P I F T D T V
1
4209



291
F R V A P W I M T
1
4210



300
P S T L P P L E V
1
4211



323
V A E L A R K A G
1
4212



327
A R K A G C K L T
1
4213



337
C P Q A E N R N D
1
4214



339
Q A E N R N D R W
1
4215



364
L P V V F D S P R
1
4216



370
S P R N G E L Q D
1
4217



397
R D R S V S G L D
1
4218



401
V S G L D S F G N
1
4219



404
L D S F G N L E V
1
4220



414
P P V V A N G K E
1
4221



435
P G S S G R R V T
1
4222



438
S G R R V T Q V V
1
4223



439
G R R V T Q V V R
1
4224



471
V D E F L S F V P
1
4225



493
P G A C F K L F Q
1
4226



514
F Q G V V D D E Q
1
4227



515
Q G V V D D E Q V
1
4228



544
V Q S C I D W N R
1
4229



545
Q S C I D W N R E
1
4230



548
I D W N R E V L K
1
4231



552
R E V L K R E L G
1
4232



569
I P Q L F K T E R
1
4233



577
R K K A T A F F P
1
4234



600
P K P F G P I I N
1
4235



603
F G P I I N G C C
1
4236



607
I N G C C C L E E
1
4237



642
G E V H C G T N V
1
4238



646
C G T N V C R K P
1
4239



648
T N V C R K P F S
1
4240







184P1E2 v.2: HLA Peptide


Scoring Results A26 9-mers SYFPEITHI












3
M T P S T L A P L
25
4241



6
S T L A P L E V Y
24
4242



7
T L A P L E V Y V
13
4243



1
W I M T P S T L A
10
4244



9
A P L E V Y V C R
7
4245



8
L A P L E V Y V C
5
4246



2
I M T P S T L A P
4
4247



4
T P S T L A P L E
2
4248



5
P S T L A P L E V
1
4249







184P1E2 v.3: HLA Peptide


Scoring Results A26 9-mers SYFPEITHI












2
E F L S F V P V P
18
4250



1
D E F L S F V P V
14
4251



7
V P V P D G K G F
13
4252



9
V P D G K G F R M
13
4253



6
F V P V P D G K G
12
4254



8
P V P D G K G F R
12
4255



3
F L S F V P V P D
9
4256



5
S F V P V P D G K
9
4257



4
L S F V P V P D G
7
4258





















TABLE XXVIII










SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.
















184P1E2 v.1: HLA Peptide


Scoring Results B 0702 9-mers SYFPEITHI












142
G P S G Y G G I L
25
4259



256
F P D A G F T G L
24
4260



480
A P D G K G F R M
22
4261



359
A P H K T L P V V
21
4262



294
A P W I M T P S T
20
4263



599
I P K P F G P I I
20
4264



101
E P L P L A Y A V
19
4265



478
V P A P D G K G F
19
4266



492
S P G A C F K L F
19
4267



622
E P L G L H C T F
19
4268



13
H P T S A V C V A
18
4269



282
S P I F T D T V V
18
4270



43
L P G S S G R R V
18
4271



584
F P D L V N M L V
17
4272



601
K P F G P I I N G
16
4273



3
L Q R I V R V S L
15
4274



97
H S S H E P L P L
15
4275



23
K V S T E V P R L
15
4276



395
E P R D R S V S G
15
4277



453
Q K V Q P P V E L
15
4278



104
P L A Y A V L Y L
14
4279



193
L F D D H K L V L
14
4280



246
E E R F F V E G L
14
4281



264
L I S F H V T L L
14
4282



326
L A R K A G C K L
14
4283



356
Y V Q A P H K T L
14
4284



370
S P R N G E L Q D
14
4285



423
Y P L G R I L I G
14
4286



467
A V G H V D E F L
14
4287



491
A S P G A C F K L
14
4288



504
Q K C G H G R A L
14
4289



505
K C G H G R A L L
14
4290



553
E V L K R E L G L
14
4291



579
K A T A F F P D L
14
4292



616
K V R S L L E P L
14
4293



33
V P E G T E M F E
13
4294



42
V T G T P G V D I
13
4295



79
V M N S P S N D L
13
4296



103
L P L A Y A V L Y
13
4297



158
D P S C D V Q D N
13
4298



177
L E D M S V M V L
13
4299



186
R T Q G P A A L F
13
4300



189
G P A A L F D D H
13
4301



222
V C E A Y R H V L
13
4302



239
V P R L H G D E E
13
4303



299
T P S T L P P L E
13
4304



368
F D S P R N G E L
13
4305



379
F P Y K R I L G P
13
4306



386
G P D F G T V T R
13
4307



417
V A N G K E Y P L
13
4308



421
K E Y P L G R I L
13
4309



457
P P V E L F V D W
13
4310



481
P D G K G F R M L
13
4311



482
D G K G F R M L L
13
4312



547
C I D W N R E V L
13
4313



585
P D L V N M L V L
13
4314



618
R S L L E P L G L
13
4315



632
D D F T P Y H M L
13
4316



18
V C V A G V E T L
12
4317



66
R R W R F D A T L
12
4318



82
S P S N D L N D S
12
4319



102
P L P L A Y A V L
12
4320



143
P S G Y G G I L L
12
4321



166
N C D Q H V H C L
12
4322



185
L R T Q G P A A L
12
4323



191
A A L F D D H K L
12
4324



208
A K R A Q V F H I
12
4325



253
G L S F P D A G F
12
4326



263
G L I S F H V T L
12
4327



298
M T P S T L P P L
12
4328



303
L P P L E V Y V C
12
4329



304
P P L E V Y V C R
12
4330



318
C F V D A V A E L
12
4331



328
R K A G C K L T I
12
4332



396
P R D R S V S G L
12
4333



410
L E V S P P V V A
12
4334



442
V T Q V V R D F L
12
4335



454
K V Q P P V E L F
12
4336



489
L L A S P G A C F
12
4337



524
K T I S I N Q V L
12
4338



569
I P Q L F K T E R
12
4339



598
G I P K P F G P I
12
4340



613
L E E K V R S L L
12
4341



10
S L E H P T S A V
11
4342



17
A V C V A G V E T
11
4343



45
T P G V D I Y I S
11
4344



53
S P N M E R G R E
11
4345



72
A T L E I I V V M
11
4346



95
S Y H S S H E P L
11
4347



126
E G R Q D R N F V
11
4348



179
D M S V M V L R T
11
4349



184
V L R T Q G P A A
11
4350



218
G P E D V C E A Y
11
4351



280
S A S P I F T D T
11
4352



300
P S T L P P L E V
11
4353



312
R V R N N T C F V
11
4354



337
C P Q A E N R N D
11
4355



346
R W I Q D E M E L
11
4356



364
L P V V F D S P R
11
4357



377
Q D F P Y K R I L
11
4358



402
S G L D S F G N L
11
4359



411
E V S P P V V A N
11
4360



426
G R I L I G G N L
11
4361



456
Q P P V E L F V D
11
4362



458
P V E L F V D W L
11
4363



564
C D I I D I P Q L
11
4364



575
T E R K K A T A F
11
4365



576
E R K K A T A F F
11
4366



582
A F F P D L V N M
11
4367



583
F F P D L V N M L
11
4368



589
N M L V L G K H L
11
4369



604
G P I I N G C C C
11
4370



612
C L E E K V R S L
11
4371



653
K P F S F K W W N
11
4372



19
C V A G V E T L V
10
4373



21
A G V E T L V D I
10
4374



57
E R G R E R A D T
10
4375



65
T R R W R F D A T
10
4376



98
S S H E P L P L A
10
4377



100
H E P L P L A Y A
10
4378



111
Y L T C V D I S L
10
4379



115
V D I S L D C D L
10
4380



144
S G Y G G I L L V
10
4381



155
D R D D P S C D V
10
4382



169
Q H V H C L Q D L
10
4383



195
D D H K L V L H T
10
4384



205
S Y D A K R A Q V
10
4385



248
R F F V E G L S F
10
4386



257
P D A G F T G L I
10
4387



278
D F S A S P I F T
10
4388



283
P I F T D T V V F
10
4389



291
F R V A P W I M T
10
4390



295
P W I M T P S T L
10
4391



314
R N N T C F V D A
10
4392



358
Q A P H K T L P V
10
4393



376
L Q D F P Y K R I
10
4394



381
Y K R I L G P D F
10
4395



384
I L G P D F G Y V
10
4396



404
L D S F G N L E V
10
4397



413
S P P V V A N G K
10
4398



414
P P V V A N G K E
10
4399



435
P G S S G R R V T
10
4400



438
S G R R V T Q V V
10
4401



455
V Q P P V E L F V
10
4402



460
E L F V D W L A V
10
4403



472
D E F L S F V P A
10
4404



506
C G H G R A L L F
10
4405



529
N Q V L S N K D L
10
4406



551
N R E V L K R E L
10
4407



591
L V L C K H L G I
10
4408



595
K H L G I P K P F
10
4409



605
P I I N G C C C L
10
4410



635
T P Y H M L H G E
10
4411



12
E H P T S A V C V
9
4412



15
T S A V C V A G V
9
4413



31
G S V P E G T E M
9
4414



62
R A D T R R W R F
9
4415



6
D T R R W R F D A
9
4416



68
W R F D A T L E I
9
4417



69
R F D A T L E I I
9
4418



206
T D A K R A Q V F
9
4419



241
R L H G D E E R F
9
4420



243
H G D S E R F F V
9
4421



251
V E G L S F P D A
9
4422



281
A S P I F T D T V
9
4423



302
T L P P L E V Y V
9
4424



322
A V A E L A R K A
9
4425



327
A R K A G C K L T
9
4426



351
E M E L G Y V Q A
9
4427



360
P H K T L P V V F
9
4428



420
G K E Y P L G R I
9
4429



437
S S G R R V T Q V
9
4430



444
Q V V R D F L H A
9
4431



451
H A Q K V Q P P V
9
4432



466
L A V G H V D E F
9
4433



469
G H V D E F L S F
9
4434



483
G K G F R M L L A
9
4435



517
V V D D E Q V K T
9
4436



580
A T A F F P D L V
9
4437



623
P L G L H C T F I
9
4438



1
M S L Q R I V R V
8
4439



29
I Y G S V P E G T
8
4440



37
T E M F E V Y G T
8
4441



40
F E V T G T P G V
8
4442



44
G T P G V D I Y I
8
4443



86
D L N D S H V Q I
8
4444



109
V L Y L T C V D I
8
4445



176
D L E D M S V M V
8
4446



192
A L F D D H K L V
8
4447



203
T S S Y D A K R A
8
4448



231
G Q D K V S Y E V
8
4449



242
L H G D E E R F F
8
4450



254
L S F P D A G F T
8
4451



259
A G F T G L I S F
8
4452



262
T G L I S F H V T
8
4453



273
D D S N E D F S A
8
4454



276
N E D F S A S P I
8
4455



277
E D F S A S P I F
8
4456



285
F T D T V V F R V
8
4457



286
T D T V V F R V A
8
4458



290
V F R V A P W I M
8
4459



315
N N T C F V D A V
8
4460



316
N T C F V D A V A
8
4461



319
F V D A V A E L A
8
4462



340
A E N R N D R W I
8
4463



344
N D R W I Q D E M
8
4464



371
P R N G E L Q D F
8
4465



385
L G P D F G Y V T
8
4466



408
G N L E V S P P V
8
4467



409
N L E V S P P V V
8
4468



422
E Y P L G R I L I
8
4469



441
R V T Q V V R D F
8
4470



470
H V D E F L S F V
8
4471



487
R M L L A S P G A
8
4472



509
G R A L L F Q G V
8
4473



510
R A L L F Q G V V
8
4474



518
V D D E Q V K T I
8
4475



558
E L G L A E C D I
8
4476



566
I I D I P Q L F K
8
4477



567
I D I P Q L F K T
8
4478



574
K T E R K K A T A
8
4479



631
I D D F T P Y H M
8
4480



642
G E V H C G T N V
8
4481



647
G T N V C R K P F
8
4482



650
V C R K P F S F R
8
4483



654
P F S F K W W N M
8
4484



7
V R V S L E H P T
7
4485



9
V S L E H P T S A
7
4486



32
S V P E G T E M F
7
4487



34
P E G T E M F E V
7
4488



48
V D I Y I S P N M
7
4489



55
N M E R G R E R A
7
4490



70
F D A T L E I I V
7
4491



71
D A T L E I I V V
7
4492



84
S N D L N D S H V
7
4493



105
L A Y A V L Y L T
7
4494



107
Y A V L Y L T C V
7
4495



125
C E G R Q D R N F
7
4496



132
N F V D K R Q W V
7
4497



163
V Q D N C D Q H V
7
4498



174
L Q D L E D M S V
7
4499



175
Q D L E D M S V M
7
4500



183
M V L R T Q G P A
7
4501



217
C G P E D V C E A
7
4502



227
R H V L G Q D K V
7
4503



261
F T G L I S F H V
7
4504



271
L L D D S N E D F
7
4505



289
V V F R V A P W I
7
4506



305
P L E V Y V C R V
7
4507



309
Y V C R V R N N T
7
4508



311
C R V R N N T C F
7
4509



332
C K L T I C P Q A
7
4510



361
H K T L P V V F D
7
4511



393
T R E P R D R S V
7
4512



399
R S V S G L D S F
7
4513



436
G S S G R R V T Q
7
4514



447
R D F L H A Q K V
7
4515



463
V D W L A V G H V
7
4516



503
K Q K C G H G R A
7
4517



520
D E Q V K T I S I
7
4518



535
K D L I N Y N K F
7
4519



536
D L I N Y N K F V
7
4520



559
L G L A E C D I I
7
4521



572
L F K T E R K K A
7
4522



573
F K T E R K K A T
7
4523



627
H C T F I D D F T
7
4524



640
L H G E V H C G T
7
4525



655
F S F K W W N M V
7
4526



14
P T S A V C V A G
6
4527



25
T L V D I Y G S V
6
4528



141
W G P S G Y G G I
6
4529



145
G Y G G I L L V N
6
4530



172
H C L Q D L E D M
6
4531



200
V L H T S S Y D A
6
4532



214
F H I C G P E D V
6
4533



215
H I C G P E D V C
6
4534



221
D V C E A Y R H V
6
4535



236
S Y S V P R L H G
6
4536



297
I M T P S T L P P
6
4537



349
Q D E M S L G Y V
6
4538



355
G Y V Q A P H K T
6
4539



383
R I L G P D F G Y
6
4540



392
V T R E P R D R S
6
4541



428
I L I G G N L P G
6
4542



459
V E L F V D W L A
6
4543



511
A L L F Q G V V D
6
4544



515
Q G V V D D E Q V
6
4545



523
V K T I S I N Q V
6
4546



530
Q V L S N K D L I
6
4547



540
Y N K F V Q S C I
6
4548



546
S C I D W N R E V
6
4549



554
V L K R E L G L A
6
4550



555
L K R E L G L A E
6
4551



561
L A E C D I I D I
6
4552



565
D I I D I P Q L F
6
4553



581
T A F F P D L V N
6
4554



596
H L G I P K P F G
6
4555



609
G C C C L E E K V
6
4556



621
L E P L G L H C T
6
4557



626
L H C T F I D D F
6
4558



636
P Y H M L H G E V
6
4559



649
N V C R K P F S F
6
4560



20
V A G V E T L V D
5
4561



30
Y G S V P E G T E
5
4562



56
M E R G R E R A D
5
4563



60
R E R A D T R R W
5
4564



106
A Y A V L Y L T C
5
4565



128
R Q D R N F V D K
5
4566



139
W V W G P S G Y G
5
4567



187
T Q G P A A L F D
5
4568



216
I C G P E D V C E
5
4569



284
I F T D T V V F R
5
4570



296
W I M T P S T L P
5
4571



301
S T L P P L E V Y
5
4572



367
V F D S P R N G E
5
4573



398
D R S V S G L D S
5
4574



400
S V S G L D S F G
5
4575



418
A N G K E Y P L G
5
4576



439
G R R V T Q V V R
5
4577



8
R V S L E H P T S
4
4578



28
D I Y G S V P E G
4
4579



41
E V Y G T P G V D
4
4580



50
I Y I S P N M E R
4
4581



67
R W R F D A T L E
4
4582



99
S H E P L P L A Y
4
4583



108
A V L T L T C V D
4
4584



156
R D D P S C D V Q
4
4585



178
E D M S V M V L R
4
4586



224
E A Y R H V L G Q
4
4587



225
A Y R H V L G Q D
4
4588



258
D A G F T G L I S
4
4589



265
I S F H V T L L D
4
4590



279
F S A S P I F T D
4
4591



287
D T V V F R V A P
4
4592



307
E V Y V C R V R N
4
4593



320
V D A V A E L A R
4
4594



324
A E L A R K A G C
4
4595



330
A G C K L T I C P
4
4596



334
L T I C P Q A E N
4
4597



341
E N R N D R W I Q
4
4598



348
I Q D E M E L G Y
4
4599



352
M E L G Y V Q A P
4
4600



353
E L G Y V Q A P H
4
4601



387
P D F G Y V T R E
4
4602



429
L I G G N L P G S
4
4603



431
G G N L P G S S G
4
4604



440
R R V T Q V V R D
4
4605



452
A Q K V Q P P V E
4
4606



473
E F L S F V P A P
4
4607



474
F L S F V P A P D
4
4608



475
L S F V P A P D G
4
4609



477
F V P A P D G K G
4
4610



484
K G F R M L L A S
4
4611



490
L A S P G A C F K
4
4612



493
P G A C F K L F Q
4
4613



495
A C F K L F Q E K
4
4614



507
G H G R A L L F Q
4
4615



525
T I S I N Q V L S
4
4616



526
I S I N Q V L S N
4
4617



531
V L S N K D L I N
4
4618



538
I N Y N K F V Q S
4
4619



578
K K A T A F F P D
4
4620



607
I N G C C C L E E
4
4621



620
L L E P L G L H C
4
4622



644
V H C G T N V C R
4
4623



5
R I V R V S L E H
3
4624



6
I V R V S L E H P
3
4625



11
L E H P T S A V C
3
4626



26
L V D I Y G S V P
3
4627



27
V D I Y G S V P E
3
4628



35
E G T E M F E V Y
3
4629



39
M F E V Y G T P G
3
4630



51
Y I S P N M E R G
3
4631



58
R G R E R A D T R
3
4632



61
E R A D T R R W R
3
4633



63
A D T R R W R F D
3
4634



73
T L E I I V V M N
3
4635



80
M N S P S N D L N
3
4636



96
Y H S S H E P L P
3
4637



117
I S L D C D L N C
3
4638



129
Q D R N F V D K R
3
4639



133
F V D K R Q W V W
3
4640



135
D K R Q W V W G P
3
4641



137
R Q W V W G P S G
3
4642



140
V W G P S G Y G G
3
4643



146
Y G G I L L V N C
3
4644



181
S V M V L R T Q G
3
4645



194
F D D H K L V L H
3
4646



202
H T S S Y D A K R
3
4647



207
D A K R A Q V F H
3
4648



209
K R A Q V F H I C
3
4649



223
C E A Y R H V L G
3
4650



228
H V L G Q D K V S
3
4651



229
V L G Q D K V S Y
3
4652



233
D K V S Y E V P R
3
4653



245
D E E R F F V E G
3
4654



250
F V E G L S F P D
3
4655



260
G F T G L I S F H
3
4656



288
T V V F R V A P W
3
4657



292
R V A P W I M T P
3
4658



317
T C F V D A V A E
3
4659



329
K A G C K L T I C
3
4660



350
D E M E L G Y V Q
3
4661



357
V Q A P H K T L P
3
4662



362
K T L P V V F D S
3
4663



372
R N G E L Q D F P
3
4664



394
R E P R D R S V S
3
4665



397
R D R S V S G L D
3
4666



405
D S F G N L E V S
3
4667



406
S F G N L E V S P
3
4668



412
V S P P V V A N G
3
4669



445
V V R D F L H A Q
3
4670



449
F L H A Q K V Q P
3
4671



450
L H A Q K V Q P P
3
4672



462
F V D W L A V G H
3
4673



464
D W L A V G H V D
3
4674



465
W L A V G H V D E
3
4675



485
G F R M L L A S P
3
4676



502
E K Q K C G H G R
3
4677



508
H G R A L L F Q G
3
4678



512
L L F Q G V V D D
3
4679



513
L F Q G V V D D E
3
4680



548
I D W N R E V L K
3
4681



549
D W N R E V L K R
3
4682



557
R E L G L A E C D
3
4683



563
E C D I I D I P Q
3
4684



577
R K K A T A F F P
3
4685



587
L V N M L V L G K
3
4686



606
I I N G C C C L E
3
4687



614
E E K V R S L L E
3
4688



615
E K V R S L L E P
3
4689



617
V R S L L E P L G
3
4690



619
S L L E P L G L H
3
4691



629
T F I D D F T P Y
3
4692



630
F I D D F T P Y H
3
4693



637
Y H M L H G E V H
3
4694



639
M L H G E V H C G
3
4695



643
E V H C G T N V C
3
4696



2
S L Q R I V R V S
2
4697



36
G T E M F E V Y G
2
4698



43
Y G T P G V D I Y
2
4699



47
G V D I Y I S P N
2
4700



54
P N M E R G R E R
2
4701



76
I I V V M N S P S
2
4702



77
I V V M N S P S N
2
4703



81
N S P S N D L N D
2
4704



87
L N D S H V Q I S
2
4705



88
N D S H V Q I S Y
2
4706



93
Q I S Y H S S H E
2
4707



113
T C V D I S L D C
2
4708



116
D I S L D C D L N
2
4709



124
N C E G R Q D R N
2
4710



134
V D K R Q W V W G
2
4711



136
K R Q W V W G P S
2
4712



153
N C D R D D P S C
2
4713



154
C D R D D P S C D
2
4714



157
D D P S C D V Q D
2
4715



165
D N C D Q H V H C
2
4716



168
D Q H V H C L Q D
2
4717



171
V H C L Q D L E D
2
4718



198
K L V L H T S S Y
2
4719



211
A Q V F H I C G P
2
4720



213
V F H I C G P E D
2
4721



219
P E D V C E A Y R
2
4722



244
G D E E R F F V E
2
4723



252
E G L S F P D A G
2
4724



266
S F H V T L L D D
2
4725



272
L D D S N E D F S
2
4726



274
D S N E D F S A S
2
4727



293
V A P W I M T P S
2
4728



310
V C R V R N N T C
2
4729



321
D A V A E L A R K
2
4730



323
V A E L A R K A G
2
4731



325
E L A R K A G C K
2
4732



333
K L T I C P Q A E
2
4733



338
P Q A E N R N D R
2
4734



343
R N D R W I Q D E
2
4735



374
G E L Q D F P Y K
2
4736



382
K R I L G P D F G
2
4737



388
D F G Y V T R E P
2
4738



389
F G Y V T R E P R
2
4739



403
G L D S F G N L E
2
4740



407
F G N L E V S P P
2
4741



415
P V V A N G K E Y
2
4742



419
N G K E Y P L G R
2
4743



425
L G R I L I G G N
2
4744



427
R I L I G G N L P
2
4745



430
I G G N L P G S S
2
4746



433
N L P G S S G R R
2
4747



446
V R D F L H A Q K
2
4748



461
L F V D W L A V G
2
4749



486
F R M L L A S P G
2
4750



488
M L L A S P G A C
2
4751



521
E Q V K T I S I N
2
4752



539
N Y N K F V Q S C
2
4753



544
V Q S C I D W N R
2
4754



550
W N R E V L K R E
2
4755



552
R E V L K R E L G
2
4756



556
K R E L G L A E C
2
4757



562
A E C D I I D I P
2
4758



586
D L V N M L V L G
2
4759



592
V L G K H L G I P
2
4760



593
L G K H L G I P K
2
4761



602
P F G P I I N G C
2
4762



611
C C L E E K V R S
2
4763



634
F T P Y H M L H G
2
4764



638
H M L H G E V H C
2
4765



645
H C G T N V C R K
2
4766



648
T N V C R K P F S
2
4767



652
R K P F S F K W W
2
4768



4
Q R I V R V S L E
1
4769



16
S A V C V A G V E
1
4770



22
G V E T L V D I Y
1
4771



24
E T L V D I Y G S
1
4772



38
E M F E V Y G T P
1
4773



52
I S P N M E R G R
1
4774



59
G R E R A D T R R
1
4775



74
L E I I V V M N S
1
4776



75
E I I V V M N S P
1
4777



78
V V M N S P S N D
1
4778



85
N D L N D S H V Q
1
4779



9
I S Y H S S H E P
1
4780



110
L Y L T C V D I S
1
4781



114
C V D I S L D C D
1
4782



118
S L D C D L N C E
1
4783



120
D C D L N C E G R
1
4784



122
D L N C E G R Q D
1
4785



123
L N C E G R Q D R
1
4786



127
G R Q D R N F V D
1
4787



131
R N F V D K R Q W
1
4788



147
G G I L L V N C D
1
4789



149
I L L V N C D R D
1
4790



151
L V N C D R D D P
1
4791



152
V N C D R D D P S
1
4792



160
S C D V Q D N C D
1
4793



164
Q D N C D Q H V H
1
4794



182
V M V L R T Q G P
1
4795



188
Q G P A A L F D D
1
4796



190
P A A L F D D H K
1
4797



197
H K L V L H T S S
1
4798



201
L H T S S Y D A K
1
4799



204
S S Y D A K R A Q
1
4800



210
R A Q V F H I C G
1
4801



212
Q V F H I C G P E
1
4802



220
E D V C E A Y R H
1
4803



226
Y R H V L G Q D K
1
4804



230
L G Q D K V S Y E
1
4805



232
Q D K V S Y E V P
1
4806



237
Y E V P R L H G D
1
4807



238
E V P R L H C D E
1
4808



247
E R F F V E G L S
1
4809



249
F F V E G L S F P
1
4810



255
S F P D A G F T G
1
4811



275
S N E D F S A S P
1
4812



306
L E V Y V C R V R
1
4813



308
V Y V C R V R N N
1
4814



313
V R N N T C F V D
1
4815



331
G C K L T I C P Q
1
4816



335
T I C P Q A E N R
1
4817



336
I C P Q A E N R N
1
4818



354
L G Y V Q A P H K
1
4819



363
T L P V V F D S P
1
4820



365
P V V F D S P R N
1
4821



369
D S P R N G E L Q
1
4822



373
N G E L Q D F P Y
1
4823



375
E L Q D F P Y K R
1
4824



378
D F P Y K R I L G
1
4825



380
P Y K R I L G P D
1
4826



390
G Y V T R E P R D
1
4827



401
V S G L D S F G N
1
4828



416
V V A N G K E Y P
1
4829



424
P L G R I L I G G
1
4830



432
G N L P G S S G R
1
4831



448
D F L H A Q K V Q
1
4832



471
V D E F L S F V P
1
4833



476
S F V P A P D G K
1
4834



479
P A P D G K G F R
1
4835



498
K L F Q E K Q K C
1
4836



500
F Q E K Q K C G H
1
4837



514
F Q G V V D D E Q
1
4838



516
G V V D D E Q V K
1
4839



519
D D E Q V K T I S
1
4840



528
I N Q V L S N K D
1
4841



532
L S N K D L I N Y
1
4842



534
N K D L I N Y N K
1
4843



537
L I N Y N K F V Q
1
4844



542
K F V Q S C I D W
1
4845



560
G L A E C D I I D
1
4846



568
D I P Q L F K T E
1
4847



570
P Q L F K T E R K
1
4848



571
Q L F K T E R K K
1
4849



588
V N M L V L G K H
1
4850



594
G K H L G I P K P
1
4851



597
L G I P K P F G P
1
4852



610
C C C L E E K V R
1
4853



624
L G L H C T F I D
1
4854



625
G L H C T F I D D
1
4855



633
D F T P Y H M L H
1
4856



651
C R K P F S F K W
1
4857



656
S F K W W N M V P
1
4858







184P1E2 v.2: HLA Peptide


Scoring Results B 0702 9-mers SYFPEITHI












9
A P L E V Y V C R
14
4859



4
T P S T L A P L E
13
4860



3
M T P S T L A P L
12
4861



1
W I M T P S T L A
11
4862



7
T L A P L E V Y V
10
4863



5
P S T L A P L E V
8
4864



2
I M T P S T L A P
6
4865



6
S T L A P L E V Y
5
4866



8
L A P L E V Y V C
2
4867







184P1E2 v.3: HLA Peptide


Scoring Results B 0702 9-mers SYFPEITHI












9
V P D G K G F R M
20
4868



7
V P V P D G K G F
18
4869



1
D E F L S F V P V
10
4870



3
F L S F V P V P D
5
4871



4
L S F V P V P D G
5
4872



2
E F L S F V P V P
4
4873



6
F V P V P D G K G
3
4874



5
S F V P V P D G K
1
4875



8
P V P D G K G F R
1
4876

















TABLE XXIX







184P1E2 v.1: HLA Peptide


Scoring Results B08 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







612
C L E E K V R S L
29
4877



417
V A N G K E Y P L
25
4878



395
E P R D R S V S G
23
4879



575
T E R K K A T A F
23
4880



599
I P K P F G P I I
23
4881



368
F D S P R N G E L
21
4882



614
E E K V R S L L E
21
4883



326
L A R K A G C K L
20
4884



482
D G K G F R M L L
20
4885



501
Q E K Q K C G H G
20
4886



553
E V L K R E L G L
20
4887



246
E E R F F V E G L
19
4888



62
R A D T R R W R F
18
4889



104
P L A Y A V L Y L
18
4890



263
G L I S F H V T L
18
4891



325
E L A R K A G C K
18
4892



576
E R K K A T A F F
18
4893



3
L Q R I V R V S L
17
4894



102
P L P L A Y A V L
17
4895



142
G P S G Y G C I L
17
4896



256
F P D A G F T G L
17
4897



310
V C R V R N N T C
17
4898



360
P H K T L P V V F
17
4899



520
D E Q V K T I S I
17
4900



531
V L S N K D L I N
17
4901



554
V L K R E L G L A
17
4902



58
R G R E R A D T R
16
4903



111
Y L T C V D I S L
16
4904



379
F P Y K R I L C P
16
4905



480
A P D G K G F R M
16
4906



540
Y N K F V Q S C I
16
4907



591
L V L G K H L G I
16
4908



616
K V R S L L E P L
16
4909



649
N V C R K P F S F
16
4910



56
M E R G R E R A D
15
4911



65
T R R W R F D A T
15
4912



109
V L Y L T C V D I
15
4913



207
D A K R A Q V F H
15
4914



239
V P R L H G D E E
15
4915



264
L I S F H V T L L
15
4916



423
Y P L G R I L I G
15
4917



492
S P G A C F K L F
15
4918



494
G A C F K L F Q E
15
4919



558
E L G L A E C D I
15
4920



184
V L R T Q G P A A
14
4921



191
A A L F D D H K L
14
4922



206
Y D A K R A Q V F
14
4923



329
K A G C K L T I C
14
4924



358
Q A P H K T L P V
14
4925



370
S P R N G E L Q D
14
4926



478
V P A P D G K G F
14
4927



506
C G H G R A L L F
14
4928



547
C I D W N R E V L
14
4929



579
K A T A F F P D L
14
4930



605
P I I N G C C C L
14
4931



622
E P L G L H C T F
14
4932



86
D L N D S H V Q I
13
4933



208
A K R A Q V F H I
13
4934



253
G L S F P D A G F
13
4935



271
L L D D S N E D F
13
4936



331
G C K L T I C P Q
13
4937



339
Q A E N R N D R W
13
4938



499
L F Q E K Q K C G
13
4939



529
N Q V L S N K D L
13
4940



533
S N K D L I N Y N
13
4941



573
F K T E R K K A T
13
4942



623
P L G L H C T F I
13
4943



656
S F K W W N M V P
13
4944



18
V C V A G V E T L
12
4945



95
S Y H S S H E P L
12
4946



134
V D K R Q W V W G
12
4947



194
F D D H K L V L H
12
4948



205
S Y D A K R A Q V
12
4949



234
K V S Y E V P R L
12
4950



241
R L H G D E E R F
12
4951



318
C F V D A V A E L
12
4952



381
Y K R I L G P D F
12
4953



402
S G L D S F G N L
12
4954



419
N G K E Y P L G R
12
4955



426
G R I L I G G N L
12
4956



442
V T Q V V R D F L
12
4957



453
Q K V Q P P V E L
12
4958



458
P V E L F V D W L
12
4959



466
L A V G H V D E F
12
4960



489
L L A S P G A C F
12
4961



538
I N Y N K F V Q S
12
4962



551
N R E V L K R E L
12
4963



583
F F P D L V N M L
12
4964



589
N M L V L G K H L
12
4965



54
P N M E R G R E R
11
4966



79
V M N S P S N D L
11
4967



97
H S S H E P L P L
11
4968



115
V D I S L D C D L
11
4969



143
P S G Y G C I L L
11
4970



166
N C D Q H V H C L
11
4971



177
L E D M S V M V L
11
4972



185
L R T Q G P A A L
11
4973



196
D H K L V L H T S
11
4974



222
V C E A Y R H V L
11
4975



230
L G Q D K V S Y E
11
4976



232
Q D K V S Y E V P
11
4977



244
C D E E R F F V E
11
4978



283
P I F T D T V V F
11
4979



327
A R K A G C K L T
11
4980



346
R W I Q D E M E L
11
4981



356
Y V Q A P H K T L
11
4982



378
D F P Y K R I L G
11
4983



396
P R D R S V S C L
11
4984



437
S S G R R V T Q V
11
4985



450
L H A Q K V Q P P
11
4986



452
A Q K V Q P P V E
11
4987



496
C F K L F Q E K Q
11
4988



504
Q K C G H G R A L
11
4989



518
V D D S Q V K T I
11
4990



52
K T I S I N Q V L
11
4991



552
R E V L K R E L G
11
4992



561
L A E C D I I D I
11
4993



564
C D I I D I P Q L
11
4994



572
L F K T E R K K A
11
4995



585
P D L V N M L V L
11
4996



598
G I P K P F G P I
11
4997



618
R S L L E P L G L
11
4998



1
M S L Q R I V R V
10
4999



4
Q R I V R V S L E
10
5000



32
S V P E G T E M F
10
5001



66
R R W R F D A T L
10
5002



127
G R Q D R N F V D
10
5003



132
N F V D K R Q W V
10
5004



169
Q H V H C L Q D L
10
5005



182
V M V L R T Q G P
10
5006



193
L F D D H K L V L
10
5007



223
C E A Y R H V L G
10
5008



295
P W I M T P S T L
10
5009



298
M T P S T L P P L
10
5010



308
V Y V C R V R N N
10
5011



377
Q D F P Y K R I L
10
5012



380
P Y K R I L G P D
10
5013



421
K E Y P L G R I L
10
5014



436
G S S G R R V T Q
10
5015



460
E L F V D W L A V
10
5016



467
A V G H V D E F L
10
5017



481
P D G K G F R M L
10
5018



491
A S P C A C F K L
10
5019



503
K Q K C G H G R A
10
5020



505
K C G H G R A L L
10
5021



522
Q V K T I S I N Q
10
5022



548
I D W N R S V L K
10
5023



565
D I I D I P Q L F
10
5024



570
P Q L F K T E R K
10
5025



574
K T E R K K A T A
10
5026



577
R K K A T A F F P
10
5027



593
L G K H L G I P K
10
5028



597
L C I P K P F G P
10
5029



613
L E E K V R S L L
10
5030



619
S L L E P L G L H
10
5031



632
D D F T P Y H M L
10
5032



651
C R K P F S F K W
10
5033



654
P F S F K W W N N
10
5034



2
S L Q R I V R V S
9
5035



10
S L E H P T S A V
9
5036



21
A G V E T L V D I
9
5037



101
E P L P L A Y A V
9
5038



133
F V D K R Q W V W
9
5039



152
V N C D R D D P S
9
5040



288
T V V F R V A P W
9
5041



304
P P L E V Y V C R
9
5042



353
E L G Y V Q A P H
9
5043



390
G Y V T R E P R D
9
5044



392
V T R E P R D R S
9
5045



413
S P P V V A N G K
9
5046



422
E Y P L G R I L I
9
5047



439
G R R V T Q V V R
9
5048



443
T Q V V R D F L H
9
5049



457
P P V E L F V D W
9
5050



483
G K G F R M L L A
9
5051



550
W N R E V L K R E
9
5052



555
L K R E L G L A E
9
5053



560
G L A S C D I I D
9
5054



569
I P Q L F K T E R
9
5055



58
F P D L V N M L V
9
5056



647
G T N V C R K P F
9
5057



6
I V R V S L E H P
8
5058



13
H P T S A V C V A
8
5059



42
V Y G T P G V D I
8
5060



45
T P G V D I Y I S
8
5061



53
S P N M E R G R E
8
5062



63
A D T R R W R F D
8
5063



73
T L E I I V V M N
8
5064



82
S P S N D L N D S
8
5065



118
S L D C D L N C E
8
5066



124
N C E G R Q D R N
8
5067



126
E G R Q D R N F V
8
5068



149
I L L V N C D R D
8
5069



218
C P E D V C E A Y
8
5070



229
V L G Q D K V S Y
8
5071



237
Y E V P R L H G D
8
5072



277
E D F S A S P I F
8
5073



282
S P I F T D T V V
8
5074



290
V F R V A P W I M
8
5075



303
L P P L E V Y V C
8
5076



305
P L E V Y V C R V
8
5077



323
V A E L A R K A G
8
5078



324
A E L A R K A G C
8
5079



341
E N R N D R W I Q
8
5080



342
N R N D R W I Q D
8
5081



375
E L Q D F P Y K R
8
5082



386
G P D F G Y V T R
8
5083



393
T R E P R D R S V
8
5084



414
P P V V A N G K E
8
5085



420
G K E Y P L G R I
8
5086



428
I L I G G N L P G
8
5087



438
S G R R V T Q V V
8
5088



445
V V R D F L H A Q
8
5089



449
F L H A Q K V Q P
8
5090



465
W L A V G H V D E
8
5091



512
L L F Q G V V D D
8
5092



536
D L I N Y N K F V
8
5093



648
T N V C R K P F S
8
5094



33
V P E G T E M F E
7
5095



44
G T P G V D I Y I
7
5096



68
W R F D A T L S I
7
5097



75
E I I V V M N S P
7
5098



76
I I V V M N S P S
7
5099



103
L P L A Y A V L Y
7
5100



129
Q D R N F V D K R
7
5101



135
D K R Q W V W G P
7
5102



148
G I L L V N C D R
7
5103



158
D P S C D V Q D N
7
5104



176
D L E D M S V M V
7
5105



189
G P A A L F D D H
7
5106



192
A L F D D H K L V
7
5107



198
K L V L H T S S Y
7
5108



200
V L H T S S Y D A
7
5109



224
E A Y R H V L G Q
7
5110



248
R F F V E G L S F
7
5111



270
T L L D D S N E D
7
5112



289
V V F R V A P W I
7
5113



302
T L P P L E V Y V
7
5114



311
C R V R N N T C F
7
5115



333
K L T I C P Q A E
7
5116



337
C P Q A E N R N D
7
5117



34
N D R W I Q D E M
7
5118



363
T L P V V F D S P
7
5119



364
L P V V F D S P R
7
5120



376
L Q D F P Y K R I
7
5121



384
I L G P D F G Y V
7
5122



403
G L D S F G N L E
7
5123



409
N L E V S P P V V
7
5124



425
L G R I L I G G N
7
5125



441
R V T Q V V R D F
7
5126



45
K V Q P P V E L F
7
5127



456
Q P P V E L F V D
7
5128



469
G H V D E F L S F
7
5129



47
F L S F V P A P D
7
5130



485
G F R M L L A S P
7
5131



488
M L L A S P G A C
7
5132



508
H G R A L L F Q G
7
5133



511
A L L F Q G V V D
7
5134



586
D L V N M L V L G
7
5135



590
M L V L G K H L G
7
5136



592
V L G K H L G I P
7
5137



595
K H L G I P K P F
7
5138



596
H L G I P K P F G
7
5139



601
K P F G P I I N G
7
5140



604
G P I I N C C C C
7
5141



625
G L H C T F I D D
7
5142



626
L H C T F I D D F
7
5143



639
M L H G E V H C G
7
5144



650
V C R K P F S F K
7
5145



16
S A V C V A G V E
6
5146



20
V A G V E T L V D
6
5147



25
T L V D I Y C S V
6
5148



28
D I Y G S V P B G
6
5149



60
R E R A D T R R W
6
5150



64
D T R R W R F D A
6
5151



67
R W R F D A T L E
6
5152



69
R F D A T L E I I
6
5153



71
D A T L E I I V V
6
5154



122
D L N C E G R Q D
6
5155



125
C E G R Q D R N F
6
5156



141
W G P S G Y G C I
6
5157



150
L L V N C D R D D
6
5158



154
C D R D D P S C D
6
5159



173
C L Q D L E D M S
6
5160



186
R T Q C P A A L F
6
5161



225
A Y R H V L G Q D
6
5162



242
L H G D E E R F F
6
5163



257
P D A G F T C L I
6
5164



259
A G F T G L I S F
6
5165



276
N E D F S A S P I
6
5166



280
S A S P I F T D T
6
5167



294
A P W I M T P S T
6
5168



299
T P S T L P P L E
6
5169



312
R V R N N T C F V
6
5170



328
R K A C C K L T I
6
5171



340
A E N R N D R W I
6
5172



359
A P H K T L P V V
6
5173



371
P R N G E L Q D F
6
5174



397
R D R S V S G L D
6
5175



399
R S V S G L D S F
6
5176



424
P L G R I L I G G
6
5177



433
N L P G S S G R R
6
5178



43
L P G S S G R R V
6
5179



498
K L F Q E K Q K C
6
5180



527
S I N Q V L S N K
6
5181



530
Q V L S N K D L I
6
5182



535
K D L I N Y N K F
6
5183



559
L G L A E C D I I
6
5184



571
Q L F K T E R K K
6
5185



620
L L E P L C L H C
6
5186



635
T P Y H M L H G E
6
5187



653
K P F S F K W W N
6
5188



5
R I V R V S L E H
5
5189



93
Q I S Y H S S H E
5
5190



107
Y A V L Y L T C V
5
5191



116
D I S L D C D L N
5
5192



190
P A A L F D D H K
5
5193



210
R A Q V F H I C G
5
5194



215
H I C G P E D V C
5
5195



293
V A P W I M T P S
5
5196



411
B V S P P V V A N
5
5197



451
H A Q K V Q P P V
5
5198



510
R A L L F Q G V V
5
5199



525
T I S I N Q V L S
5
5200



537
L I N Y N K F V Q
5
5201



566
I I D I P Q L F K
5
5202



568
D I P Q L F K T E
5
5203



606
I I N C C C C L E
5
5204



630
F I D D F T P Y H
5
5205



24
E T L V D I Y G S
4
5206



35
E G T E M F E V Y
4
5207



38
E M F E V Y G T P
4
5208



49
D I Y I S P N M E
4
5209



51
Y I S P N M B R G
4
5210



72
A T L E I I V V M
4
5211



98
S S H E P L P L A
4
5212



105
L A Y A V L Y L T
4
5213



217
C C P E D V C E A
4
5214



258
D A G F T G L I S
4
5215



296
W I M T P S T L P
4
5216



307
E V Y V C R V R N
4
5217



321
D A V A E L A R K
4
5218



335
T I C P Q A E N R
4
5219



347
W I Q D E M E L G
4
5220



351
E M E L G Y V Q A
4
5221



361
H K T L P V V F D
4
5222



383
R I L G P D F G Y
4
5223



427
R I L I G G N L P
4
5224



429
L I G G N L P G S
4
5225



473
E F L S F V P A P
4
5226



479
P A P D G K G F R
4
5227



490
L A S P G A C F K
4
5228



546
S C I D W N R E V
4
5229



581
T A F F P D L V N
4
5230



9
V S L E H P T S A
3
5231



12
E H P T S A V C V
3
5232



17
A V C V A G V F T
3
5233



27
V D I Y G S V P E
3
5234



50
I Y I S P N M E R
3
5235



84
S N D L N D S H V
3
5236



99
S H E P L P L A Y
3
5237



144
S G Y G G I L L V
3
5238



147
G G I L L V N C D
3
5239



160
S C D V Q D N C D
3
5240



171
V H C L Q D L E D
3
5241



178
E D M S V M V L R
3
5242



181
S V M V L R T Q G
3
5243



204
S S Y D A K R A Q
3
5244



235
V S Y E V P R L H
3
5245



236
S Y E V P R L H G
3
5246



243
H G D E E R F F V
3
5247



249
F F V E G L S F P
3
5248



252
E G L S F P D A G
3
5249



266
S F H V T L L D D
3
5250



274
D S N E D F S A S
3
5251



284
I F T D T V V F R
3
5252



287
D T V V F R V A P
3
5253



317
T C F V D A V A E
3
5254



320
V D A V A E L A R
3
5255



348
I Q D E M E L G Y
3
5256



406
S F G N L E V S P
3
5257



407
F G N L E V S P P
3
5258



408
G N L E V S P P V
3
5259



463
V D W L A V C H V
3
5260



470
H V D E F L S F V
3
5261



514
F Q G V V D D E Q
3
5262



521
B Q V K T I S I N
3
5263



59
G K H L G T P K P
3
5264



611
C C L E E K V R S
3
5265



615
E K V R S L L E P
3
5266



644
V H C G T N V C R
3
5267



14
P T S A V C V A G
2
5268



15
T S A V C V A G V
2
5269



22
G V E T L V D I Y
2
5270



23
V E T L V D I Y G
2
5271



31
G S V P E G T E M
2
5272



36
G T E M F E V Y G
2
5273



40
F E V Y G T P G V
2
5274



41
E V Y G T P G V D
2
5275



47
G V D T Y I S P N
2
5276



48
V D I Y I S P N H
2
5277



57
E R G R E R A D T
2
5278



61
E R A D T R R W R
2
5279



89
D S H V Q I S Y H
2
5280



90
S H V Q I S Y H S
2
5281



94
I S Y R S S H E P
2
5282



100
H E P L P L A Y A
2
5283



112
L T C V D I S L D
2
5284



119
L D C D L N C E G
2
5285



123
L N C E G R Q D R
2
5286



175
Q D L E D M S V M
2
5287



179
D M S V M V L R T
2
5288



197
H K L V L H T S S
2
5289



203
T S S Y D A K R A
2
5290



213
V F H I C C P F D
2
5291



216
I C G P E D V C E
2
5292



220
E D V C E A Y R H
2
5293



221
D V C E A Y R H V
2
5294



228
H V L G Q D K V S
2
5295



231
G Q D K V S Y E V
2
5296



238
E V P R L H G D E
2
5297



245
D E E R F F V E G
2
5298



247
E R F F V E G L S
2
5299



251
V E G L S F P D A
2
5300



255
S F P D A G F T G
2
5301



260
G F T G L I S F H
2
5302



261
F T G L I S F H V
2
5303



268
H V T L L D D S N
2
5304



269
V T L L D D S N E
2
5305



275
S N E D F S A S P
2
5306



279
F S A S P I F T D
2
5307



285
F T D T V V F R V
2
5308



300
P S T L P P L E V
2
5309



301
S T L P P L E V Y
2
5310



315
N N T C F V D A V
2
5311



322
A V A E L A R K A
2
5312



338
P Q A E N R N D R
2
5313



350
D E M E L G Y V Q
2
5314



352
M H L G Y V Q A P
2
5315



372
R N G E L Q D F P
2
5316



388
D F G Y V T R E P
2
5317



389
F G Y V T R E P R
2
5318



400
S V S C L D S F G
2
5319



401
V S G L D S F G N
2
5320



404
L D S F G N L E V
2
5321



412
V S P P V V A N G
2
5322



415
P V V A N G K E Y
2
5323



431
G G N L P G S S G
2
5324



44
R R V T Q V V R D
2
5325



462
F V D W L A V G H
2
5326



468
V G H V D E F L S
2
5327



472
D E F L S F V P A
2
5328



476
S F V P A P D C K
2
5329



486
F R M L L A S P G
2
5330



497
F K L F Q E K Q K
2
5331



502
E K Q K C G H G R
2
5332



509
G R A L L F Q G V
2
5333



517
V V D D E Q V K T
2
5334



523
V K T I S I N Q V
2
5335



526
I S I N Q V L S N
2
5336



528
I N Q V L S N K D
2
5337



53
N K D L I N Y N K
2
5338



543
F V Q S C I D W N
2
5339



556
K R E L G L A E C
2
5340



563
E C D I I D I P Q
2
5341



588
V N M L V L G K H
2
5342



603
F C P I I N C C C
2
5343



607
I N G O C O L E E
2
5344



610
C C C L E E K V R
2
5345



617
V R S L L E P L G
2
5346



63
F T P Y H M L H G
2
5347



638
H M L H G E V H C
2
5348



640
L H G E V H C G T
2
5349



641
H G E V H C G T N
2
5350



643
E V H C G T N V C
2
5351



645
H C G T N V C R K
2
5352



7
V R V S L E H P T
1
5353



8
R V S L E H P T S
1
5354



29
I Y G S V P E G T
1
5355



30
Y G S V P E G T E
1
5356



34
P E G T E M F E V
1
5357



37
T E H F E V Y G T
1
5358



39
M F E V Y G T P G
1
5359



43
Y G T P G V D I Y
1
5360



52
I S P N M E R G R
1
5361



59
G R E R A D T R R
1
5362



70
F D A T L E I I V
1
5363



7
L E I I V V N N S
1
5364



77
I V V M N S P S N
1
5365



78
V V M N S P S N D
1
5366



80
N N S P S N D L N
1
5367



83
P S N D L N D S H
1
5368



87
L N D S H V Q I S
1
5369



91
H V Q I S Y H S S
1
5370



92
V Q I S Y H S S H
1
5371



106
A Y A V L Y L T C
1
5372



108
A V L Y L T C V D
1
5373



110
L Y L T C V D I S
1
5374



117
I S L D C D L N C
1
5375



120
D C D L N C F O R
1
5376



128
R Q D R N F V D K
1
5377



130
D R N F V D K R Q
1
5378



131
R N F V D K R Q W
1
5379



137
R Q W V W G P S G
1
5380



140
V W G P S G Y G G
1
5381



145
G Y G G I L L V N
1
5382



146
Y G G I L L V N C
1
5383



157
D D P S C D V Q D
1
5384



161
C D V Q D N C D Q
1
5385



163
V Q D N C D Q H V
1
5386



165
D N C D Q H V H C
1
5387



168
D Q H V H C L Q D
1
5388



170
H V H C L Q D L E
1
5389



172
H C L Q D L E D M
1
5390



174
L Q D L E D M S V
1
5391



195
D D H K L V L H T
1
5392



201
L H T S S Y D A K
1
5393



202
H T S S Y D A K R
1
5394



211
A Q V F H I C G P
1
5395



214
F H I C G P E D V
1
5396



219
P E D V C E A Y R
1
5397



226
Y R H V L G Q D K
1
5398



227
R H V L G Q D K V
1
5399



233
D K V S Y E V P R
1
5400



240
P R L H G D E E R
1
5401



250
F V E C L S F P D
1
5402



265
I S F H V T L L D
1
5403



267
F H V T L L D D S
1
5404



286
T D T V V F R V A
1
5405



291
F F V A P W I M T
1
5406



292
R V A P W I M T P
1
5407



29
I M T P S T L P P
1
5408



306
L E V Y V C R V R
1
5409



313
V R N N T C F V D
1
5410



31
R N N T C F V D A
1
5411



319
F V D A V A E L A
1
5412



334
L T I C P Q A E N
1
5413



336
I C P Q A E N R N
1
5414



343
R N D R W I Q D E
1
5415



354
L G Y V Q A P H K
1
5416



355
G Y V Q A P H K T
1
5417



357
V Q A P H K T L P
1
5418



362
K T L P V V F D S
1
5419



366
V V F D S P R N G
1
5420



367
V F D S P R N C E
1
5421



373
N G E L Q D F P Y
1
5422



374
G E L Q D F P Y K
1
5423



382
K R I L G P D F G
1
5424



387
P D F G Y V T R E
1
5425



398
D R S V S G L D S
1
5426



410
L E V S P P V V A
1
5427



416
V V A N G K E Y P
1
5428



430
I G G N L P G S S
1
5429



432
G N L P G S S G R
1
5430



446
V R D F L H A Q K
1
5431



447
R D F L H A Q K V
1
5432



448
D F L H A Q K V Q
1
5433



455
V Q P P V E L F V
1
5434



459
V F L F V D W L A
1
5435



471
V D E F L S F V P
1
5436



475
L S F V P A P D G
1
5437



477
F V P A P D G K G
1
5438



484
K G F R M L L A S
1
5439



487
R M L L A S P G A
1
5440



495
A C F K L F Q E K
1
5441



500
F Q E K Q K C G H
1
5442



507
G H G R A L L F Q
1
5443



513
L F Q C V V D D E
1
5444



515
Q G V V D D E Q V
1
5445



516
C V V D D F Q V K
1
5446



539
N Y N K F V Q S C
1
5447



541
N K F V Q S C I D
1
5448



544
V Q S C I D W N R
1
5449



549
D W N R E V L K R
1
5450



562
A F C D I I D I P
1
5451



567
I D I P Q L F K T
1
5452



582
A F F P D L V N M
1
5453



587
L V N M L V L G K
1
5454



602
P F G P I I N G C
1
5455



608
N G C C C L F E K
1
5456



609
G C C C L E E K V
1
5457



621
L E P L G L H C T
1
5458



627
H C T F I D D F T
1
5459



631
I D D F T P Y H M
1
5460



636
P Y H M L H G E V
1
5461



637
Y H M L H G E V H
1
5462



642
G E V H C G T N V
1
5463



652
R K P F S F K W W
1
5464



655
F S F K W W N M V
1
5465











184P1E2 v.2: HLA Peptide


Scoring Results B08 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







3
M T P S T L A P L
10
5466



9
A P L E V Y V C R
9
5467



7
T L A P L E V Y V
7
5468



4
T P S T L A P L E
6
5469



8
L A P L E V Y V C
6
5470



1
W I M T P S T L A
4
5471



2
I M T P S T L A P
2
5472



5
P S T L A P L E V
2
5473



6
S T L A P L E V Y
2
5474











184P1E2 v.3: HLA Peptide


Scoring Results B08 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







9
V P D G K G F R M
17
5475



7
V P V P D G K G F
14
5476



3
F L S F V P V P D
8
5477



2
E F L S F V P V P
3
5478



1
D E F L S F V P V
2
5479



4
L S F V P V P D G
2
5480



5
S F V P V P D C K
2
5481



6
F V P V P D G K G
2
5482

















TABLE XXX







184P1E2 v.1: HLA Peptide


Scoring Results B1510 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







360
P H K T L P V V F
21
5483



169
Q H V H C L Q D L
20
5484



242
L H G D E E R F F
19
5485



469
G H V D E F L S F
18
5486



595
K H L C I P K P F
18
5487



626
L H C T F I D D F
17
5488



453
Q K V Q P P V E L
16
5489



551
N R E V L K R E L
16
5490



99
S H F P L P L A Y
15
5491



23
K V S Y F V P R L
15
5492



377
Q D F P Y K R I L
15
5493



585
P D L V N M L V L
15
5494



612
C L E E K V R S L
15
5495



185
L R T Q G P A A L
14
5496



193
L F D D H K L V L
14
5497



222
V C E A Y R H V L
14
5498



263
G L I S F H V T L
14
5499



368
F D S P R N G E L
14
5500



421
K E Y P L G R I L
14
5501



481
P D G K G F R M L
14
5502



504
Q K C G H G R A L
14
5503



547
C I D W N R E V L
14
5504



3
L Q R I V R V S L
13
5505



12
E H P T S A V C V
13
5506



18
V C V A G V E T L
13
5507



96
Y H S S H E P L P
13
5508



142
G P S G Y G G I L
13
5509



177
L E D N S V N V L
13
5510



318
C F V D A V A E L
13
5511



356
Y V Q A P H K T L
13
5512



450
L H A Q K V Q P P
13
5513



613
L E E K V R S L L
13
5514



637
Y H M L H G E V H
13
5515



644
V H C G T N V C R
13
5516



72
A T L E I I V V H
12
5517



97
H S S H E P L P L
12
5518



102
P L P L A Y A V L
12
5519



10
P L A Y A V L Y L
12
5520



166
N C D Q H V H C L
12
5521



214
F H I C G P E D V
12
5522



246
E E R F F V E G L
12
5523



264
L I S F H V T L L
12
5524



295
P W I M T P S T L
12
5525



346
R W I Q D F N E L
12
5526



442
V T Q V V R D F L
12
5527



458
P V E L F V D W L
12
5528



482
D G K G F R M L L
12
5529



505
K C G H G R A L L
12
5530



52
K T I S I N Q V L
12
5531



553
E V L K R E L G L
12
5532



56
C D I I D I P Q L
12
5533



583
F F P D L V N M L
12
5534



6
R R W R F D A T L
11
5535



79
V M N S P S N D L
11
5536



90
S H V Q I S Y H S
11
5537



111
Y L T C V D I S L
11
5538



143
P S G Y G G I L L
11
5539



171
V H C L Q D L E D
11
5540



196
D H K L V L H T S
11
5541



201
L H T S S Y D A K
11
5542



206
Y D A K R A Q V F
11
5543



227
R H V L G Q D K V
11
5544



256
F P D A G F T G L
11
5545



298
M T P S T L P P L
11
5546



326
L A R K A G C K L
11
5547



396
P R D R S V S G L
11
5548



402
S G L D S F G N L
11
5549



417
V A N G K E Y P L
11
5550



426
G R I L I G G N L
11
5551



491
A S P G A C F K L
11
5552



507
G H G R A L L F Q
11
5553



579
K A T A F F P D L
11
5554



589
N M L V L G K H L
11
5555



605
P I I N G C C C L
11
5556



618
R S L L E P L G L
11
5557



632
D D F T P Y H M L
11
5558



640
L H G E V H C G T
11
5559



31
G S V P E G T F M
10
5560



95
S Y H S S H E P L
10
5561



115
V D I S L D C D L
10
5562



175
Q D L F D N S V M
10
5563



191
A A L F D D H K L
10
5564



267
F H V T L L D D S
10
5565



283
P I F T D T V V F
10
5566



441
R V T Q V V R D F
10
5567



454
K V Q P P V E L F
10
5568



467
A V G H V D E F L
10
5569



529
N Q V L S N K D L
10
5570



582
A F F P D L V N M
10
5571



616
K V R S L L E P L
10
5572



62
R A D T R R W R F
9
5573



241
R L H G D E E R F
9
5574



253
C L S F P D A G F
9
5575



411
E V S P P V V A N
9
5576



466
L A V G H V D E F
9
5577



478
V P A P D G K G F
9
5578



480
A P D G K G F R M
9
5579



575
T E R K K A T A F
9
5580



622
E P L G L H C T F
9
5581



631
I D D F T P Y H M
9
5582



647
G T N V C R K P F
9
5583



32
S V P E G T E M F
8
5584



51
Y I S P N M E R G
8
5585



125
C E G R Q D R N F
8
5586



186
R T Q G P A A L F
8
5587



277
E D F S A S P I F
8
5588



290
V F R V A P W I M
8
5589



381
Y K R I L G P D F
8
5590



489
L L A S P G A C F
8
5591



565
D I I D I P Q L F
8
5592



576
H R K K A T A F F
8
5593



48
V D I Y I S P N M
7
5594



172
H C L Q D L E D M
7
5595



248
R F F V E C L S F
7
5596



259
A G F T G L I S F
7
5597



271
L L D D S N E D F
7
5598



307
E V Y V C R V R N
7
5599



311
C R V R N N T C F
7
5600



344
N D R W I Q D E M
7
5601



371
P R N G H L Q D F
7
5602



399
R S V S G L D S F
7
5603



435
P G S S G R R V T
7
5604



436
G S S C R R V T Q
7
5605



506
C G H G R A L L F
7
5606



535
K D L I N Y N K F
7
5607



649
N V C R K P F S F
7
5608



654
P F S F K W W N N
7
5609



1
M S L Q R I V R V
6
5610



61
E R A D T R R W R
6
5611



149
I L L V N C D R D
6
5612



216
I C G P E D V C E
6
5613



284
I F T D T V V F R
6
5614



286
T D T V V F R V A
6
5615



393
T R E P R D R S V
6
5616



409
N L H V S P P V V
6
5617



410
L E V S P P V V A
6
5618



439
G R R V T Q V V R
6
5619



492
S P G A C F K L F
6
5620



611
C C L E E K V R S
6
5621



2
S L Q R I V R V S
5
5622



30
Y G S V P E G T E
5
5623



41
E V Y G T P G V D
5
5624



43
Y G T P G V D I Y
5
5625



55
N M E R G R E R A
5
5626



73
T L H I I V V N N
5
5627



179
D M S V M V L R T
5
5628



204
S S Y D A K R A Q
5
5629



244
G D E E R F F V E
5
5630



287
D T V V F R V A P
5
5631



292
R V A P W I M T P
5
5632



301
S T L P P L E V Y
5
5633



302
T L P P L E V Y V
5
5634



305
P L E V Y V C R V
5
5635



317
T C F V D A V A E
5
5636



336
I C P Q A E N R N
5
5637



351
E N H L C Y V Q A
5
5638



386
G P D F G Y V T R
5
5639



392
V T R H P R D R S
5
5640



420
G K E Y P L G R I
5
5641



440
R R V T Q V V R D
5
5642



452
A Q K V Q P P V E
5
5643



465
W L A V G H V D E
5
5644



512
L L F Q G V V D D
5
5645



525
T I S I N Q V L S
5
5646



599
I P K P F G P I I
5
5647



11
L E H P T S A V C
4
5648



14
P T S A V C V A G
4
5649



15
T S A V C V A G V
4
5650



17
A V C V A G V E T
4
5651



28
D I Y G S V P E G
4
5652



35
E G T E M F E V Y
4
5653



36
G T H M F E V Y G
4
5654



50
I Y I S P N M E R
4
5655



52
I S P N M H R G R
4
5656



54
P N M E R G R E R
4
5657



56
M E R G R E R A D
4
5658



59
G R E R A D T R R
4
5659



60
R E R A D T R R W
4
5660



77
I V V M N S P S N
4
5661



101
E P L P L A Y A V
4
5662



124
N C E C R Q D R N
4
5663



127
C R Q D R N F V D
4
5664



131
R N F V D K R Q W
4
5665



145
C Y G G I L L V N
4
5666



156
R D D P S C D V Q
4
5667



176
D L E D M S V M V
4
5668



178
E D M S V M V L R
4
5669



203
T S S Y D A K R A
4
5670



215
H I C G P E D V C
4
5671



218
G P E D V C E A Y
4
5672



229
V L G Q D K V S Y
4
5673



235
V S Y E V P R L H
4
5674



236
S Y E V P R L H G
4
5675



245
D E E R F F V E G
4
5676



279
F S A S P I F T D
4
5677



297
I M T P S T L P P
4
5678



299
T P S T L P P L E
4
5679



306
L E V Y V C R V R
4
5680



322
A V A S L A R K A
4
5681



328
R K A G C K L T I
4
5682



338
P Q A E N R N D R
4
5683



357
V Q A P H K T L P
4
5684



359
A P H K T L P V V
4
5685



361
H K T L P V V F D
4
5686



366
V V F D S P R N G
4
5687



384
I L G P D F G Y V
4
5688



387
P D F G Y V T R E
4
5689



390
G Y V T R E P R D
4
5690



391
Y V T R E P R D R
4
5691



395
E P R D R S V S G
4
5692



405
D S F G N L E V S
4
5693



428
I L I G G N L P G
4
5694



432
C N L P G S S G R
4
5695



434
L P G S S C R R V
4
5696



473
E F L S F V P A P
4
5697



490
L A S P G A C F K
4
5698



511
A L L F Q G V V D
4
5699



516
G V V D D E Q V K
4
5700



537
L I N Y N K F V Q
4
5701



538
I N Y N K F V Q S
4
5702



546
S C I D W N R E V
4
5703



548
I D W N R K V L K
4
5704



567
I D I P Q L F K T
4
5705



569
I P Q L F K T E R
4
5706



581
T A F F P D L V N
4
5707



594
G K H L G I P K P
4
5708



643
E V H C G T N V C
4
5709



645
H C G T N V C R K
4
5710



6
I V R V S L E H P
3
5711



8
R V S L E H P T S
3
5712



10
S L E H P T S A V
3
5713



13
H P T S A V C V A
3
5714



20
V A G V E T L V D
3
5715



21
A G V E T L V D I
3
5716



22
G V E T L V D I Y
3
5717



24
E T L V D I Y G S
3
5718



27
V D I Y G S V P E
3
5719



29
I Y G S V P E G T
3
5720



38
E M F E V Y G T P
3
5721



42
V Y G T P G V D I
3
5722



45
T P G V D I Y I S
3
5723



53
S P N M E R G R E
3
5724



71
D A T L E T I V V
3
5725



75
E I I V V M N S P
3
5726



76
I I V V N N S P S
3
5727



80
M N S P S N D L N
3
5728



86
D L N D S H V Q I
3
5729



94
I S Y H S S H E P
3
5730



98
S S H E P L P L A
3
5731



106
A Y A V L Y L T C
3
5732



117
I S L D C D L N C
3
5733



123
L N C S G R Q D R
3
5734



126
E G R Q D R N F V
3
5735



128
R Q D R N F V D K
3
5736



133
F V D K R Q W V W
3
5737



144
S G Y G G I L L V
3
5738



146
Y G G I L L V N C
3
5739



165
D N C D Q H V H C
3
5740



180
M S V N V L R T Q
3
5741



187
T Q C P A A L F D
3
5742



217
C G P E D V C E A
3
5743



220
E D V C E A Y R H
3
5744



221
D V C E A Y R H V
3
5745



223
C E A Y R H V L G
3
5746



228
H V L G Q D K V S
3
5747



231
G Q D K V S Y E V
3
5748



232
Q D K V S Y E V P
3
5749



237
Y E V P R L H G D
3
5750



252
E G L S F P D A G
3
5751



260
G F T G L I S F H
3
5752



262
T G B I S F H V T
3
5753



270
T L L D D S N E D
3
5754



280
S A S P I F T D T
3
5755



285
F T D T V V F R V
3
5756



300
P S T L P P L E V
3
5757



303
L P P B E V Y V C
3
5758



304
P P L E V Y V C R
3
5759



308
V Y V C R V R N N
3
5760



320
V D A V A E L A R
3
5761



323
V A E L A R K A G
3
5762



335
T I C P Q A E N R
3
5763



339
Q A E N R N D R W
3
5764



340
A E N R N D R W I
3
5765



348
I Q D E M E L C Y
3
5766



350
D E M E L G Y V Q
3
5767



352
M E L C Y V Q A P
3
5768



355
G Y V Q A P H K T
3
5769



362
K T L P V V F D S
3
5770



365
P V V F D S P R N
3
5771



374
C E L Q D F P Y K
3
5772



375
E L Q D F P Y K R
3
5773



376
L Q D F P Y K R I
3
5774



385
L G P D F G Y V T
3
5775



388
D F G Y V T R E P
3
5776



394
R E P R D R S V S
3
5777



404
L D S F G N L E V
3
5778



408
C N L E V S P P V
3
5779



412
V S P P V V A N G
3
5780



416
V V A N C K E Y P
3
5781



422
E Y P L G R I L I
3
5782



423
Y P L C R I L I G
3
5783



429
L I C C N L P G S
3
5784



430
I G C N L P G S S
3
5785



433
N L P C S S G R R
3
5786



449
F L H A Q K V Q P
3
5787



456
Q P P V E L F V D
3
5788



457
P P V E L F V D W
3
5789



460
E L F V D W L A V
3
5790



461
L F V D W L A V G
3
5791



471
V D E F L S F V P
3
5792



475
L S F V P A P D G
3
5793



476
S F V P A P D C K
3
5794



479
P A P D C K C F R
3
5795



500
F Q E K Q K C C H
3
5796



503
K Q K C G H C R A
3
5797



517
V V D D E Q V K T
3
5798



518
V D D E Q V K T I
3
5799



519
D D E Q V K T I S
3
5800



521
E Q V K T I S I N
3
5801



526
I S I N Q V L S N
3
5802



528
I N Q V L S N K D
3
5803



550
W N R E V L K R E
3
5804



556
K R E L G L A E C
3
5805



560
G L A E C D I I D
3
5806



566
I I D I P Q L F K
3
5807



571
Q L F K T E R K K
3
5808



573
F K T E R K K A T
3
5809



574
K T E R K K A T A
3
5810



596
H L C I P K P F G
3
5811



597
L G I P K P F G P
3
5812



598
C I P K P F G P I
3
5813



601
K P F G P I I N G
3
5814



602
P F G P I I N G C
3
5815



606
I I N G C C C L E
3
5816



607
I N G C C C L E E
3
5817



620
L L E P L G L H C
3
5818



638
H M L H G E V H C
3
5819



641
H G E V H C C T N
3
5820



648
T N V C R K P F S
3
5821



4
Q R I V R V S L E
2
5822



9
V S L E H P T S A
2
5823



16
S A V C V A C V E
2
5824



19
C V A G V E T L V
2
5825



25
T L V D I Y G S V
2
5826



26
L V D I Y G S V P
2
5827



33
V P E G T E M F E
2
5828



34
P E G T E M F E V
2
5829



37
T E M F E V Y G T
2
5830



39
M F E V Y C T P G
2
5831



40
F E V Y C T P G V
2
5832



44
C T P C V D I Y I
2
5833



47
G V D I Y I S P N
2
5834



57
E R C R E R A D T
2
5835



58
R G R E R A D T R
2
5836



63
A D T R R W R F D
2
5837



65
T R R W R F D A T
2
5838



68
W R F D A T L E I
2
5839



85
N D L N D S H V Q
2
5840



89
D S H V Q I S Y H
2
5841



103
L P L A Y A V L Y
2
5842



108
A V L Y L T C V D
2
5843



109
V L Y L T C V D I
2
5844



112
L T C V D I S L D
2
5845



113
T C V D I S L D C
2
5846



119
L D C D L N C E G
2
5847



121
C D L N C E C K Q
2
5848



122
D L N C F G R Q D
2
5849



130
D R N F V D K R Q
2
5850



132
N F V D K R Q W V
2
5851



134
V D K R Q W V W G
2
5852



135
D K R Q W V W G P
2
5853



137
R Q W V W G P S G
2
5854



138
Q W V W G P S G Y
2
5855



139
W V W G P S G Y G
2
5856



140
V W G P S G Y G G
2
5857



147
G G T L L V N C D
2
5858



150
L L V N C D R D D
2
5859



154
C D R D D P S C D
2
5860



155
D R D D P S C D V
2
5861



157
D D P S C D V Q D
2
5862



158
D P S C D V Q D N
2
5863



162
D V Q D N C D Q H
2
5864



164
Q D N C D Q H V H
2
5865



181
S V M V L R T Q G
2
5866



184
V L R T Q G P A A
2
5867



189
G P A A L F D D H
2
5868



194
F D D H K L V L H
2
5869



195
D D H K L V L H T
2
5870



207
D A K R A Q V F H
2
5871



209
K R A Q V F H I C
2
5872



213
V F H I C G P E D
2
5873



224
E A Y R H V L G Q
2
5874



226
Y R H V L G Q D K
2
5875



230
L G Q D K V S Y E
2
5876



233
D K V S Y E V P R
2
5877



240
P R L H G D E E R
2
5878



243
H G D E E R F F V
2
5879



249
F F V E G L S F P
2
5880



250
F V E G L S F P D
2
5881



254
L S F P D A G F T
2
5882



255
S F P D A G F T G
2
5883



257
P D A G F T G L I
2
5884



265
I S F H V T L L D
2
5885



274
D S N E D F S A S
2
5886



275
S N E D F S A S P
2
5887



278
D F S A S P I F T
2
5888



282
S P I F T D T V V
2
5889



288
T V V F R V A P W
2
5890



309
Y V C R V R N N T
2
5891



310
V C R V R N N T C
2
5892



313
V R N N T C F V D
2
5893



314
R N N T C F V D A
2
5894



315
N N T C F V D A V
2
5895



316
N T C F V D A V A
2
5896



321
D A V A E L A R K
2
5897



325
E L A R K A G C K
2
5898



331
G C K L T I C P Q
2
5899



333
K L T I C P Q A E
2
5900



334
L T I C P Q A E N
2
5901



337
C P Q A E N R N D
2
5902



341
E N R N D R W I Q
2
5903



347
W I Q D E M F L G
2
5904



354
L G Y V Q A P H K
2
5905



363
T L P V V F D S P
2
5906



367
V F D S P R N G E
2
5907



379
F P Y K R I L G P
2
5908



382
K R I L G P D F G
2
5909



383
R I L G P D F G Y
2
5910



389
F G Y V T R E P R
2
5911



398
D R S V S G L D S
2
5912



400
S V S G L D S F G
2
5913



406
S F G N L E V S P
2
5914



415
P V V A N G K E Y
2
5915



418
A N G K E Y P L G
2
5916



419
N G K E Y P L G R
2
5917



431
G G N L P G S S G
2
5918



438
S G R R V T Q V V
2
5919



443
T Q V V R D F L H
2
5920



445
V V R D F L H A Q
2
5921



448
D F L H A Q K V Q
2
5922



451
H A Q K V Q P P V
2
5923



455
V Q P P V E L F V
2
5924



462
F V D W L A V G H
2
5925



464
D W L A V G H V D
2
5926



472
D E F L S F V P A
2
5927



474
F L S F V P A P D
2
5928



483
G K G F R M L L A
2
5929



488
M L L A S P G A C
2
5930



493
P G A C F K L F Q
2
5931



495
A C F K L F Q E K
2
5932



498
K L F Q E K Q K C
2
5933



502
E K Q K C G H G R
2
5934



509
G R A L L F Q G V
2
5935



510
R A L L F Q G V V
2
5936



513
L F Q G V V D D E
2
5937



514
F Q G V V D D E Q
2
5938



530
Q V L S N K D L I
2
5939



533
S N K D L I N Y N
2
5940



539
N Y N K F V Q S C
2
5941



545
Q S C I D W N R E
2
5942



555
L K R E L C L A E
2
5943



561
L A H C D I I D I
2
5944



562
A E C D I I D I P
2
5945



568
D I P Q L F K T E
2
5946



570
P Q L F K T E R K
2
5947



580
A T A F F P D L V
2
5948



586
D L V N M L V L G
2
5949



590
M L V L G K H L G
2
5950



600
P K P F G P I I N
2
5951



60
G P I I N G C C C
2
5952



61
C C C L H E K V R
2
5953



61
E E K V R S L L E
2
5954



615
E K V R S L L E P
2
5955



617
V R S L L E P L G
2
5956



619
S L L E P L G L H
2
5957



629
T F I D D F T P Y
2
5958



630
F I D D F T P Y H
2
5959



633
D F T P Y H M L H
2
5960



639
M L H G E V H C G
2
5961



646
C G T N V C R K P
2
5962



650
V C R K P F S F K
2
5963



651
C R K P F S F K W
2
5964



656
S F K W W N M V P
2
5965



5
R I V R V S L E H
1
5966



46
P G V D I Y I S P
1
5967



49
D I Y I S P N M E
1
5968



64
D T R R W R F D A
1
5969



70
F D A T L E I I V
1
5970



74
L E I I V V M N S
1
5971



78
V V M N S P S N D
1
5972



81
N S P S N D L N D
1
5973



82
S P S N D L N D S
1
5974



83
P S N D L N D S H
1
5975



84
S N D L N D S H V
1
5976



87
L N D S H V Q I S
1
5977



88
N D S H V Q I S Y
1
5978



93
Q I S Y H S S H E
1
5979



100
H E P L P L A Y A
1
5980



107
Y A V L Y L T C V
1
5981



110
L Y L T C V D I S
1
5982



116
D I S L D C D L N
1
5983



120
D C D L N C H C R
1
5984



129
Q D R N F V D K R
1
5985



141
W C P S G Y G G I
1
5986



148
G I L L V N C D R
1
5987



151
L V N C D R D D P
1
5988



152
V N C D R D D P S
1
5989



153
N C D R D D P S C
1
5990



159
P S C D V Q D N C
1
5991



160
S C D V Q D N C D
1
5992



161
C D V Q D N C D Q
1
5993



168
D Q H V H C L Q D
1
5994



173
C L Q D L E D M S
1
5995



190
P A A L F D D H K
1
5996



192
A L F D D H K L V
1
5997



197
H K L V L H T S S
1
5998



202
H T S S Y D A K R
1
5999



205
S Y D A K R A Q V
1
6000



208
A K R A Q V F H I
1
6001



210
R A Q V F H I C C
1
6002



211
A Q V F H I C G P
1
6003



219
P E D V C H A Y R
1
6004



238
H V P R L H G D E
1
6005



239
V P R L H G D E E
1
6006



247
E R F F V E G L S
1
6007



251
V E G L S F P D A
1
6008



266
S F H V T L L D D
1
6009



272
L D D S N E D F S
1
6010



273
D D S N E D F S A
1
6011



281
A S P I F T D T V
1
6012



289
V V F R V A P W I
1
6013



291
F R V A P W I M T
1
6014



293
V A P W I M T P S
1
6015



294
A P W I M T P S T
1
6016



296
W I M T P S T L P
1
6017



312
R V R N N T C F V
1
6018



324
A E L A R K A G C
1
6019



327
A R K A G C K L T
1
6020



330
A G C K L T I C P
1
6021



332
C K L T I C P Q A
1
6022



342
N R N D R W I Q D
1
6023



343
R N D R W I Q D E
1
6024



349
Q D E M E L G Y V
1
6025



353
E L G Y V Q A P H
1
6026



364
L P V V F D S P R
1
6027



369
D S P R N G E L Q
1
6028



370
S P R N G E L Q D
1
6029



372
R N C E L Q D F P
1
6030



373
N C H L Q D F P Y
1
6031



378
D F P Y K R I L C
1
6032



403
C L D S F C N L E
1
6033



407
F G N L E V S P P
1
6034



413
S P P V V A N G K
1
6035



414
P P V V A N G K E
1
6036



425
L G R I L I G G N
1
6037



427
R I L I G G N L P
1
6038



437
S S G R R V T Q V
1
6039



463
V D W L A V G H V
1
6040



468
V G H V D E F L S
1
6041



470
H V D E F L S F V
1
6042



477
F V P A P D G K G
1
6043



484
K G F R M L L A S
1
6044



485
G F R M L L A S P
1
6045



487
R M L L A S P G A
1
6046



494
G A C F K L F Q E
1
6047



499
L F Q E K Q K C G
1
6048



501
Q E K Q K C G H G
1
6049



515
Q G V V D D E Q V
1
6050



520
D E Q V K T I S I
1
6051



523
V K T I S I N Q V
1
6052



527
S I N Q V L S N K
1
6053



531
V L S N K D L I N
1
6054



532
L S N K D L I N Y
1
6055



540
Y N K F V Q S C I
1
6056



541
N K F V Q S C I D
1
6057



542
K F V Q S C I D W
1
6058



543
F V Q S C I D W N
1
6059



544
V Q S C I D W N R
1
6060



549
D W N R E V L K R
1
6061



552
R E V L K R E L G
1
6062



557
R E L G L A E C D
1
6063



558
E L G L A E C D I
1
6064



563
E C D I I D I P Q
1
6065



572
L F K T E R K K A
1
6066



578
K K A T A F F P D
1
6067



587
L V N M L V L G K
1
6068



591
L V L G K H L G I
1
6069



592
V L G K H L G I P
1
6070



608
N G C C C L E E K
1
6071



609
G C C C L E E K V
1
6072



621
L E P L G L H C T
1
6073



625
G L H C T F I D D
1
6074



628
C T F I D D F T P
1
6075



634
F T P Y H M L H G
1
6076



635
T P Y H M L H G E
1
6077



636
P Y H M L H G E V
1
6078



642
G E V H C G T N V
1
6079



653
K P F S F K W W N
1
6080



655
F S F K W W N M V
1
6081











184P1E2 v.2: HLA Peptide


Scoring Results B1510 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







3
M T P S T L A P L
10
6082



7
T L A P L E V Y V
6
6083



2
I M T P S T L A P
5
6084



4
T P S T L A P L E
3
6085



6
S T L A P L E V Y
3
6086



8
L A P L E V Y V C
3
6087



9
A P L E V Y V C R
3
6088



5
P S T L A P L E V
2
6089



1
W I M T P S T L A
1
6090











184P1E2 v.3: HLA Peptide


Scoring Results B1510 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







9
V P D G K G F R M
9
6091



7
V P V P D G K G F
8
6092



2
E F L S F V P V P
5
6093



3
F L S F V P V P D
4
6094



4
L S F V P V P D G
4
6095



5
S F V P V P D G K
3
6096



8
P V P D G K G F R
3
6097



1
D E F L S F V P V
2
6098



6
F V P V P D G K G
2
6099

















TABLE XXXI







184P1E2 v.1: HLA Peptide


Scoring Results B2705 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







426
G R I L I G G N L
29
6100



59
G R E R A D T R R
28
6101



66
R R W R F D A T L
27
6102



439
G R R V T Q V V R
27
6103



240
P R L H G D E E R
26
6104



185
L R T Q G P A A L
25
6105



311
C R V R N N T C F
24
6106



371
P R N G E L Q D F
24
6107



396
P R D R S V S G L
23
6108



446
V R D F L H A Q K
23
6109



551
N R E V L K R E L
23
6110



576
E R K K A T A F F
23
6111



68
W R F D A T L E I
21
6112



226
Y R H V L G Q D K
21
6113



61
E R A D T R R W R
20
6114



248
R F F V E G L S F
20
6115



346
R W I Q D E M E L
20
6116



432
G N L P G S S G R
20
6117



186
R T Q G P A A L F
19
6118



259
A G F T C L I S F
19
6119



399
R S V S G L D S F
19
6120



440
R R V T Q V V R D
19
6121



618
R S L L E P L C L
19
6122



72
A T L F I I V V M
18
6123



148
G I L L V N C D R
18
6124



241
R L H G D E E R F
18
6125



260
G F T C L I S F H
18
6126



374
G E L Q D F P Y K
18
6127



383
R I L G P D F G Y
18
6128



441
R V T Q V V R D F
18
6129



495
A C F K L F Q E K
18
6130



556
K R E L G L A E C
18
6131



5
R I V R V S L E H
17
6132



31
G S V P E G T E M
17
6133



50
I Y I S P N M E R
17
6134



58
R G R E R A D T R
17
6135



62
R A D T R R W R F
17
6136



234
K V S Y F V P R L
17
6137



263
C L I S F H V T L
17
6138



382
K R I L C P D F G
17
6139



386
G P D F G Y V T R
17
6140



469
G H V D F F L S F
17
6141



524
K T I S I N Q V L
17
6142



527
S I N Q V L S N K
17
6143



535
K D L I N Y N K F
17
6144



564
C D I I D I P Q L
17
6145



571
Q L F K T E R K K
17
6146



582
A F F P D L V N M
17
6147



595
K H L C I P K P F
17
6148



191
A A L F D D H K L
16
6149



253
G L S F P D A G F
16
6150



321
D A V A E L A R K
16
6151



421
K E Y P L G R I L
16
6152



433
N L P G S S G R R
16
6153



453
Q K V Q P P V E L
16
6154



534
N K D L I N Y N K
16
6155



569
I P Q L F K T E R
16
6156



575
T E R K K A T A F
16
6157



622
E P L G L H C T F
16
6158



18
V C V A G V E T L
15
6159



48
V D I Y I S P N M
15
6160



127
G R Q D R N F V D
15
6161



128
R Q D R N F V D K
15
6162



142
G P S G Y G C I L
15
6163



209
K R A Q V F H I C
15
6164



277
E D F S A S P I F
15
6165



283
P I F T D T V V F
15
6166



304
P P L E V Y V C R
15
6167



328
R K A G C K L T I
15
6168



377
Q D F P Y K R I L
15
6169



402
S G L D S F G N L
15
6170



454
K V Q P P V E L F
15
6171



466
L A V G H V D E F
15
6172



491
A S P G A C F K L
15
6173



505
K C G H G R A L L
15
6174



509
G R A L L F Q G V
15
6175



516
G V V D D E Q V K
15
6176



544
V Q S C I D W N R
15
6177



549
D W N R E V L K R
15
6178



553
E V L K R E L G L
15
6179



566
I I D I P Q L F K
15
6180



570
P Q L F K T E R K
15
6181



4
Q R I V R V S L E
14
6182



44
G T P G V D I Y I
14
6183



111
Y L T C V D I S L
14
6184



115
V D I S L D C D L
14
6185



130
D R N F V D K R Q
14
6186



177
L E D M S V M V L
14
6187



198
K L V L H T S S Y
14
6188



202
H T S S Y D A K R
14
6189



206
Y D A K R A Q V F
14
6190



220
E D V C E A Y R H
14
6191



247
E R F F V E G L S
14
6192



256
F P D A G F T G L
14
6193



295
P W I M T P S T L
14
6194



301
S T L P P L F V Y
14
6195



318
C F V D A V A E L
14
6196



326
L A R K A G C K L
14
6197



335
T I C P Q A E N R
14
6198



354
L G Y V Q A P H K
14
6199



417
V A N G K F Y P L
14
6200



480
A P D C K G F R M
14
6201



481
P D C K G F R N L
14
6202



486
F R M L L A S P G
14
6203



490
L A S P C A C F K
14
6204



506
C G H C R A L L F
14
6205



529
N Q V L S N K D L
14
6206



532
L S N K D L I N Y
14
6207



585
P D L V N M L V L
14
6208



601
K P F G P I I N G
14
6209



605
P I I N G C C C L
14
6210



612
C L E E K V R S L
14
6211



613
L E E K V R S L L
14
6212



616
K V R S L L S P L
14
6213



632
D D F T P Y H M L
14
6214



644
V H C G T N V C R
14
6215



650
V C R K P F S F K
14
6216



651
C R K P F S F K W
14
6217



3
L Q R I V R V S L
13
6218



22
G V E T L V D I Y
13
6219



32
S V P E C T E M F
13
6220



79
V M N S P S N D L
13
6221



88
N D S H V Q I S Y
13
6222



102
P L P L A Y A V L
13
6223



123
L N C E G R Q D R
13
6224



125
C E G R Q D R N F
13
6225



129
Q D R N F V D K R
13
6226



136
K R Q W V W G P S
13
6227



143
P S G Y C C I L L
13
6228



166
N C D Q H V H C L
13
6229



172
H C L Q D L E D M
13
6230



175
Q D L E D M S V H
13
6231



193
L F D D H K L V L
13
6232



229
V L G Q D K V S Y
13
6233



264
L I S F H V T L L
13
6234



284
I F T D T V V F R
13
6235



291
F R V A P W I M T
13
6236



306
L E V Y V C R V R
13
6237



327
A R K A G C K L T
13
6238



360
P H K T L P V V F
13
6239



364
L P V V F D S P R
13
6240



368
F D S P R N C K L
13
6241



375
E L Q D F P Y K R
13
6242



393
T R E P R D R S V
13
6243



413
S P P V V A N C K
13
6244



420
G K E Y P L G R I
13
6245



467
A V C H V D E F L
13
6246



479
P A P D G K G F R
13
6247



482
D G K G F R M L L
13
6248



497
F K L F Q E K Q K
13
6249



565
D I I D I P Q L F
13
6250



579
K A T A F F P D L
13
6251



583
F F P D L V N M L
13
6252



587
L V N M L V L C K
13
6253



588
V N M L V L G K H
13
6254



589
N H L V L C K H L
13
6255



610
C C C L S S K V R
13
6256



617
V R S L L E P L G
13
6257



619
S L L E P L G L H
13
6258



626
L H C T F I D D F
13
6259



645
H C C T N V C R K
13
6260



647
G T N V C R K P F
13
6261



7
V R V S L E H P T
12
6262



43
Y G T P G V D I Y
12
6263



5
P N M E R G R E R
12
6264



57
E R C R E R A D T
12
6265



89
D S H V Q I S Y H
12
6266



92
V Q I S Y H S S H
12
6267



97
H S S H E P L P L
12
6268



103
L P L A Y A V L Y
12
6269



104
P L A Y A V L Y L
12
6270



138
Q W V W G P S G Y
12
6271



155
D R D D P S C D V
12
6272



169
Q H V H C L Q D L
12
6273



178
E D M S V N V L R
12
6274



189
G P A A L F D D H
12
6275



190
P A A L F D D H K
12
6276



218
G P E D V C E A Y
12
6277



227
R H V L G Q D K V
12
6278



233
D K V S Y E V P R
12
6279



235
V S Y F V P R L H
12
6280



271
L L D D S N E D F
12
6281



289
V V F R V A P W I
12
6282



298
M T P S T L P P L
12
6283



325
E L A R K A C C K
12
6284



338
P Q A H N R N D R
12
6285



342
N R N D R W I Q D
12
6286



344
N D R W I Q D F M
12
6287



353
E L C Y V Q A P H
12
6288



356
Y V Q A P H K T L
12
6289



381
Y K R I L G P D F
12
6290



389
F G Y V T R S P R
12
6291



398
D R S V S G L D S
12
6292



419
N G K E Y P L G R
12
6293



458
P V E L F V D W L
12
6294



476
S F V P A P D G K
12
6295



489
L L A S P G A C F
12
6296



502
E K Q K C G H G R
12
6297



504
Q K C G H G R A L
12
6298



548
I D W N R E V L K
12
6299



591
L V L G K H L G I
12
6300



593
L G K H L G I P K
12
6301



594
G K H L G I P K P
12
6302



598
C I P K P F G P I
12
6303



608
N G C C C L E E K
12
6304



629
T F I D D F T P Y
12
6305



649
N V C R K P F S F
12
6306



654
P F S F K W W N M
12
6307



21
A G V E T L V D I
11
6308



52
I S P N M E R G R
11
6309



69
R F D A T L E I I
11
6310



83
P S N D L N D S H
11
6311



95
S Y H S S H E P L
11
6312



99
S H E P L P L A Y
11
6313



120
D C D L N C E C R
11
6314



164
Q D N C D Q H V H
11
6315



194
F D D H K L V L H
11
6316



201
L H T S S Y D A K
11
6317



207
D A K R A Q V F H
11
6318



219
P E D V C E A Y R
11
6319



222
V C E A Y R H V L
11
6320



242
L H G D E E R F F
11
6321



246
E E R F F V E G L
11
6322



313
V R N N T C F V D
11
6323



320
V D A V A E L A R
11
6324



345
D R W I Q D E M E
11
6325



348
I Q D E M E L G Y
11
6326



391
Y V T R E P R D R
11
6327



442
V T Q V V R D F L
11
6328



447
R D F L H A Q K V
11
6329



462
F V D W L A V G H
11
6330



478
V P A P D G K G F
11
6331



498
K L F Q E K Q K C
11
6332



500
F Q E K Q K C G H
11
6333



547
C I D W N R E V L
11
6334



1
M S L Q R I V R V
10
6335



35
E G T E M F E V Y
10
6336



65
T R R W R F D A T
10
6337



109
V L Y L T C V D I
10
6338



144
S C Y G G I L L V
10
6339



292
R V A P W I M T P
10
6340



373
N C E L Q C F P Y
10
6341



387
P D F C Y V T R E
10
6342



415
P V V A N G K E Y
10
6343



422
E Y P L G R I L I
10
6344



443
T Q V V R D F L H
10
6345



492
S P G A C F K L F
10
6346



510
R A L L F Q C V V
10
6347



540
Y N K F V Q S C I
10
6348



557
R E L G L A E C D
10
6349



561
L A E C D I I D I
10
6350



631
I D D F T P Y H M
10
6351



633
D F T P Y H M L H
10
6352



637
Y H M L H C E V H
10
6353



86
D L N D S H V Q I
9
6354



131
R N F V D K R Q W
9
6355



145
G Y G G I L L V N
9
6356



147
G G I L L V N C D
9
6357



162
D V Q D N C D Q H
9
6358



231
G Q D K V S Y E V
9
6359



276
N E D F S A S P I
9
6360



290
V F R V A P W I M
9
6361



376
L Q D F P Y K R I
9
6362



408
C N L E V S P P V
9
6363



427
R I L I G G N L P
9
6364



428
T L I G G N L P G
9
6365



487
R M L L A S P C A
9
6366



512
L L F Q G V V D D
9
6367



518
V D D E Q V K T I
9
6368



520
D E Q V K T I S I
9
6369



530
Q V L S N K D L I
9
6370



559
L G L A E C D I I
9
6371



599
I P K P F C P I I
9
6372



623
P L G L H C T F I
9
6373



630
F I D D F T P Y H
9
6374



8
R V S L E H P T S
8
6375



28
D I Y C S V P E G
8
6376



42
V Y G T P G V D I
8
6377



141
W G P S G Y C C I
8
6378



208
A K R A Q V F H I
8
6379



257
P D A C F T G L I
8
6380



340
A E N R N D R W I
8
6381



355
G Y V Q A P H K T
8
6382



372
R N C E L Q D F P
8
6383



379
F P Y K R I L G P
8
6384



423
Y P L G K I L I C
8
6385



611
C C L E E K V R S
8
6386



642
C E V H C G T N V
8
6387



47
G V D I Y I S P N
7
6388



67
R W R F D A T L E
7
6389



71
D A T L E I I V V
7
6390



74
L E I I V V M N S
7
6391



117
I S L D C D L N C
7
6392



156
R D D P S C D V Q
7
6393



269
V T L L D D S N E
7
6394



331
G C K L T I C P Q
7
6395



334
L T I C P Q A E N
7
6396



343
R N D R W I Q D E
7
6397



362
K T L P V V F D S
7
6398



390
G Y V T R E P R D
7
6399



405
D S F G N L E V S
7
6400



431
G G N L P G S S G
7
6401



436
G S S G R R V T Q
7
6402



484
K G F R M L L A S
7
6403



485
G F R M L L A S P
7
6404



521
E Q V K T I S I N
7
6405



542
K F V Q S C I D W
7
6406



552
R E V L K R E L G
7
6407



558
E L G L A E C D I
7
6408



567
I D I P Q L F K T
7
6409



581
T A F F P D L V N
7
6410



653
K P F S F K W W N
7
6411



9
V S L E H P T S A
6
6412



24
E T L V D I Y G S
6
6413



27
V D I Y G S V P E
6
6414



38
E M F E V Y G T P
6
6415



60
R E R A D T R R W
6
6416



90
S H V Q I S Y H S
6
6417



94
I S Y H S S H E P
6
6418



101
E P L P L A Y A V
6
6419



124
N C E G R Q D R N
6
6420



133
F V D K R Q W V W
6
6421



137
R Q W V W G P S G
6
6422



197
H K L V L H T S S
6
6423



228
H V L G Q D K V S
6
6424



249
F F V E G L S F P
6
6425



265
I S F H V T L L D
6
6426



282
S P I F T D T V V
6
6427



322
A V A E L A R K A
6
6428



329
K A G C K L T I C
6
6429



336
I C P Q A E N R N
6
6430



359
A P H K T L P V V
6
6431



366
V V F D S P R N G
6
6432



394
R E P R D R S V S
6
6433



397
R D R S V S G L D
6
6434



412
V S P P V V A N G
6
6435



451
H A Q K V Q P P V
6
6436



472
D E F L S F V P A
6
6437



475
L S F V P A P D G
6
6438



494
G A C F K L F Q E
6
6439



503
K Q K C G H G R A
6
6440



517
V V D D E Q V K T
6
6441



519
D D E Q V K T I S
6
6442



523
V K T I S I N Q V
6
6443



526
I S I N Q V L S N
6
6444



528
I N Q V L S N K D
6
6445



538
I N Y N K F V Q S
6
6446



560
G L A E C D I I D
6
6447



57
K T E R K K A T A
6
6448



602
P F G P I I N G C
6
6449



609
G C C C L E E K V
6
6450



655
F S F K W W N M V
6
6451



19
C V A G V E T L V
5
6452



41
E V Y G T P G V D
5
6453



75
E I I V V M N S P
5
6454



77
I V V M N S P S N
5
6455



105
L A Y A V L Y L T
5
6456



107
Y A V L Y L T C V
5
6457



108
A V L Y L T C V D
5
6458



146
Y G G I L L V N C
5
6459



149
I L L V N C D R D
5
6460



159
P S C D V Q D N C
5
6461



174
L Q D L E D M S V
5
6462



195
D D H K L V L H T
5
6463



210
R A Q V F H I C G
5
6464



211
A Q V F H I C G P
5
6465



216
I C G P E D V C E
5
6466



244
G D E E R F F V E
5
6467



254
L S F P D A G F T
5
6468



270
T L L D D S N E D
5
6469



279
F S A S P I F T D
5
6470



303
L P P L E V Y V C
5
6471



307
E V Y V C R V R N
5
6472



314
R N N T C F V D A
5
6473



317
T C F V D A V A E
5
6474



324
A E L A R K A C C
5
6475



330
A G C K L T I C P
5
6476



332
C K L T I C P Q A
5
6477



351
E M E L C Y V Q A
5
6478



352
M E L G Y V Q A P
5
6479



365
P V V F D S P R N
5
6480



400
S V S G L D S F G
5
6481



404
L D S F G N L E V
5
6482



418
A N G K E Y P L G
5
6483



448
D F L H A Q K V Q
5
6484



459
V E L F V D W L A
5
6485



496
C F K L F Q E K Q
5
6486



507
G H G R A L L F Q
5
6487



511
A L L F Q G V V D
5
6488



522
Q V K T I S I N Q
5
6489



533
S N K D L I N Y N
5
6490



550
W N R E V L K R E
5
6491



562
A E C D I I D I P
5
6492



577
R K K A T A F F P
5
6493



590
M L V L G K H L G
5
6494



604
G P I I N G C C C
5
6495



615
E K V R S L L E P
5
6496



628
C T F I D D F T P
5
6497



638
H M L H G E V H C
5
6498



652
R K P F S F K W W
5
6499



11
L E H P T S A V C
4
6500



16
S A V C V A G V E
4
6501



23
V E T L V D I Y G
4
6502



36
G T E M F E V Y G
4
6503



40
F E V Y G T P G V
4
6504



45
T P G V D I Y I S
4
6505



46
P G V D I Y I S P
4
6506



49
D I Y I S P N M E
4
6507



55
N M E R G R E R A
4
6508



76
I I V V N N S P S
4
6509



78
V V M N S P S N D
4
6510



81
N S P S N D L N D
4
6511



85
N D L N D S H V Q
4
6512



98
S S H E P L P L A
4
6513



100
H E P L P L A Y A
4
6514



106
A Y A V L Y L T C
4
6515



112
L T C V D I S L D
4
6516



113
T C V D I S L D C
4
6517



121
C D L N C E G R Q
4
6518



135
D K R Q W V W G P
4
6519



139
W V W C P S G Y G
4
6520



161
C D V Q D N C D Q
4
6521



171
V H C L Q D L E D
4
6522



179
D M S V M V L R T
4
6523



180
M S V M V L R T Q
4
6524



181
S V M V L R T Q G
4
6525



183
M V L R T Q G P A
4
6526



192
A L F D D H K L V
4
6527



196
D H K L V L H T S
4
6528



199
L V L H T S S Y D
4
6529



212
Q V F H T C G P E
4
6530



214
F H I C G P E D V
4
6531



215
H I C G P E D V C
4
6532



224
E A Y R H V L G Q
4
6533



225
A Y R H V L G Q D
4
6534



250
F V E G L S F P D
4
6535



262
T G L I S F H V T
4
6536



268
H V T L L D D S N
4
6537



280
S A S P I F T D T
4
6538



285
F T D T V V F R V
4
6539



296
W I M T P S T L P
4
6540



297
I M T P S T L P P
4
6541



300
P S T L P P L E V
4
6542



308
V Y V C R V R N N
4
6543



309
Y V C R V R N N T
4
6544



312
R V R N N T C F V
4
6545



316
N T C F V D A V A
4
6546



337
C P Q A E N R N D
4
6547



350
D E M E L G Y V Q
4
6548



357
V Q A P H K T L P
4
6549



361
H K T L P V V F D
4
6550



370
S P R N G E L Q D
4
6551



392
V T R E P R D R S
4
6552



403
G L D S F G N L E
4
6553



407
F G N L E V S P P
4
6554



410
L E V S P P V V A
4
6555



411
E V S P P V V A N
4
6556



414
P P V V A N G K E
4
6557



425
L G R I L I G G N
4
6558



429
L I G G N L P G S
4
6559



437
S S G R R V T Q V
4
6560



450
L H A Q K V Q P P
4
6561



457
P P V E L F V D W
4
6562



460
E L F V D W L A V
4
6563



461
L F V D W L A V G
4
6564



464
D W L A V G H V D
4
6565



473
E F L S F V P A P
4
6566



477
F V P A P D G K G
4
6567



483
G K G F R M L L A
4
6568



499
L F Q E K Q K C G
4
6569



513
L F Q C V V D D E
4
6570



539
N Y N K F V Q S C
4
6571



541
N K F V Q S C I D
4
6572



546
S C I D W N R E V
4
6573



568
D I P Q L F K T E
4
6574



584
F P D L V N M L V
4
6575



597
L G I P K P F G P
4
6576



621
L E P L G L H C T
4
6577



625
G L H C T F I D D
4
6578



639
M L H G E V H C G
4
6579



643
E V H C G T N V C
4
6580



2
S L Q R I V R V S
3
6581



6
I V R V S L E H P
3
6582



12
E H P T S A V C V
3
6583



17
A V C V A G V E T
3
6584



20
V A G V E T L V D
3
6585



25
T L V D I Y G S V
3
6586



33
V P E G T E M F E
3
6587



37
T E M F E V Y G T
3
6588



51
Y I S P N M E R G
3
6589



53
S P N M E R G R E
3
6590



73
T L E I I V V M N
3
6591



80
M N S P S N D L N
3
6592



84
S N D L N D S H V
3
6593



110
L Y L T C V D I S
3
6594



119
L D C D L N C E G
3
6595



126
E G R Q D R N F V
3
6596



140
V W G P S G Y G G
3
6597



153
N C D R D D P S C
3
6598



157
D D P S C D V Q D
3
6599



170
H V H C L Q D L E
3
6600



187
T Q G P A A L F D
3
6601



200
V L H T S S Y D A
3
6602



203
T S S Y D A K R A
3
6603



204
S S Y D A K R A Q
3
6604



205
S Y D A K R A Q V
3
6605



217
C G P E D V C E A
3
6606



230
L G Q D K V S Y E
3
6607



237
Y E V P R L H G D
3
6608



251
V E G L S F P D A
3
6609



261
F T G L I S F H V
3
6610



267
F H V T L L D D S
3
6611



273
D D S N S D F S A
3
6612



275
S N E D F S A S P
3
6613



288
T V V F R V A P W
3
6614



293
V A P W I M T P S
3
6615



294
A P W I H T P S T
3
6616



302
T L P P L E V Y V
3
6617



305
P L E V Y V C R V
3
6618



310
V C R V R N N T C
3
6619



339
Q A E N R N D R W
3
6620



395
E P R D R S V S G
3
6621



406
S F G N L E V S P
3
6622



409
N L E V S P P V V
3
6623



416
V V A N G K E Y P
3
6624



424
P L G R I L I G G
3
6625



430
I G G N L P G S S
3
6626



434
L P G S S G R R V
3
6627



435
P G S S G R R V T
3
6628



449
F L H A Q K V Q P
3
6629



452
A Q K V Q P P V E
3
6630



455
V Q P P V E L F V
3
6631



456
Q P P V E L F V D
3
6632



465
W L A V G H V D E
3
6633



470
H V D E F L S F V
3
6634



471
V D E F L S F V P
3
6635



488
M L L A S P G A C
3
6636



501
Q E K Q K C G H G
3
6637



508
H G R A L L F Q G
3
6638



514
F Q G V V D D E Q
3
6639



515
Q G V V D D E Q V
3
6640



525
T I S I N Q V L S
3
6641



531
V L S N K D L I N
3
6642



536
D L I N Y N K F V
3
6643



543
F V Q S C I D W N
3
6644



545
Q S C I D W N R E
3
6645



555
L K R E L G L A E
3
6646



563
E C D I I D I P Q
3
6647



572
L F K T E R K K A
3
6648



586
D L V N M L V L G
3
6649



592
V L G K H L G I P
3
6650



596
H L G I P K P F G
3
6651



600
P K P F G P I I N
3
6652



606
I I N G C C C L E
3
6653



607
I N G C C C L E E
3
6654



620
L L E P L G L H C
3
6655



635
T P Y H M L H G E
3
6656



646
C G T N V C R K P
3
6657



648
T N V C R K P F S
3
6658



656
S F K W W N M V P
3
6659



10
S L E H P T S A V
2
6660



13
H P T S A V C V A
2
6661



14
P T S A V C V A G
2
6662



15
T S A V C V A G V
2
6663



26
L V D I Y C S V P
2
6664



29
I Y G S V P E G T
2
6665



30
Y G S V P E C T E
2
6666



34
P E G T E M F E V
2
6667



39
M F E V Y C T P G
2
6668



56
M E R G R E R A D
2
6669



70
F D A T L E I I V
2
6670



82
S P S N D L N D S
2
6671



91
H V Q I S Y H S S
2
6672



93
Q I S Y H S S H E
2
6673



114
C V D I S L D C D
2
6674



118
S L D C D L N C E
2
6675



122
D L N C E G R Q D
2
6676



132
N F V D K R Q W V
2
6677



150
L L V N C D R D D
2
6678



160
S C D V Q D N C D
2
6679



168
D Q H V H C L Q D
2
6680



176
D L E D M S V M V
2
6681



182
V M V L R T Q G P
2
6682



213
V F H I C G P E D
2
6683



232
Q D K V S Y E v P
2
6684



236
S Y E V P R L H G
2
6685



239
V P R L H G D E E
2
6686



243
H G D E F R F F V
2
6687



245
D E E R F F V E G
2
6688



252
E G L S F P D A G
2
6689



266
S F H V T L L D D
2
6690



274
D S N F D F S A S
2
6691



278
D F S A S P I F T
2
6692



281
A S P I F T D T V
2
6693



286
T D T V V F R V A
2
6694



287
D T V V F R V A P
2
6695



299
T P S T L P P L E
2
6696



319
F V D A V A E L A
2
6697



333
K L T I C P Q A E
2
6698



349
Q D F M E L G Y V
2
6699



358
Q A P H K T L P V
2
6700



363
T L P V V F D S P
2
6701



367
V F D S P R N C E
2
6702



378
D F P Y K R I L G
2
6703



380
P Y K R I L C P D
2
6704



384
I L G P D F G Y V
2
6705



385
L G P D F G Y V T
2
6706



388
D F C Y V T R E P
2
6707



401
V S G L D S F C N
2
6708



438
S G R R V T Q V V
2
6709



444
Q V V R D F L H A
2
6710



463
V D W L A V G H V
2
6711



474
F L S F V P A P D
2
6712



493
P G A C F K L F Q
2
6713



537
L I N Y N K F V Q
2
6714



554
V L K R E L G L A
2
6715



573
F K T E R K K A T
2
6716



578
K K A T A F F P D
2
6717



603
F G P I I N G C C
2
6718



614
E E K V R S L L E
2
6719



624
L G L H C T F I D
2
6720



627
H C T F I D D F T
2
6721



636
P Y H M L H G E V
2
6722



640
L H G E V H C G T
2
6723



63
A D T R R W R F D
1
6724



87
L N D S H V Q I S
1
6725



96
Y H S S H E P L P
1
6726



116
D I S L D C D L N
1
6727



134
V D K R Q W V W G
1
6728



152
V N C D R D D P S
1
6729



154
C D R D D P S C D
1
6730



158
D P S C D V Q D N
1
6731



163
V Q D N C D Q H V
1
6732



165
D N C D Q H V H C
1
6733



167
C D Q H V H C L Q
1
6734



173
C L Q D L E D M S
1
6735



184
V L R T Q G P A A
1
6736



188
Q G P A A L F D D
1
6737



221
D V C E A Y R H V
1
6738



223
C E A Y R H V L G
1
6739



238
E V P R L H G D E
1
6740



255
S F P D A G F T G
1
6741



258
D A G F T G L I S
1
6742



315
N N T C F V D A V
1
6743



323
V A E L A R K A G
1
6744



341
E N R N D R W I Q
1
6745



347
W I Q D E M E L G
1
6746



445
V V R D F L H A Q
1
6747



580
A T A F F P D L V
1
6748



634
F T P Y H N L H G
1
6749



641
H G E V H C C T N
1
6750











184P1E2 v.2: HLA Peptide


Scoring Results B2705 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







9
A P L E V Y V C R
16
6751



6
S T L A P L E V Y
13
6752



3
M T P S T L A P L
12
6753



8
L A P L E V Y V C
5
6754



1
W I M T P S T L A
4
6755



2
I M T P S T L A P
4
6756



4
T P S T L A P L E
3
6757



5
P S T L A P L E V
3
6758



7
T L A P L E V Y V
3
6759











184P1E2 v.3: HLA Peptide


Scoring Results B2705 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.







5
S F V P V P D G K
13
6760



8
P V P D G K G F R
13
6761



9
V P D G K G F R M
13
6762



7
V P V P D G K G F
12
6763



1
D E F L S F V P V
6
6764



4
L S F V P V P D G
6
6765



2
E F L S F V P V P
5
6766



6
F V P V P D G K G
4
6767



3
F L S F V P V P D
2
6768

















TABLE XXXII







184P1E2 v.1: HLA Peptide


Scoring Results B2709 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






426
G R I L I G G N L
26
6769



66
R R W R F D A T L
24
6770



68
W R F D A T L E I
21
6771



185
L R T Q G P A A L
21
6772



396
P R D R S V S G L
21
6773



509
G R A L L F Q G V
21
6774



551
N R E V L K R E L
20
6775



311
C R V R N N T C F
19
6776



393
T R E P R D R S V
19
6777



155
D R D D P S C D V
18
6778



371
P R N G E L Q D F
18
6779



440
R R V T Q V V R D
18
6780



576
E R K K A T A F F
18
6781



618
R S L L E P L G L
17
6782



263
G L I S F H V T L
16
6783



346
R W I Q D E M E L
16
6784



234
K V S Y E V P R L
15
6785



248
R F F V E G L S F
15
6786



421
K E Y P L G R I L
15
6787



447
R D F L H A Q K V
15
6788



524
K T I S I N Q V L
15
6789



582
A F F P D L V N M
15
6790



59
G R E R A D T R R
14
6791



191
A A L F D D H K L
14
6792



241
R L H G D E E R F
14
6793



399
R S V S G L D S F
14
6794



408
G N L E V S P P V
14
6795



439
G R R V T Q V V R
14
6796



441
R V T Q V V R D F
14
6797



510
R A L L F Q G V V
14
6798



564
C D I I D I P Q L
14
6799



579
K A T A F F P D L
14
6800



31
G S V P E G T E M
13
6801



62
R A D T R R W R F
13
6802



72
A T L E I I V V M
13
6803



101
E P L P L A Y A V
13
6804



127
G R Q D R N F V D
13
6805



142
G P S G Y G G I L
13
6806



186
R T Q G P A A L F
13
6807



227
R H V L G Q D K V
13
6808



328
R K A G C K L T I
13
6809



377
Q D F P Y K R I L
13
6810



382
K R I L G P D F G
13
6811



402
S G L D S F C N L
13
6812



469
G H V D E F L S F
13
6813



553
E V L K R E L G L
13
6814



585
P D L V N M L V L
13
6815



595
K H L C I P K P F
13
6816



616
K V R S L L E P L
13
6817



642
G H V H C G T N V
13
6818



1
M S L Q R I V R V
12
6819



4
Q R I V R V S L E
12
6820



7
V R V S L H H P T
12
6821



18
V C V A G V E T L
12
6822



48
V D I Y I S P N N
12
6823



69
R F D A T L H I I
12
6824



104
P L A Y A V L Y L
12
6825



115
V D I S L D C D L
12
6826



136
K R Q W V W G P S
12
6827



192
A L F D D H K L V
12
6828



209
K R A Q V F H I C
12
6829



231
G Q D K V S Y E V
12
6830



240
P R L H G D E H R
12
6831



247
E R F F V H C L S
12
6832



277
H D F S A S P I F
12
6833



283
P I F T D T V V F
12
6834



289
V V F R V A P W I
12
6835



298
M T P S T L P P L
12
6836



312
R V R N N T C F V
12
6837



318
C F V D A V A E L
12
6838



426
G K E Y P L G R I
12
6839



453
Q K V Q P P V E L
12
6840



454
K V Q P P V E L F
12
6841



460
E L F V D W L A V
12
6842



505
K C G H G R A L L
12
6843



535
K D L I N Y N K F
12
6844



556
K R E L G L A E C
12
6845



589
N M L V L G K H L
12
6846



632
D D F T P Y H M L
12
6847



3
L Q R I V R V S L
11
6848



21
A G V E T L V D I
11
6849



44
G T P G V D I Y I
11
6850



79
V M N S P S N D L
11
6851



95
S Y H S S H E P L
11
6852



97
H S S H E P L P L
11
6853



109
V L Y L T C V D I
11
6854



111
Y L T C V D I S L
11
6855



130
D R N F V D K R Q
11
6856



144
S C Y C C I L L V
11
6857



166
N C D Q H V H C L
11
6858



169
Q H V H C L Q D L
11
6859



172
H C L Q D L E D M
11
6860



175
Q D L E D M S V M
11
6861



177
L E D M S V M V L
11
6862



193
L F D D H K L V L
11
6863



246
E E R F F V E G L
11
6864



253
C L S F P D A G F
11
6865



259
A G F T G L I S F
11
6866



264
L I S F H V T L L
11
6867



291
F R V A P W I M T
11
6868



295
P W I M T P S T L
11
6869



327
A R K A G C K L T
11
6870



342
N R N D R W I Q D
11
6871



368
F D S P R N C E L
11
6872



384
I L G P D F G Y V
11
6873



398
D R S V S C L D S
11
6874



417
V A N C K E Y P L
11
6875



446
V R D F L H A Q K
11
6876



458
P V E L F V D W L
11
6877



467
A V C H V D E F L
11
6878



486
F R M L L A S P G
11
6879



491
A S P G A C F K L
11
6880



515
Q G V V D D E Q V
11
6881



529
N Q V L S N K D L
11
6882



530
Q V L S N K D L I
11
6883



583
F F P D L V N M L
11
6884



591
L V L G K H L G I
11
6885



598
G I P K P F G P I
11
6886



599
I P K P F G P I I
11
6887



605
P I I N G C C C L
11
6888



609
G C C C L E E K V
11
6889



612
C L E E K V R S L
11
6890



617
V R S L L E P L G
11
6891



647
G T N V C R K P F
11
6892



651
C R K P F S F K W
11
6893



12
E H P T S A V C V
10
6894



25
T L V D I Y G S V
10
6895



40
F E V Y G T P G V
10
6896



57
E R G R E R A D T
10
6897



61
E R A D T R R W R
10
6898



65
T R R W R F D A T
10
6899



86
D L N D S H V Q I
10
6900



102
P L P L A Y A V L
10
6901



143
P S G Y G G I L L
10
6902



208
A K R A Q V F H I
10
6903



222
V C E A Y R H V L
10
6904



226
Y R H V L G Q D K
10
6905



256
F P D A G F T G L
10
6906



285
F T D T V V F R V
10
6907



300
P S T L P P L E V
10
6908



302
T L P P L E V Y V
10
6909



305
P L E V Y V C R V
10
6910



313
V R N N T C F V D
10
6911



315
N N T C F V D A V
10
6912



326
L A R K A G C K L
10
6913



345
D R W I Q D E M H
10
6914



356
Y V Q A P H K T L
10
6915



359
A P H K T L P V V
10
6916



434
L P C S S G R R V
10
6917



437
S S C R R V T Q V
10
6918



442
V T Q V V R D F L
10
6919



455
V Q P P V F L F V
10
6920



466
L A V C H V D E F
10
6921



478
V P A P D G K G F
10
6922



480
A P D G K G F R M
10
6923



481
P D C K G F R M L
10
6924



482
D C K G F R M L L
10
6925



504
Q K C G H G R A L
10
6926



506
C G H G R A L L F
10
6927



523
V K T I S I N Q V
10
6928



547
C I D W N R E V L
10
6929



559
L G L A E C D I I
10
6930



565
D I I D I P Q L F
10
6931



613
L E E K V R S L L
10
6932



622
E P L G L H C T F
10
6933



631
I D D F T P Y H M
10
6934



655
F S F K W W N M V
10
6935



15
T S A V C V A G V
9
6936



42
V Y G T P G V D I
9
6937



71
D A T L E I I V V
9
6938



107
Y A V L Y L T C V
9
6939



125
C E G R Q D R N F
9
6940



126
E G R Q D R N F V
9
6941



132
N F V D K R Q W V
9
6942



141
W G P S G Y C G I
9
6943



176
D L S D M S V M V
9
6944



205
S Y D A K R A Q V
9
6945



206
Y D A K R A Q V F
9
6946



214
F H I C G P F D V
9
6947



261
F T C L I S F H V
9
6948



276
N E D F S A S P I
9
6949



281
A S P I F T D T V
9
6950



282
S P I F T D T V V
9
6951



340
A E N R N D R W I
9
6952



344
N D R W I Q D E M
9
6953



358
Q A P H K T L P V
9
6954



360
P H K T L P V V F
9
6955



376
L Q D F P Y K R I
9
6956



381
Y K R I L G P D F
9
6957



404
L D S F G N L E V
9
6958



409
N L E V S P P V V
9
6959



451
H A Q K V Q P P V
9
6960



463
V D W L A V G H V
9
6961



489
L L A S P G A C F
9
6962



518
V D D B Q V K T I
9
6963



536
D L I N Y N K F V
9
6964



546
S C I D W N R E V
9
6965



561
L A E C D I I D I
9
6966



575
T B R K K A T A F
9
6967



580
A T A F F P D L V
9
6968



654
P F S F K W W N M
9
6969



10
S L B H P T S A V
8
6970



19
C V A G V E T L V
8
6971



32
S V P E G T E M F
8
6972



34
P E G T E M F E V
8
6973



70
F D A T L E I I V
8
6974



84
S N D L N D S H V
8
6975



131
R N F V D K R Q W
8
6976



163
V Q D N C D Q H V
8
6977



174
L Q D L E D M S V
8
6978



221
D V C E A Y R H V
8
6979



242
L H G D E E R F F
8
6980



243
H C D F E R F F V
8
6981



257
P D A G F T G L I
8
6982



271
L L D D S N E D F
8
6983



290
V F R V A P W I M
8
6984



349
Q D E M E L G Y V
8
6985



422
E Y P L G R I L I
8
6986



438
S G R R V T Q V V
8
6987



470
H V D E F L S F V
8
6988



492
S P G A C F K L F
8
6989



520
D B Q V K T I S I
8
6990



540
Y N K F V Q S C I
8
6991



558
E L G L A E C D I
8
6992



584
F P D L V N M L V
8
6993



623
P L G L H C T F I
8
6994



626
L H C T F I D D F
8
6995



636
P Y H M L H G E V
8
6996



649
N V C R K P F S F
8
6997



374
G E L Q D F P Y K
7
6998



487
R M L L A S P G A
7
6999



557
R E L G L A E C D
7
7000



5
R I V R V S L E H
6
7001



60
R E R A D T R R W
6
7002



67
R W R F D A T L E
6
7003



362
K T L P V V F D S
6
7004



383
R I L G P D F G Y
6
7005



390
C Y V T R E P R D
6
7006



427
R I L I G G N L P
6
7007



432
G N L P G S S G R
6
7008



8
R V S L E H P T S
5
7009



58
R G R B R A D T R
5
7010



117
I S L D C D L N C
5
7011



128
R Q D R N F V D K
5
7012



137
R Q W V W G P S G
5
7013



145
C Y G G I L L V N
5
7014



147
G G I L L V N C D
5
7015



148
G I L L V N C D R
5
7016



156
R D D P S C D V Q
5
7017



254
L S F P D A G F T
5
7018



292
R V A P W I M T P
5
7019



314
R N N T C F V D A
5
7020



317
T C F V D A V A F
5
7021



332
C K L T I C P Q A
5
7022



394
R F P R D R S V S
5
7023



397
R D R S V S G L D
5
7024



473
F F L S F V P A P
5
7025



48
K G F R M L L A S
5
7026



516
G V V D D F Q V K
5
7027



538
I N Y N K F V Q S
5
7028



552
R E V L K R E L G
5
7029



577
R K K A T A F F P
5
7030



601
K P F C P I I N C
5
7031



653
K P F S F K W W N
5
7032



22
G V E T L V D I Y
4
7033



24
E T L V D I Y G S
4
7034



28
D I Y G S V P E G
4
7035



36
G T E M F E V Y G
4
7036



103
L P L A Y A V L Y
4
7037



149
I L L V N C D R D
4
7038



210
R A Q V F H I C G
4
7039



220
E D V C E A Y R H
4
7040



224
E A Y R H V L G Q
4
7041



260
C F T C L I S F H
4
7042



265
I S F H V T L L D
4
7043



269
V T L L D D S N E
4
7044



284
I F T D T V V F R
4
7045



297
I N T P S T L P P
4
7046



307
E V Y V C R V R N
4
7047



343
R N D R W I Q D E
4
7048



354
L C Y V Q A P H K
4
7049



355
G Y V Q A P H K T
4
7050



365
P V V F D S P R N
4
7051



372
R N G E L Q D F P
4
7052



386
G P D F C Y V T R
4
7053



387
P D F G Y V T R F
4
7054



403
C L D S F C N L E
4
7055



436
C S S G R R V T Q
4
7056



475
L S F V P A P D G
4
7057



483
G K G F R M L L A
4
7058



494
G A C F K L F Q E
4
7059



498
K L F Q F K Q K C
4
7060



507
G H C R A L L F Q
4
7061



511
A L L F Q C V V D
4
7062



512
L L F Q G V V D D
4
7063



526
I S I N Q V L S N
4
7064



594
G K H L C I P K P
4
7065



604
G P I I N G C C C
4
7066



611
C C L F E K V R S
4
7067



652
R K P F S F K W W
4
7068



13
H P T S A V C V A
3
7069



43
Y G T P G V D I Y
3
7070



47
G V D I Y I S P N
3
7071



50
I Y I S P N M F R
3
7072



73
T L E I I V V M N
3
7073



74
L E I I V V M N S
3
7074



77
I V V M N S P S N
3
7075



81
N S P S N D L N D
3
7076



85
N D L N D S H V Q
3
7077



90
S H V Q I S Y H S
3
7078



94
I S Y H S S H E P
3
7079



105
L A Y A V L Y L T
3
7080



108
A V L Y L T C V D
3
7081



110
L Y L T C V D I S
3
7082



113
T C V D I S L D C
3
7083



121
C D L N C E G R Q
3
7084



157
D D P S C D V Q D
3
7085



179
D M S V M V L R T
3
7086



187
T Q G P A A L F D
3
7087



189
G P A A L F D D H
3
7088



198
K L V L H T S S Y
3
7089



203
T S S Y D A K R A
3
7090



212
Q V F H I C G P E
3
7091



216
I C C P F D V C F
3
7092



218
G P F D V C E A Y
3
7093



233
D K V S Y F V P R
3
7094



244
C D E E R F F V E
3
7095



252
F C L S F P D A C
3
7096



270
T L L D D S N E D
3
7097



301
S T L P P L E V Y
3
7098



304
P P L E V Y V C R
3
7099



308
V Y V C R V R N N
3
7100



324
A E L A R K A G C
3
7101



331
G C K L T I C P Q
3
7102



333
K L T I C P Q A F
3
7103



335
T I C P Q A F N R
3
7104



352
M F L G Y V Q A P
3
7105



361
H K T L P V V F D
3
7106



366
V V F D S P R N C
3
7107



379
F P Y K R I L G P
3
7108



410
L F V S P P V V A
3
7109



411
F V S P P V V A N
3
7110



412
V S P P V V A N G
3
7111



423
Y P L G R I L I G
3
7112



428
I L I G G N L P G
3
7113



431
G G N L P G S S G
3
7114



444
Q V V R D F L H A
3
7115



472
D E F L S F V P A
3
7116



485
G F R M L L A S P
3
7117



495
A C F K L F Q E K
3
7118



497
F K L F Q E K Q K
3
7119



503
K Q K C G H G R A
3
7120



534
N K D L I N Y N K
3
7121



542
K F V Q S C I D W
3
7122



560
G L A E C D I I D
3
7123



567
I D I P Q L F K T
3
7124



570
P Q L F K T E R K
3
7125



581
T A F F P D L V N
3
7126



625
G L H C T F I D D
3
7127



628
C T F I D D F T P
3
7128



638
H M L H G E V H C
3
7129



2
S L Q R I V R V S
2
7130



9
V S L E H P T S A
2
7131



16
S A V C V A G V E
2
7132



17
A V C V A G V E T
2
7133



27
V D I Y G S V P E
2
7134



29
I Y G S V P E G T
2
7135



37
T E M F E V Y G T
2
7136



38
E M F E V Y G T P
2
7137



41
E V Y G T P G V D
2
7138



45
T P G V D I Y I S
2
7139



46
P G V D I Y I S P
2
7140



49
D I Y I S P N M E
2
7141



51
Y I S P N N E R G
2
7142



63
A D T R R W R F D
2
7143



76
I I V V N N S P S
2
7144



87
L N D S H V Q I S
2
7145



92
V Q I S Y H S S H
2
7146



96
Y H S S H E P L P
2
7147



99
S H S P L P L A Y
2
7148



106
A Y A V L Y L T C
2
7149



140
V W G P S C Y G C
2
7150



146
Y G G I L L V N C
2
7151



153
N C D R D D P S C
2
7152



158
D P S C D V Q D N
2
7153



161
C D V Q D N C D Q
2
7154



168
D Q H V H C L Q D
2
7155



178
E D M S V M V L R
2
7156



183
M V L R T Q G P A
2
7157



195
D D H K L V L H T
2
7158



197
H K L V L H T S S
2
7159



199
L V L H T S S Y D
2
7160



201
L H T S S Y D A K
2
7161



204
S S Y D A K R A Q
2
7162



211
A Q V F H I C G P
2
7163



217
C G P E D V C E A
2
7164



225
A Y R H V L G Q D
2
7165



228
H V L G Q D K V S
2
7166



229
V L G Q D K V S Y
2
7167



235
V S Y E V P R L H
2
7168



237
Y E V P R L H G D
2
7169



262
T G L I S F H V T
2
7170



280
S A S P I F T D T
2
7171



287
D T V V F R V A P
2
7172



288
T V V F R V A P W
2
7173



294
A P W I M T P S T
2
7174



303
L P P L E V Y V C
2
7175



321
D A V A E L A R K
2
7176



322
A V A E L A R K A
2
7177



329
K A G C K L T I C
2
7178



336
I C P Q A E N R N
2
7179



348
I Q D E M E L G Y
2
7180



351
E M E L G Y V Q A
2
7181



370
S P R N G E L Q D
2
7182



389
F G Y V T R E P R
2
7183



405
D S F G N L E V S
2
7184



418
A N G K S Y P L G
2
7185



448
D F L H A Q K V Q
2
7186



449
F L H A Q K V Q P
2
7187



452
A Q K V Q P P V E
2
7188



457
P P V S L F V D W
2
7189



459
V E L F V D W L A
2
7190



462
F V D W L A V G H
2
7191



464
D W L A V G H V D
2
7192



476
S F V P A P D G K
2
7193



488
M L L A S P G A C
2
7194



490
L A S P G A C F K
2
7195



522
Q V K T I S I N Q
2
7196



532
L S N K D L I N Y
2
7197



541
N K F V Q S C I D
2
7198



544
V Q S C I D W N R
2
7199



545
Q S C I D V W R F
2
7200



548
I D W N R E V L K
2
7201



549
D W N R E V L K R
2
7202



562
A E C D I I D I P
2
7203



566
I I D I P Q L F K
2
7204



571
Q L F K T E R K K
2
7205



574
K T E R K K A T A
2
7206



578
K K A T A F F P D
2
7207



586
D L V N M L V L G
2
7208



587
L V N M L V L G K
2
7209



597
L G I P K P F G P
2
7210



602
P F G P I I N G C
2
7211



607
I N G C C C L E E
2
7212



615
E K V R S L L E P
2
7213



619
S L L E P L G L H
2
7214



620
L L E P L G L H C
2
7215



624
L G L H C T F I D
2
7216



627
H C T F I D D F T
2
7217



629
T F I D D F T P Y
2
7218



633
D F T P Y H M L H
2
7219



635
T P Y H M L H G E
2
7220



645
H C G T N V C R K
2
7221



648
T N V C R K P F S
2
7222



6
I V R V S L E H P
1
7223



11
L E H P T S A V C
1
7224



14
P T S A V C V A G
1
7225



20
V A G V E T L V D
1
7226



23
V E T L V D I Y G
1
7227



35
E C T E M F E V Y
1
7228



52
I S P N M E R G R
1
7229



53
S P N M E R G R E
1
7230



55
N M E R G R E R A
1
7231



75
E I I V V M N S P
1
7232



78
V V M N S P S N D
1
7233



80
M N S P S N D L N
1
7234



82
S P S N D L N D S
1
7235



98
S S H E P L P L A
1
7236



112
L T C V D I S L D
1
7237



116
D I S L D C D L N
1
7238



118
S L D C D L N C E
1
7239



122
D L N C E C R Q D
1
7240



124
N C E G R Q D R N
1
7241



129
Q D R N F V D K R
1
7242



134
V D K R Q W V W G
1
7243



135
D K R Q W V W G P
1
7244



138
Q W V W G P S G Y
1
7245



150
L L V N C D R D D
1
7246



151
L V N C D R D D P
1
7247



154
C D R D D P S C D
1
7248



159
P S C D V Q D N C
1
7249



162
D V Q D N C D Q H
1
7250



164
Q D N C D Q H V H
1
7251



165
D N C D Q H V H C
1
7252



171
V H C L Q D L E D
1
7253



180
M S V M V L R T Q
1
7254



181
S V M V L R T Q G
1
7255



182
V M V L R T Q G P
1
7256



184
V L R T Q G P A A
1
7257



194
F D D H K L V L H
1
7258



200
V L H T S S Y D A
1
7259



202
H T S S Y D A K R
1
7260



207
D A K R A Q V F H
1
7261



213
V F H I C C P 5 D
1
7262



223
C E A Y R H V L G
1
7263



230
L G Q D K V S Y E
1
7264



236
S Y E V P R L H G
1
7265



245
D E E R F F V E G
1
7266



249
F F V E G L S F P
1
7267



251
V E G L S F P D A
1
7268



258
D A G F T G L I S
1
7269



266
S F H V T L L D D
1
7270



267
F H V T L L D D S
1
7271



268
H V T L L D D S N
1
7272



272
L D D S N E D F S
1
7273



274
D S N E D F S A S
1
7274



279
F S A S P I F T D
1
7275



286
T D T V V F R V A
1
7276



293
V A P W I M T P S
1
7277



299
T P S T L P P L E
1
7278



306
L E V Y V C R V R
1
7279



309
Y V C R V R N N T
1
7280



319
F V D A V A E L A
1
7281



320
V D A V A E L A R
1
7282



325
E L A R K A G C K
1
7283



330
A G C K L T I C P
1
7284



334
L T I C P Q A E N
1
7285



337
C P Q A E N R N D
1
7286



339
Q A E N R N D R W
1
7287



341
E N R N D R W I Q
1
7288



350
D E M E L G Y V Q
1
7289



357
V Q A P H K T L P
1
7290



364
L P V V F D S P R
1
7291



367
V F D S P R N G E
1
7292



375
E L Q D F P Y K R
1
7293



380
P Y K R I L C P D
1
7294



385
L G P D F G Y V T
1
7295



391
Y V T R E P R D R
1
7296



392
V T R E P R D R S
1
7297



395
E P R D R S V S G
1
7298



401
V S G L D S F G N
1
7299



406
S F G N L E V S P
1
7300



407
F G N L E V S P P
1
7301



413
S P P V V A N G K
1
7302



414
P P V V A N G K E
1
7303



415
P V V A N G K E Y
1
7304



419
N G K E Y P L G R
1
7305



424
P L G R I L I G G
1
7306



429
L I G G N L P G S
1
7307



430
I G G N L P G S S
1
7308



433
N L P G S S G R R
1
7309



435
P G S S G R R V T
1
7310



443
T Q V V R D F L H
1
7311



450
L H A Q K V Q P P
1
7312



461
L F V D W L A V G
1
7313



465
W L A V G H V D E
1
7314



468
V G H V D E F L S
1
7315



493
P G A C F K L F Q
1
7316



508
H G R A L L F Q G
1
7317



513
L F Q G V V D D E
1
7318



517
V V D D E Q V K T
1
7319



521
E Q V K T I S I N
1
7320



525
T I S I N Q V L S
1
7321



527
S I N Q V L S N K
1
7322



528
I N Q V L S N K D
1
7323



531
V L S N K D L I N
1
7324



539
N Y N K F V Q S C
1
7325



543
F V Q S C I D W N
1
7326



550
W N R E V L K R E
1
7327



555
L K R E L G L A E
1
7328



563
E C D I I D I P Q
1
7329



569
I P Q L F K T E R
1
7330



573
F K T E R K K A T
1
7331



590
M L V L G K H L G
1
7332



596
H L G I P K P F G
1
7333



603
F G P I I N G C C
1
7334



606
I I N G C C C L E
1
7335



614
E E K V R S L L E
1
7336



630
F I D D F T P Y H
1
7337



634
F T P Y H M L H G
1
7338



639
M L H G E V H C G
1
7339



644
V H C G T N V C R
1
7340



646
C G T N V C R K P
1
7341










184P1E2 v.2: HLA Peptide


Scoring Results B2709 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






3
M T P S T L A P L
11
7342



5
P S T L A P L E V
10
7343



7
T L A P L E V Y V
10
7344



2
I M T P S T L A P
4
7345



9
A P L E V Y V C R
4
7346



4
T P S T L A P L E
2
7347



6
S T L A P L E V Y
2
7348



8
L A P L E V Y V C
2
7349










184P1E2 v.3: HLA Peptide


Scoring Results B2709 9-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID NO.






1
D E F L S F V P V
11
7350



7
V P V P D G K G F
11
7351



9
V P D G K G F R M
9
7352



2
E F L S F V P V P
5
7353



4
L S F V P V P D G
4
7354



5
S F V P V P D G K
2
7355



3
F L S F V P V P D
1
7356



















TABLE XXXIII








SEQ





ID


Pos
1 2 3 4 5 6 7 8 9
score
NO.















184P1E2 v.1: HLA Peptide Scoring Results B4402 9-


mers SYFPEITHI










421
K E Y P L G R I L
25
7357


340
A E N R N D R W I
24
7358


177
L E D M S V M V L
23
7359


246
E E R F F V E G L
23
7360


575
T E R K K A T A F
23
7361


60
R E R A D T R R W
22
7362


125
C E G R Q D R N F
22
7363


613
L E E K V R S L L
22
7364


276
N E D F S A S P I
21
7365


520
D E Q V K T I S I
20
7366


99
S H E P L P L A Y
18
7367


259
A G F T G L I S F
18
7368


562
A E C D I I D I P
18
7369


301
S T L P P L E V Y
17
7370


564
C D I I D I P Q L
17
7371


565
D I I D I P Q L F
17
7372


595
K H L G I P K P F
17
7373


191
A A L F D D H K L
16
7374


237
Y E V P R L H G D
16
7375


283
P I F T D T V V F
16
7376


377
Q D F P Y K R I L
16
7377


454
K V Q P P V E L F
16
7378


491
A S P G A C F K L
16
7379


52
K T I S I N Q V L
16
7380


56
M E R G R E R A D
15
7381


263
G L I S F H V T L
15
7382


295
P W I M T P S T L
15
7383


324
A E L A R K A G C
15
7384


352
M E L G Y V Q A P
15
7385


368
F D S P R N G E L
15
7386


422
E Y P L G R I L I
15
7387


426
G R I L I G G N L
15
7388


472
D E F L S F V P A
15
7389


478
V P A P D G K G F
15
7390


492
S P G A C F K L F
15
7391


506
C G H G R A L L F
15
7392


535
K D L I N Y N K F
15
7393


553
E V L K R E L G L
15
7394


614
E E K V R S L L E
15
7395


622
E P L G L H C T F
15
7396


629
T F I D D F T P Y
15
7397


11
L E H P T S A V C
14
7398


35
E G T E M F E V Y
14
7399


43
Y G T P C V D I Y
14
7400


62
R A D T R R W R F
14
7401


74
L E I I V V M N S
14
7402


103
L P L A Y A V L Y
14
7403


131
R N F V D K R Q W
14
7404


166
N C D Q H V H C L
14
7405


193
L F D D H K L V L
14
7406


277
E D F S A S P I F
14
7407


360
P H K T L P V V F
14
7408


394
R E P R D R S V S
14
7409


402
S G L D S F G N L
14
7410


504
Q K C G H G R A L
14
7411


529
N O V L S N K D L
14
7412


547
C I D W N R E V L
14
7413


583
F F P D L V N M L
14
7414


585
P D L V N M L V L
14
7415


589
N M L V L G K H L
14
7416


605
P I I N G C C C L
14
7417


618
R S L L E P L G L
14
7418


621
L E P L C L H C T
14
7419


626
L H C T F I D D F
14
7420


632
D D F T P Y H M L
14
7421


651
C R K P F S F K W
14
7422


652
R K P F S F K W W
14
7423


21
A G V S T L V D I
13
7424


32
S V P E G T E M F
13
7425


68
W R F D A T L E I
13
7426


88
N D S H V Q I S Y
13
7427


97
H S S H E P L P L
13
7428


100
H E P L P L A Y A
13
7429


102
P L P L A Y A V L
13
7430


115
V D I S L D C D L
13
7431


133
F V D K R Q W V W
13
7432


185
L R T Q G P A A L
13
7433


186
R T Q G P A A L F
13
7434


198
K L V L H T S S Y
13
7435


206
Y D A K R A Q V F
13
7436


218
G P E D V C E A Y
13
7437


222
V C E A Y R H V L
13
7438


223
C E A Y R H V L G
13
7439


234
K V S Y E V P R L
13
7440


245
D E E R F F V E G
13
7441


248
R F F V E C L S F
13
7442


264
L I S F H V T L L
13
7443


346
R W I Q D E M E L
13
7444


356
Y V Q A P H K T L
13
7445


396
P R D R S V S G L
13
7446


410
L E V S P P V V A
13
7447


415
P V V A N G K H Y
13
7448


467
A V G H V D E F L
13
7449


469
G H V D E F L S F
13
7450


482
D G K G F R M L L
13
7451


505
K C G H G R A L L
13
7452


518
V D D H Q V K T I
13
7453


552
R E V L K R E L G
13
7454


557
R E L C L A H C D
13
7455


576
E R K K A T A F F
13
7456


647
G T N V C R K P F
13
7457


649
N V C R K P F S F
13
7458


3
L Q R I V R V S L
12
7459


18
V C V A G V E T L
12
7460


22
G V E T L V D I Y
12
7461


23
V E T L V D I Y G
12
7462


37
T E H F E V Y G T
12
7463


66
R R W R F D A T L
12
7464


104
P L A Y A V L Y L
12
7465


142
G P S G Y G G I L
12
7466


143
P S G Y G G I L L
12
7467


208
A K R A Q V F H I
12
7468


242
L H G D E E R F F
12
7469


251
V E G L S F P D A
12
7470


256
F P D A G F T G L
12
7471


271
L L D D S N E D F
12
7472


288
T V V F R V A P W
12
7473


289
V V F R V A P W I
12
7474


298
M T P S T L P P L
12
7475


306
L E V Y V C R V R
12
7476


311
C R V R N N T C F
12
7477


318
C F V D A V A E L
12
7478


339
Q A E N R N D R W
12
7479


348
I Q D E M E L G Y
12
7480


350
D E M E L G Y V Q
12
7481


371
P R N G E L Q D F
12
7482


373
N G E L Q D F P Y
12
7483


37
G E L Q D F P Y K
12
7484


383
R I L G P D F G Y
12
7485


441
R V T Q V V R D F
12
7486


453
Q K V Q P P V E L
12
7487


457
P P V E L F V D W
12
7488


458
P V F L F V D W L
12
7489


459
V E L F V D W L A
12
7490


466
L A V G H V D E F
12
7491


481
P D G K G F R M L
12
7492


532
L S N K D L I N Y
12
7493


542
K F V Q S C I D W
12
7494


551
N R E V L K R E L
12
7495


561
L A E C D I I D I
12
7496


579
K A T A F F P D L
12
7497


612
C L F F K V R S L
12
7498


616
K V R S L L F P L
12
7499


34
P E C T E M F E V
11
7500


40
F E V Y G T P G V
11
7501


44
G T P C V D I Y I
11
7502


79
V M N S P S N D L
11
7503


86
D L N D S H V Q I
11
7504


95
S Y H S S H F P L
11
7505


111
Y L T C V D I S L
11
7506


138
Q W V W C P S G Y
11
7507


169
Q H V H C L Q D L
11
7508


219
P E D V C E A Y R
11
7509


229
V L G Q D K V S Y
11
7510


241
R L H C D E F R F
11
7511


253
G L S F P D A G F
11
7512


257
P D A G F T G L I
11
7513


326
L A R K A G C K L
11
7514


328
R K A G C K L T I
11
7515


376
L Q D F P Y K R I
11
7516


399
R S V S G L D S F
11
7517


417
V A N G K E Y P L
11
7518


442
V T Q V V R D F L
11
7519


530
Q V L S N K D L I
11
7520


558
E L G L A E C D I
11
7521


591
L V L G K H L G I
11
7522


599
I P K P F G P I I
11
7523


42
V Y G T P G V D I
10
7524


69
R F D A T L E I I
10
7525


72
A T L E I I V V M
10
7526


109
V L Y L T C V D I
10
7527


381
Y K R I L G P D F
10
7528


411
E V S P P V V A N
10
7529


489
L L A S P C A C F
10
7530


501
Q E K Q K C G H G
10
7531


582
A F F P D L V N M
10
7532


598
G I P K P F C P I
10
7533


642
G E V H C G T N V
10
7534


101
S P L P L A Y A V
9
7535


141
W G P S G Y G G I
9
7536


192
A L F D D H K L V
9
7537


420
C K F Y P L G R I
9
7538


559
L G L A E C D I I
9
7539


623
P L C L H C T F I
9
7540


484
K G F R M L L A S
8
7541


540
Y N K F V Q S C I
8
7542


567
I D I P Q L F K T
8
7543


601
K P F C P I I N G
8
7544


602
P F G P I I N G C
8
7545


144
S G Y C C I L L V
7
7546


280
S A S P I F T D T
7
7547


292
R V A P W I M T P
7
7548


423
Y P L C R I L I G
7
7549


460
F L F V D W L A V
7
7550


597
L G I P K P F C P
7
7551


1
M S L Q R I V R V
6
7552


2
S L Q R I V R V S
6
7553


4
Q R I V R V S L E
6
7554


12
E H P T S A V C V
6
7555


24
E T L V D I Y G S
6
7556


63
A D T R R W R F D
6
7557


71
D A T L E I I V V
6
7558


75
E I I V V M N S P
6
7559


108
A V L Y L T C V D
6
7560


147
G G I L L V N C D
6
7561


211
A Q V F H I C C P
6
7562


252
E G L S F P D A G
6
7563


254
L S F P D A G F T
6
7564


281
A S P I F T D T V
6
7565


317
T C F V D A V A E
6
7566


322
A V A E L A R K A
6
7567


323
V A E L A R K A G
6
7568


327
A R K A G C K L T
6
7569


362
K T L P V V F D S
6
7570


382
K R I L G P D F G
6
7571


405
D S F G N L E V S
6
7572


436
G S S C R R V T Q
6
7573


473
E F L S F V P A P
6
7574


495
A C F K L F Q E K
6
7575


511
A L L F Q G V V D
6
7576


526
I S I N Q V L S N
6
7577


533
S N K D L I N Y N
6
7578


536
D L I N Y N K F V
6
7579


546
S C I D W N R E V
6
7580


38
E M F E V Y G T P
5
7581


41
E V Y G T P G V D
5
7582


51
Y I S P N N E R G
5
7583


61
E R A D T R R W R
5
7584


80
M N S P S N D L N
5
7585


82
S P S N D L N D S
5
7586


85
N D L N D S H V Q
5
7587


106
A Y A V L Y L T C
5
7588


128
R Q D R N F V D K
5
7589


145
G Y G G I L L V N
5
7590


156
R D D P S C D V Q
5
7591


178
E D M S V M V L R
5
7592


187
T Q G P A A L F D
5
7593


204
S S Y D A K R A Q
5
7594


224
E A Y R H V L G Q
5
7595


225
A Y R H V L G Q D
5
7596


228
H V L G Q D K V S
5
7597


247
E R F F V F C L S
5
7598


265
I S F H V T L L D
5
7599


282
S P I F T D T V V
5
7600


297
I M T P S T L P P
5
7601


302
T L P P L F V Y V
5
7602


330
A G C K L T I C P
5
7603


342
N R N D R W I Q D
5
7604


343
R N D R W I Q D E
5
7605


351
E M E L G Y V Q A
5
7606


359
A P H K T L P V V
5
7607


366
V V F D S P R N G
5
7608


375
E L Q D F P Y K R
5
7609


379
F P Y K R I L G P
5
7610


393
T R E P R D R S V
5
7611


395
E P R D R S V S G
5
7612


412
V S P P V V A N G
5
7613


428
I L I G G N L P G
5
7614


437
S S G R R V T Q V
5
7615


445
V V R D F L H A Q
5
7616


447
R D F L H A Q K V
5
7617


452
A Q K V Q P P V E
5
7618


455
V Q P P V E L F V
5
7619


476
S F V P A P D G K
5
7620


480
A P D G K G F R M
5
7621


490
L A S P G A C F K
5
7622


521
E Q V K T I S I N
5
7623


523
V K T I S I N Q V
5
7624


563
E C D I I D I P Q
5
7625


568
D I P Q L F K T E
5
7626


572
L F K T E R K K A
5
7627


573
F K T E R K K A T
5
7628


580
A T A F F P D L V
5
7629


594
G K H L G I P K P
5
7630


615
E K V R S L L E P
5
7631


643
E V H C G T N V C
5
7632


10
S L E H P T S A V
4
7633


14
P T S A V C V A G
4
7634


17
A V C V A G V E T
4
7635


27
V D I Y G S V P E
4
7636


47
G V D I Y I S P N
4
7637


48
V D I Y I S P N N
4
7638


57
E R G R E R A D T
4
7639


65
T R R W R F D A T
4
7640


81
N S P S N D L N D
4
7641


84
S N D L N D S H V
4
7642


112
L T C V D I S L D
4
7643


118
S L D C D L N C E
4
7644


126
E G R Q D R N F V
4
7645


181
S V M V L R T Q G
4
7646


194
F D D H K L V L H
4
7647


196
D H K L V L H T S
4
7648


203
T S S Y D A K R A
4
7649


205
S Y D A K R A Q V
4
7650


214
F H I C G P E D V
4
7651


216
I C G P E D V C E
4
7652


238
E V P R L H G D E
4
7653


255
S F P D A G F T G
4
7654


260
G F T G L I S F H
4
7655


262
T G L I S F H V T
4
7656


279
F S A S P I F T D
4
7657


287
D T V V F R V A P
4
7658


294
A P W I M T P S T
4
7659


299
T P S T L P P L E
4
7660


303
L P P L E V Y V C
4
7661


307
E V Y V C R V R N
4
7662


315
N N T C F V D A V
4
7663


325
E L A R K A G C K
4
7664


331
G C K L T I C P Q
4
7665


332
C K L T I C P Q A
4
7666


333
K L T I C P Q A E
4
7667


335
T I C P Q A E N R
4
7668


341
E N R N D R W I Q
4
7669


357
V Q A P H K T L P
4
7670


369
D S P R N G E L Q
4
7671


370
S P R N G E L Q D
4
7672


378
D F P Y K R I L G
4
7673


384
I L G P D F G Y V
4
7674


387
P D F G Y V T R E
4
7675


413
S P P V V A N G K
4
7676


418
A N G K E Y P L G
4
7677


424
P L G R I L I G G
4
7678


432
G N L P G S S G R
4
7679


435
P G S S G R R V T
4
7680


438
S G R R V T Q V V
4
7681


448
D F L H A Q K V Q
4
7682


488
M L L A S P G A C
4
7683


497
F K L F Q E K Q K
4
7684


498
K L F Q E K Q K C
4
7685


512
L L F Q G V V D D
4
7686


517
V V D D E Q V K T
4
7687


525
T I S I N Q V L S
4
7688


534
N K D L I N Y N K
4
7689


538
I N Y N K F V Q S
4
7690


548
I D W N R E V L K
4
7691


549
D W N R E V L K R
4
7692


571
Q L F K T E R K K
4
7693


581
T A F F P D L V N
4
7694


586
D L V N M L V L G
4
7695


588
V N M L V L G K H
4
7696


604
G P I I N G C C C
4
7697


610
C C C L E E K V R
4
7698


619
S L L E P L G L H
4
7699


620
L L E P L G L H C
4
7700


633
D F T P Y H M L H
4
7701


8
R V S L E H P T S
3
7702


16
S A V C V A G V E
3
7703


20
V A G V E T L V D
3
7704


31
G S V P E G T E M
3
7705


45
T P G V D I Y I S
3
7706


46
P G V D I Y I S P
3
7707


50
I Y I S P N M E R
3
7708


52
I S P N M E R G R
3
7709


54
P N M E R G R E R
3
7710


55
N M E R G R E R A
3
7711


58
R G R E R A D T R
3
7712


73
T L E I I V V M N
3
7713


78
V V M N S P S N D
3
7714


89
D S H V Q I S Y H
3
7715


92
V Q I S Y H S S H
3
7716


98
S S H E P L P L A
3
7717


110
L Y L T C V D I S
3
7718


113
T C V D I S L D C
3
7719


116
D I S L D C D L N
3
7720


117
I S L D C D L N C
3
7721


120
D C D L N C E G R
3
7722


124
N C E G R Q D R N
3
7723


127
G R Q D R N F V D
3
7724


129
Q D R N F V D K R
3
7725


132
N F V D K R Q W V
3
7726


140
V W G P S G Y G G
3
7727


153
N C D R D D P S C
3
7728


157
D D P S C D V Q D
3
7729


160
S C D V Q D N C D
3
7730


162
D V Q D N C D Q H
3
7731


179
D M S V M V L R T
3
7732


180
M S V M V L R T Q
3
7733


182
V M V L R T Q G P
3
7734


184
V L R T Q G P A A
3
7735


209
K R A Q V F H I C
3
7736


212
Q V F H I C C P E
3
7737


220
E D V C E A Y R H
3
7738


235
V S Y E V P R L H
3
7739


236
S Y S V P R L H G
3
7740


244
G D E E S F F V E
3
7741


266
S F H V T L L D D
3
7742


270
T L L D D S N E D
3
7743


27
D S N E D F S A S
3
7744


278
D F S A S P I F T
3
7745


284
I F T D T V V F R
3
7746


285
F T D T V V F R V
3
7747


286
T D T V V F R V A
3
7748


296
W I N T P S T L P
3
7749


300
P S T L P P L E V
3
7750


308
V Y V C R V R N N
3
7751


309
Y V C R V R N N T
3
7752


310
V C R V R N N T C
3
7753


312
R V R N N T C F V
3
7754


314
R N N T C F V D A
3
7755


319
F V D A V A E L A
3
7756


320
V D A V A E L A R
3
7757


33
L T I C P Q A E N
3
7758


338
P Q A E N R N D R
3
7759


355
G Y V Q A P H K T
3
7760


358
Q A P H K T L P V
3
7761


361
H K T L P V V F D
3
7762


367
V F D S P R N G E
3
7763


380
P Y K R I L G P D
3
7764


385
L G P D F G Y V T
3
7765


38
G P D F C Y V T R
3
7766


391
Y V T R E P R D R
3
7767


400
S V S G L D S F G
3
7768


403
G L D S F G N L E
3
7769


406
S F G N L E V S P
3
7770


409
N L E V S P P V V
3
7771


419
N G K E Y P L G R
3
7772


425
L G R I L I G G N
3
7773


427
R I L I G G N L P
3
7774


429
L I G G N L P G S
3
7775


433
N L P G S S G R R
3
7776


440
R R V T Q V V R D
3
7777


444
Q V V R D F L H A
3
7778


456
Q P P V F L F V D
3
7779


461
L F V D W L A V G
3
7780


462
F V D W L A V G H
3
7781


464
D W L A V G H V D
3
7782


475
L S F V P A P D G
3
7783


483
G K C F R M L L A
3
7784


486
F R N L L A S P G
3
7785


502
E K Q K C G H G R
3
7786


507
G H G R A L L F Q
3
7787


510
R A L L F Q G V V
3
7788


522
Q V K T I S I N Q
3
7789


531
V L S N K D L I N
3
7790


541
N K F V Q S C I D
3
7791


543
F V Q S C I D W N
3
7792


550
W N R E V L K R E
3
7793


554
V L K R E L G L A
3
7794


555
L K R E L G L A E
3
7795


556
K R E L G L A E C
3
7796


574
K T E R K K A T A
3
7797


578
K K A T A F F P D
3
7798


58
F P D L V N M L V
3
7799


590
M L V L G K H L G
3
7800


600
P K P F G P I I N
3
7801


617
V R S L L E P L G
3
7802


628
C T F I D D F T P
3
7803


630
F I D D F T P Y H
3
7804


637
Y H M L H G E V H
3
7805


639
M L H G E V H C G
3
7806


645
H C G T N V C R K
3
7807


646
C G T N V C R K P
3
7808


653
K P F S F K W W N
3
7809


655
F S F K W W N M V
3
7810


5
R I V R V S L E H
2
7811


9
V S L E H P T S A
2
7812


13
H P T S A V C V A
2
7813


19
C V A G V E T L V
2
7814


25
T L V D I Y G S V
2
7815


26
L V D I Y G S V P
2
7816


28
D I Y G S V P E G
2
7817


30
Y G S V P E G T E
2
7818


49
D I Y I S P N M E
2
7819


53
S P N M E R C R E
2
7820


64
D T R R W R F D A
2
7821


67
R W R F D A T L E
2
7822


70
F D A T L E I I V
2
7823


87
L N D S H V Q I S
2
7824


90
S H V Q I S Y H S
2
7825


96
Y H S S H E P L P
2
7826


105
L A Y A V L Y L T
2
7827


107
Y A V L Y L T C V
2
7828


114
C V D I S L D C D
2
7829


121
C D L N C E C R Q
2
7830


122
D L N C E G R Q D
2
7831


130
D R N F V D K R Q
2
7832


134
V D K R Q W V W G
2
7833


146
Y G G I L L V N C
2
7834


148
C I L L V N C D R
2
7835


149
I L L V N C D R D
2
7836


150
L L V N C D R D D
2
7837


152
V N C D R D D P S
2
7838


15
C D R D D P S C D
2
7839


155
D R D D P S C D V
2
7840


158
D P S C D V Q D N
2
7841


163
V Q D N C D Q H V
2
7842


16
Q D N C D Q H V H
2
7843


165
D N C D Q H V H C
2
7844


168
D Q H V H C L Q D
2
7845


171
V H C L Q D L E D
2
7846


172
H C L Q D L E D M
2
7847


174
L Q D L E D M S V
2
7848


175
Q D L E D M S V M
2
7849


176
D L E D M S V M V
2
7850


183
M V L R T Q G P A
2
7851


188
Q G P A A L F D D
2
7852


195
D D H K L V L H T
2
7853


201
L H T S S Y D A K
2
7854


202
H T S S Y D A K R
2
7855


207
D A K R A Q V F H
2
7856


215
H I C G P E D V C
2
7857


217
C G P E D V C S A
2
7858


227
R H V L G Q D K V
2
7859


230
L G Q D K V S Y E
2
7860


243
H G D E E R F F V
2
7861


261
F T G L I S F H V
2
7862


267
F H V T L L D D S
2
7863


269
V T L L D D S N E
2
7864


272
L D D S N E D F S
2
7865


273
D D S N F D F S A
2
7866


275
S N F D F S A S P
2
7867


293
V A P W I N T P S
2
7868


304
P P L E V Y V C R
2
7869


313
V R N N T C F V D
2
7870


316
N T C F V D A V A
2
7871


329
K A G C K L T I C
2
7872


336
I C P Q A E N R N
2
7873


344
N D R W I Q D E M
2
7874


347
W I Q D F M E L G
2
7875


353
E L G Y V Q A P H
2
7876


363
T L P V V F D S P
2
7877


388
D F G Y V T R E P
2
7878


390
G Y V T R F P R D
2
7879


398
D R S V S G L D S
2
7880


404
L D S F C N L S V
2
7881


407
F G N L E V S P P
2
7882


430
I G G N L P G S S
2
7883


439
G R R V T Q V V R
2
7884


443
T Q V V R D F L H
2
7885


446
V R D F L H A Q K
2
7886


449
F L H A Q K V Q P
2
7887


463
V D W L A V G H V
2
7888


470
H V D E F L S F V
2
7889


474
F L S F V P A P D
2
7890


477
F V P A P D G K G
2
7891


479
P A P D G K G F R
2
7892


487
R M L L A S P G A
2
7893


494
G A C F K L F Q E
2
7894


496
C F K L F Q E K Q
2
7895


499
L F Q E K Q K C G
2
7896


508
H G R A L L F Q G
2
7897


509
G R A L L F Q G V
2
7898


513
L F Q G V V D D E
2
7899


516
G V V D D E Q V K
2
7900


527
S I N Q V L S N K
2
7901


528
I N Q V L S N K D
2
7902


537
L I N Y N K F V Q
2
7903


539
N Y N K F V Q S C
2
7904


544
V Q S C I D W N R
2
7905


566
I I D I P Q L F K
2
7906


570
P Q L F K T E R K
2
7907


587
L V N M L V L C K
2
7908


603
F G P I I N G C C
2
7909


606
I I N C C C C L E
2
7910


608
N G C C C L E E K
2
7911


611
C C L E F K V R S
2
7912


624
L G L H C T F I D
2
7913


625
G L H C T F I D D
2
7914


631
I D D F T P Y H M
2
7915


634
F T P Y H M L H G
2
7916


635
T P Y H M L H G E
2
7917


638
H M L H G F V H C
2
7918


644
V H C G T N V C R
2
7919


656
S F K W W N M V P
2
7920


6
I V R V S L E H P
1
7921


7
V R V S L E H P T
1
7922


15
T S A V C V A C V
1
7923


29
I Y G S V P E G T
1
7924


33
V P E G T E M F E
1
7925


36
G T E M F E V Y G
1
7926


59
G R E R A D T R R
1
7927


77
I V V M N S P S N
1
7928


83
P S N D L N D S H
1
7929


91
H V Q I S Y H S S
1
7930


93
Q I S Y H S S H E
1
7931


94
I S Y H S S H E P
1
7932


119
L D C D L N C E G
1
7933


123
L N C E G R Q D R
1
7934


135
D K R Q W V W G P
1
7935


136
K R Q W V W G P S
1
7936


139
W V W G P S G Y G
1
7937


151
L V N C D R D D P
1
7938


159
P S C D V Q D N C
1
7939


167
C D Q H V H C L Q
1
7940


170
H V H C L Q D L E
1
7941


173
C L Q D L E D N S
1
7942


189
G P A A L F D D H
1
7943


190
P A A L F D D H K
1
7944


197
H K L V L H T S S
1
7945


199
L V L H T S S Y D
1
7946


200
V L H T S S Y D A
1
7947


210
R A Q V F H I C G
1
7948


221
D V C E A Y R H V
1
7949


231
G Q D K V S Y E V
1
7950


232
Q D K V S Y E V P
1
7951


233
D K V S Y E V P R
1
7952


239
V P R L H G D E E
1
7953


240
P R L H G D E E R
1
7954


249
F F V E G L S F P
1
7955


250
F V E G L S F P D
1
7956


258
D A G F T G L I S
1
7957


268
H V T L L D D S N
1
7958


290
V F R V A P W I M
1
7959


291
F R V A P W I M T
1
7960


305
P L E V Y V C R V
1
7961


321
D A V A B L A R K
1
7962


337
C P Q A E N R N D
1
7963


349
Q D S N E L C Y V
1
7964


354
L G Y V Q A P H K
1
7965


389
F G Y V T R E P R
1
7966


392
V T R E P R D R S
1
7967


397
R D R S V S G L D
1
7968


401
V S C L D S F G N
1
7969


408
G N L E V S P P V
1
7970


414
P P V V A N G K E
1
7971


416
V V A N G K E Y P
1
7972


431
G G N L P G S S G
1
7973


43
L P G S S G R R V
1
7974


450
L H A Q K V Q P P
1
7975


465
W L A V G H V D E
1
7976


468
V G H V D E F L S
1
7977


471
V D E F L S F V P
1
7978


485
G F R M L L A S P
1
7979


493
P G A C F K L F Q
1
7980


503
K Q K C G H G R A
1
7981


515
Q G V V D D E Q V
1
7982


545
Q S C I D W N R E
1
7983


560
G L A E C D I I D
1
7984


569
I P Q L F K T E R
1
7985


577
R K K A T A F F P
1
7986


592
V L G K H L G I P
1
7987


593
L G K H L G I P K
1
7988


607
I N G C C C L E E
1
7989


609
G C C C L E E K V
1
7990


627
H C T F I D D F T
1
7991


641
H G E V H C G T N
1
7992


648
T N V C R K P F S
1
7993


650
V C R K P F S F K
1
7994


654
P F S F K W W N M
1
7995







184P1E2 v.2: HLA Peptide Scoring Results B4402 9-


mers SYFPEITHI










6
S T L A P L E V Y
14
7996


3
M T P S T L A P L
12
7997


2
I M T P S T L A P
7
7998


9
A P L E V Y V C R
5
7999


4
T P S T L A P L E
4
8000


7
T L A P L E V Y V
4
8001


8
L A P L E V Y V C
4
8002


1
W I M T P S T L A
3
8003


5
P S T L A P L E V
3
8004







184P1E2 v.3: HLA Peptide Scoring Results B4402 9-


mers SYFPEITHI










1
D E F L S F V P V
15
8005


7
V P V P D G K G F
15
8006


2
E F L S F V P V P
5
8007


5
S F V P V P D G K
5
8008


3
F L S F V P V P D
3
8009


4
L S F V P V P D G
3
8010


6
F V P V P D G K G
2
8011


8
P V P D G K G F R
2
8012


9
V P D G K G F R M
2
8013
















TABLE XXXIV







184P1E2 v.1: HLA Peptide Scoring Results B5101 9-


mers SYFPEITHI













SEQ.





ID


Pos
1 2 3 4 5 6 7 8 9
score
NO.













71
D A T L E I I V V
27
8014


359
A P H K T L P V V
24
8015


561
L A E C D I I D I
24
8016


107
Y A V L Y L T C V
23
8017


282
S P I F T D T V V
23
8018


434
L P G S S G R R V
23
8019


510
R A L L F Q G V V
23
8020


584
F P D L V N M L V
22
8021


599
I P K P F G P I I
22
8022


21
A G V E T L V D I
21
8023


101
E P L P L A Y A V
21
8024


144
S G Y G G I L L V
21
8025


326
L A R K A G C K L
21
8026


358
Q A P H K T L P V
21
8027


451
H A G K V Q P P V
21
8028


559
L G L A E S C I I
21
8029


105
L A Y A V L Y L T
20
8030


191
A A L F D D H K L
20
8031


256
F P D A G F T G L
20
8032


417
V A N G K E Y P L
19
8033


438
S G R R V T Q V V
19
8034


103
L P L A Y A V L Y
18
8035


142
G P S G Y G G I L
18
8036


303
L P P L E V Y V C
18
8037


379
F P Y K R I L G P
18
8038


518
V D D A Q V K T I
18
8039


579
K A T A F F P D L
18
8040


86
D L N D S H V Q I
17
8041


109
V L Y L T C V D I
17
8042


141
W G P S G Y G G I
17
8043


224
E A Y R H V L G Q
17
8044


304
P P L E V Y V C R
17
8045


321
D A V A E L A R K
17
8046


402
S G L D S F G N L
17
8047


423
Y P L G R I L I G
17
8048


482
D G K G F R M L L
17
8049


591
L V L G K H L G I
17
8050


635
T P Y H M L H G E
17
8051


16
S A V C V A G V E
16
8052


158
D P S C D V Q D N
16
8053


221
D V C E A Y R H V
16
8054


243
H G D E E R F F V
16
8055


258
D A G F T G L I S
16
8056


520
D E Q V K T I S I
16
8057


581
T A F F P D L V N
16
8058


13
H P T S A V C V A
15
8059


20
V A G V F T L V D
15
8060


44
G T P G V D I Y I
15
8061


126
E G R Q D R N F V
15
8062


176
D L E D M S V M V
15
8063


207
D A K R A Q V F H
15
8064


285
F T D T V V F R V
15
8065


289
V V F R V A P W I
15
8066


413
S P P V V A N G K
15
8067


456
Q P P V F L F V D
15
8068


466
L A V G H V D E F
15
8069


536
D L I N Y N K F V
15
8070


622
E P L G L H C T F
15
8071


1
M S L Q R I V R V
14
8072


177
L E D M S V M V L
14
8073


208
A K R A Q V F H I
14
8074


262
T G L I S F H V T
14
8075


328
R K A G C K L T I
14
8076


354
L G Y V Q A P H K
14
8077


376
L Q D F P Y K R I
14
8078


408
G N L E V S P P V
14
8079


409
N L E V S P P V V
14
8080


457
P P V F L F V D W
14
8081


515
Q G V V D D E Q V
14
8082


540
Y N K F V Q S C I
14
8083


601
K P F G P I I N G
14
8084


12
E H P T S A V C V
13
8085


33
V P F G T E M F E
13
8086


35
E G T F M F E V Y
13
8087


42
V Y G T P G V D I
13
8088


68
W R F D A T L E I
13
8089


69
R F D A T L E I I
13
8090


155
D R D D P S C D V
13
8091


193
L F D D H K L V L
13
8092


218
G P F D V C F A Y
13
8093


276
N E D F S A S P I
13
8094


293
V A P W I M T P S
13
8095


299
T P S T L P P L E
13
8096


329
K A G C K L T I C
13
8097


337
C P Q A E N R N D
13
8098


385
L G P D F G Y V T
13
8099


386
G P D F G Y V T R
13
8100


414
P P V V A N G K E
13
8101


455
V Q P P V E L F V
13
8102


463
V D W L A V G H V
13
8103


480
A P D G K G F R M
13
8104


490
L A S P G A C F K
13
8105


492
S P G A C F K L F
13
8106


569
I P Q L F K T E R
13
8107


598
G I P K P F C P I
13
8108


18
V C V A G V E T L
12
8109


19
C V A G V E T L V
12
8110


28
D I Y G S V P S G
12
8111


45
T P G V D I Y I S
12
8112


62
R A D T R R W R F
12
8113


82
S P S N D L N D S
12
8114


189
G P A A L F D D H
12
8115


222
V C E A Y R H V L
12
8116


230
L G Q D K V S Y E
12
8117


280
S A S P I F T D T
12
8118


294
A P W I M T P S T
12
8119


305
P L E V Y V C R V
12
8120


323
V A S L A R K A G
12
8121


339
Q A E N R N D R W
12
8122


356
Y V Q A P H K T L
12
8123


364
L P V V F D S P R
12
8124


389
F G Y V T R S P R
12
8125


395
E P R D R S V S G
12
8126


404
L D S F G N L S V
12
8127


419
N G K E Y P L C R
12
8128


420
G K E Y P L C R I
12
8129


422
E Y P L G R I L I
12
8130


447
R D F L H A Q K V
12
8131


464
D W L A V G H V D
12
8132


478
V P A P D G K C F
12
8133


479
P A P D G K G F R
12
8134


494
G A C F K L F Q E
12
8135


523
V K T I S I N Q V
12
8136


530
Q V L S N K D L I
12
8137


583
F F P D L V N N L
12
8138


585
P D L V N M L V L
12
8139


612
C L E E K V R S L
12
8140


632
D D F T P Y H M L
12
8141


655
F S F K W W N M V
12
8142


3
L Q R I V R V S L
11
8143


15
T S A V C V A G V
11
8144


49
D I Y I S P N M E
11
8145


53
S P N M E R G R E
11
8146


58
R G R E R A D T R
11
8147


70
F D A T L S I I V
11
8148


111
Y L T C V D I S L
11
8149


132
N F V D K R Q W V
11
8150


190
P A A L F D D H K
11
8151


192
A L F D D H K L V
11
8152


210
R A Q V F H I C G
11
8153


217
C G P E D V C S A
11
8154


231
G Q D K V S Y E V
11
8155


235
V S Y E V P R L H
11
8156


239
V P R L H G D E E
11
8157


257
P D A G F T G L I
11
8158


264
L I S F H V T L L
11
8159


281
A S P I F T D T V
11
8160


302
T L P P L E V Y V
11
8161


315
N N T C F V D A V
11
8162


340
A E N R N D R W I
11
8163


370
S P R N C E L Q D
11
8164


384
I L C P D F G Y V
11
8165


421
K E Y P L G R I L
11
8166


437
S S G R R V T Q V
11
8167


470
H V D E F L S F V
11
8168


546
S C I D W N R S V
11
8169


589
N M L V L C K H L
11
8170


613
L E E K V R S L L
11
8171


623
P L G L H C T F I
11
8172


646
C G T N V C R K P
11
8173


653
K P F S F K W W N
11
8174


25
T L V D I Y G S V
10
8175


30
Y G S V P E G T E
10
8176


41
E V Y G T P C V D
10
8177


43
Y G T P C V D I Y
10
8178


102
P L P L A Y A V L
10
8179


146
Y G C I L L V N C
10
8180


163
V Q D N C D Q H V
10
8181


174
L Q D L E D M S V
10
8182


205
S Y D A K R A Q V
10
8183


214
F H I C G P E D V
10
8184


227
R H V L G Q D K V
10
8185


259
A G F T G L I S F
10
8186


261
F T C L I S F H V
10
8187


295
P W I M T P S T L
10
8188


298
M T P S T L P P L
10
8189


300
P S T L P P L E V
10
8190


393
T R E P R D R S V
10
8191


405
D S F G N L E V S
10
8192


435
P G S S G R R V T
10
8193


448
D F L H A Q K V Q
10
8194


460
E L F V D W L A V
10
8195


48
K G F R M L L A S
10
8196


491
A S P G A C F K L
10
8197


524
K T I S I N Q V L
10
8198


529
N Q V L S N K D L
10
8199


553
E V L K R E L G L
10
8200


558
E L G L A E C D I
10
8201


597
L G I P K P F G P
10
8202


604
G P I I N G C C C
10
8203


609
G C C C L E E K V
10
8204


618
R S L L E P L G L
10
8205


624
L G L H C T F I D
10
8206


34
P E G T E M F E V
9
8207


40
F E V Y G T P G V
9
8208


66
R R W R F D A T L
9
8209


72
A T L E I I V V M
9
8210


84
S N D L N D S H V
9
8211


10
P L A Y A V L Y L
9
8212


147
G G I L L V N C D
9
8213


166
N C D Q H V H C L
9
8214


179
D M S V M V L R T
9
8215


185
L R T Q G P A A L
9
8216


188
Q G P A A L F D D
9
8217


195
D D H K L V L H T
9
8218


196
D H K L V L H T S
9
8219


234
K V S Y E V P R L
9
8220


252
E G L S F P D A G
9
8221


263
G L I S F H V T L
9
8222


318
C F V D A V A E L
9
8223


349
Q D F N E L G Y V
9
8224


350
D E M E L G Y V Q
9
8225


377
Q D F P Y K R I L
9
8226


396
P R D R S V S G L
9
8227


407
F G N L F V S P P
9
8228


425
L G R I L I G G N
9
8229


430
I G G N L P G S S
9
8230


442
V T Q V V R D F L
9
8231


481
P D C K C F R M L
9
8232


506
C G H G R A L L F
9
8233


509
G R A L L F Q G V
9
8234


538
I N Y N K F V Q S
9
8235


547
C I D W N R E V L
9
8236


549
D W N R E V L K R
9
8237


568
D I P Q L F K T E
9
8238


580
A T A F F P D L V
9
8239


593
L G K H L G I P K
9
8240


603
F G P I I N G C C
9
8241


642
G E V H C G T N V
9
8242


10
S L E H P T S A V
8
8243


46
P G V D I Y I S P
8
8244


79
V M N S P S N D L
8
8245


9
H S S H E P L P L
8
8246


130
D R N F V D K R Q
8
8247


246
E E R F F V E G L
8
8248


278
D F S A S P I F T
8
8249


286
T D T V V F R V A
8
8250


306
L E V Y V C R V R
8
8251


312
R V R N N T C F V
8
8252


330
A G C K L T I C P
8
8253


368
F D S P R N G E L
8
8254


453
Q K V Q P P V E L
8
8255


458
P V F L F V D W L
8
8256


467
A V G H V D E F L
8
8257


472
D E F L S F V P A
8
8258


504
Q K C G H G R A L
8
8259


508
H G R A L L F Q G
8
8260


551
N R E V L K R E L
8
8261


608
N G C C C L E E K
8
8262


636
P Y H M L H G E V
8
8263


641
H Q E V H C G T N
8
8264


9
V S L E H P T S A
7
8265


38
E D F E V Y G T P
7
8266


94
I S Y H S S H E P
7
8267


95
S Y H S S H E P L
7
8268


110
L Y L T C V D I S
7
8269


115
V D I S L D C D L
7
8270


143
P S G Y C G I L L
7
8271


165
D N C D Q H V H C
7
8272


169
Q H V H C L Q D L
7
8273


175
Q D L E D N S V M
7
8274


180
M S V M V L R T Q
7
8275


206
Y D A K R A Q V F
7
8276


228
H V L G Q D K V S
7
8277


274
D S N E D F S A S
7
8278


284
I F T D T V V F R
7
8279


352
M E L G Y V Q A P
7
8280


373
N G E L Q D F P Y
7
8281


378
D F P Y K R I L G
7
8282


387
P D F G Y V T R E
7
8283


388
D F G Y V T R E P
7
8284


410
L E V S P P V V A
7
8285


461
L F V D W L A V G
7
8286


468
V G H V D E F L S
7
8287


493
P G A C F K L F Q
7
8288


513
L F Q G V V D D E
7
8289


519
D D E Q V K T I S
7
8290


550
W N R E V L K R E
7
8291


564
C D I I D I P Q L
7
8292


586
D L V N M L V L G
7
8293


616
K V R S L L E P L
7
8294


2
S L Q R I V R V S
6
8295


11
L E H P T S A V C
6
8296


26
L V D I Y G S V P
6
8297


74
L E I I V V M N S
6
8298


85
N D L N D S H V Q
6
8299


117
I S L D C D L N C
6
8300


135
D K R Q W V W G P
6
8301


145
G Y G G I L L V N
6
8302


157
D D P S C D V Q D
6
8303


20
S S Y D A K R A Q
6
8304


233
D K V S Y E V P R
6
8305


244
G D E E R F F V E
6
8306


245
D E E R F F V E G
6
8307


265
I S F H V T L L D
6
8308


283
P I F T D T V V F
6
8309


301
S T L P P L H V Y
6
8310


307
E V Y V C R V R N
6
8311


316
N T C F V D A V A
6
8312


322
A V A E L A R K A
6
8313


346
R W I Q D E M E L
6
8314


348
I Q D E M F L G Y
6
8315


360
P H K T L P V V F
6
8316


362
K T L P V V F D S
6
8317


366
V V F D S P R N G
6
8318


398
D R S V S G L D S
6
8319


412
V S P P V V A N G
6
8320


426
G R I L I G G N L
6
8321


431
C G N L P G S S G
6
8322


488
M L L A S P G A C
6
8323


499
L F Q E K Q K C G
6
8324


505
K C C H C R A L L
6
8325


511
A L L F Q G V V D
6
8326


512
L L F Q G V V D D
6
8327


565
D I I D I P Q L F
6
8328


571
Q L F K T E R K K
6
8329


572
L F K T E R K K A
6
8330


588
V N M L V L G K H
6
8331


605
P I I N G C C C L
6
8332


611
C C L E E K V R S
6
8333


640
L H G E V H C G T
6
8334


6
I V R V S L E H P
5
8335


29
I Y G S V P E G T
5
8336


52
I S P N M E R G R
5
8337


89
D S H V Q I S Y H
5
8338


108
A V L Y L T C V D
5
8339


122
D L N C E G R Q D
5
8340


127
G R Q D R N F V D
5
8341


133
F V D K R Q W V W
5
8342


148
G I L L V N C D R
5
8343


149
I L L V N C D R D
5
8344


162
D V Q D N C D Q H
5
8345


168
D Q H V H C L Q D
5
8346


199
L V L H T S S Y D
5
8347


270
T L L D D S N E D
5
8348


271
L L D D S N E D F
5
8349


287
D T V V F R V A P
5
8350


309
Y V C R V R N N T
5
8351


310
V C R V R N N T C
5
8352


313
V R N N T C F V D
5
8353


319
F V D A V A E L A
5
8354


336
I C P Q A E N R N
5
8355


345
D R W I Q D F M E
5
8356


363
T L P V V F D S P
5
8357


369
D S P R N C E L Q
5
8358


428
T L I G G N L P G
5
8359


433
N L P G S S G R R
5
8360


439
G R R V T Q V V R
5
8361


440
R R V T Q V V R D
5
8362


441
R V T Q V V R D F
5
8363


450
L H A Q K V Q P P
5
8364


459
V E L F V D W L A
5
8365


477
F V P A P D G K G
5
8366


517
V V D D E Q V K T
5
8367


526
I S I N Q V L S N
5
8368


528
I N Q V L S N K D
5
8369


532
L S N K D L I N Y
5
8370


535
K D L I N Y N K F
5
8371


537
L I N Y N K F V Q
5
8372


555
L K R E L G L A E
5
8373


567
I D I P Q L F K T
5
8374


575
T E R K K A T A F
5
8375


587
L V N M L V L G K
5
8376


595
K H L G I P K P F
5
8377


619
S L L E P L G L H
5
8378


631
I D D F T P Y H M
5
8379


633
D F T P Y H M L H
5
8380


637
Y H M L H G E V H
5
8381


644
V H C G T N V C R
5
8382


1
P T S A V C V A G
4
8383


23
V E T L V D I Y G
4
8384


6
D T R R W R F D A
4
8385


73
T L E I I V V M N
4
8386


76
I I V V M N S P S
4
8387


87
L N D S H V Q I S
4
8388


88
N D S H V Q I S Y
4
8389


116
D I S L D C D L N
4
8390


119
L D C D L N C E G
4
8391


120
D C D L N C E G R
4
8392


123
L N C E G R Q D R
4
8393


129
Q D R N F V D K R
4
8394


156
R D D P S C D V Q
4
8395


183
M V L R T Q G P A
4
8396


194
F D D H K L V L H
4
8397


203
T S S Y D A K R A
4
8398


215
H I C G P E D V C
4
8399


232
Q D K V S Y E V P
4
8400


242
L H G D E E R F F
4
8401


249
F F V E G L S F P
4
8402


25
L S F P D A G F T
4
8403


255
S F P D A G F T G
4
8404


273
D D S N E D F S A
4
8405


279
F S A S P I F T D
4
8406


297
I M T P S T L P P
4
8407


308
V Y V C R V R N N
4
8408


317
T C F V D A V A E
4
8409


375
E L Q D F P Y K R
4
8410


391
Y V T R E P R D R
4
8411


392
V T S S P R D S S
4
8412


394
R E P R D S S V S
4
8413


429
L I G G N L P C S
4
8414


436
G S S C S R V T Q
4
8415


452
A Q K V Q P P V E
4
8416


471
V D E F L S F V P
4
8417


473
E F L S F V P A P
4
8418


474
F L S F V P A P D
4
8419


475
L S F V P A P D G
4
8420


497
F K L F Q E K Q K
4
8421


516
G V V D D E Q V K
4
8422


539
N Y N K F V Q S C
4
8423


548
I D W N R E V L K
4
8424


557
R E L G L A E C D
4
8425


570
P Q L F K T E R K
4
8426


573
F K T E R K K A T
4
8427


610
C C C L E E K V R
4
8428


620
L L E P L G L H C
4
8429


621
L E P L G L H C T
4
8430


626
L H C T F I D D F
4
8431


629
T F I D D F T P Y
4
8432


634
F T P Y H M L H G
4
8433


638
H M L H G E V H C
4
8434


639
M L H G E V H C G
4
8435


643
E V H C G T N V C
4
8436


645
H C G T N V C R K
4
8437


652
R K P F S F K W W
4
8438


656
S F K W W N M V P
4
8439


5
R I V R V S L E H
3
8440


22
G V E T L V D I Y
3
8441


24
E T L V D T Y G S
3
8442


32
S V P E G T E M F
3
8443


37
T E M F E V Y G T
3
8444


39
M F E V Y G T P G
3
8445


50
I Y I S P N M E R
3
8446


51
Y I S P N M E R G
3
8447


54
P N M E R G R E R
3
8448


56
M E R G R E R A D
3
8449


60
R E R A D T R R W
3
8450


61
E R A D T R R W R
3
8451


75
E I I V V M N S P
3
8452


78
V V M N S P S N D
3
8453


80
M N S P S N D L N
3
8454


81
N S P S N D L N D
3
8455


98
S S H S P L P L A
3
8456


106
A Y A V L Y L T C
3
8457


112
L T C V D I S L D
3
8458


113
T C V D I S L D C
3
8459


121
C D L N C E C R Q
3
8460


124
N C E G R Q D R N
3
8461


I50
L L V N C D R D D
3
8462


159
P S C D V Q D N C
3
8463


16
Q D N C D Q H V H
3
8464


172
H C L Q D L E D M
3
8465


178
E D M S V M V L R
3
8466


187
T Q G P A A L F D
3
8467


197
H K L V L H T S S
3
8468


200
V L H T S S Y D A
3
8469


201
L H T S S Y D A K
3
8470


202
H T S S Y D A K R
3
8471


209
K R A Q V F H I C
3
8472


216
I C G P E D V C E
3
8473


225
A Y R H V L G Q D
3
8474


237
Y E V P R L H G D
3
8475


247
E R F F V E G L S
3
8476


251
V E G L S F P D A
3
8477


266
S F H V T L L D D
3
8478


267
F H V T L L D D S
3
8479


269
V T L L D D S N E
3
8480


288
T V V F R V A P W
3
8481


291
F R V A P W I M T
3
8482


292
R V A P W I M T P
3
8483


314
R N N T C F V D A
3
8484


324
A E L A R K A G C
3
8485


332
C K L T I C P Q A
3
8486


338
P Q A E N R N D R
3
8487


342
N R N D R W I Q D
3
8488


355
G Y V Q A P H K T
3
8489


361
H K T L P V V F D
3
8490


374
G E L Q D F P Y K
3
8491


383
R I L G P D F G Y
3
8492


411
E V S P P V V A N
3
8493


427
R I L I G G N L P
3
8494


444
Q V V R D F L H A
3
8495


445
V V R D F L H A Q
3
8496


449
F L H A Q K V Q P
3
8497


462
F V D W L A V G H
3
8498


469
G H V D E F L S F
3
8499


495
A C F K L F Q E K
3
8500


498
K L F Q F K Q K C
3
8501


514
F Q G V V D D E Q
3
8502


521
F Q V K T I S I N
3
8503


527
S I N Q V L S N K
3
8504


531
V L S N K D L I N
3
8505


533
S N K D L I N Y N
3
8506


560
C L A E C D I I D
3
8507


566
I I D I P Q L F K
3
8508


574
K T E R K K A T A
3
8509


577
R K K A T A F F P
3
8510


582
A F F P D L V N M
3
8511


592
V L G K H L G I P
3
8512


594
G K H L G I P K P
3
8513


600
P K P F C P I I N
3
8514


606
I I N G C C C L E
3
8515


607
I N G C C C L E E
3
8516


615
E K V R S L L E P
3
8517


625
G L H C T F I D D
3
8518


628
C T F I D D F T P
3
8519


649
N V C R K P F S F
3
8520


650
V C R K P F S F K
3
8521


4
Q R I V R V S L E
2
8522


8
R V S L E H P T S
2
8523


27
V D I Y G S V P E
2
8524


47
C V D I Y I S P N
2
8525


55
N M E R G R E R A
2
8526


59
G R E R A D T R R
2
8527


65
T R R W R F D A T
2
8528


67
R W R F D A T L E
2
8529


77
I V V M N S P S N
2
8530


83
P S N D L N D S H
2
8531


92
V Q I S Y H S S H
2
8532


96
Y H S S H E P L P
2
8533


99
S H E P L P L A Y
2
8534


100
H E P L P L A Y A
2
8535


114
C V D I S L D C D
2
8536


118
S L D C D L N C E
2
8537


128
R Q D R N F V D K
2
8538


131
R N F V D K R Q W
2
8539


134
V D K R Q W V W G
2
8540


136
K R Q W V W G P S
2
8541


139
W V W G P S G Y G
2
8542


140
V W G P S G Y G G
2
8543


151
L V N C D R D D P
2
8544


152
V N C D R D D P S
2
8545


154
C D R D D P S C D
2
8546


167
C D Q H V H C L Q
2
8547


171
V H C L Q D L E D
2
8548


184
V L R T Q C P A A
2
8549


186
F T Q C P A A L F
2
8550


220
E D V C F A Y R H
2
8551


226
Y R H V L G Q D K
2
8552


229
V L G Q D K V S Y
2
8553


236
S Y E V P R L H G
2
8554


240
P R L H G D E E R
2
8555


241
R L H G D E E R F
2
8556


248
R F F V E G L S F
2
8557


250
F V E G L S F P D
2
8558


260
G F T G L I S F H
2
8559


272
L D D S N E D F S
2
8560


290
V F R V A P W I M
2
8561


296
W I M T P S T L P
2
8562


311
C R V R N N T C F
2
8563


320
V D A V A E L A R
2
8564


325
E L A R K A G C K
2
8565


327
A R K A G C K L T
2
8566


331
G C K L T I C P Q
2
8567


334
L T I C P Q A E N
2
8568


335
T I C P Q A E N R
2
8569


353
E L G Y V Q A P H
2
8570


357
V Q A P H K T L P
2
8571


367
V F D S P R N G E
2
8572


371
P R N C E L Q D F
2
8573


372
R N G E L Q D F P
2
8574


381
Y K R I L G P D F
2
8575


390
C Y V T R F P R D
2
8576


397
R D R S V S G L D
2
8577


415
P V V A N G K E Y
2
8578


424
P L G R I L I G G
2
8579


432
G N L P G S S G R
2
8580


454
K V Q P P V E L F
2
8581


486
F R M L L A S P G
2
8582


487
R M L L A S P G A
2
8583


489
L L A S P G A C F
2
8584


496
C F K L F Q E K Q
2
8585


500
F Q E K Q K C C H
2
8586


501
Q E K Q K C C H G
2
8587


507
G H G R A L L F Q
2
8588


522
Q V K T I S I N Q
2
8589


525
T I S I N Q V L S
2
8590


534
N K D L I N Y N K
2
8591


541
N K F V Q S C I D
2
8592


542
K F V Q S C I D W
2
8593


543
F V Q S C I D W N
2
8594


545
Q S C I D W N R E
2
8595


554
V L K R E L C L A
2
8596


562
A E C D I I D I P
2
8597


578
K K A T A F F P D
2
8598


602
P F C P I I N C C
2
8599


630
F I D D F T P Y H
2
8600


651
C R K P F S F K W
2
8601


7
V R V S L F H P T
1
8602


17
A V C V A G V E T
1
8603


31
G S V P E G T E M
1
8604


3
G T E M F E V Y G
1
8605


48
V D T Y I S P N M
1
8606


63
A D T R R W R F D
1
8607


90
S H V Q I S Y H S
1
8608


91
H V Q I S Y H S S
1
8609


93
Q I S Y H S S H E
1
8610


125
C E G R Q D R N F
1
8611


137
R Q W V W G P S G
1
8612


138
Q W V W G P S G Y
1
8613


153
N C D R D D P S C
1
8614


160
S C D V Q D N C D
1
8615


161
C D V Q D N C D Q
1
8616


173
C L Q D L E D M S
1
8617


181
S V M V L R T Q G
1
8618


182
V M V L R T Q C P
1
8619


211
A Q V F H I C G P
1
8620


212
Q V F H I C G P E
1
8621


213
V F H I C C P E D
1
8622


223
C E A Y R H V L G
1
8623


238
E V P R L H G D E
1
8624


275
S N E D F S A S P
1
8625


277
E D F S A S P I F
1
8626


333
K L T I C P Q A E
1
8627


343
R N D R W I Q D E
1
8628


344
N D R W I Q D E M
1
8629


347
W I Q D E M E L G
1
8630


351
E M E L G Y V Q A
1
8631


365
P V V F D S P R N
1
8632


380
P Y K R I L C P D
1
8633


382
K R I L G P D F G
1
8634


399
R S V S G L D S F
1
8635


400
S V S G L D S F G
1
8636


401
V S C L D S F G N
1
8637


403
G L D S F G N L E
1
8638


406
S F G N L E V S P
1
8639


416
V V A N C K E Y P
1
8640


418
A N G K E Y P L G
1
8641


443
T Q V V R D F L H
1
8642


446
V R D F L H A Q K
1
8643


465
W L A V G H V D E
1
8644


476
S F V P A P D G K
1
8645


483
G K C F R M L L A
1
8646


485
C F R M L L A S P
1
8647


502
E K Q K C G H G R
1
8648



















TABLE XXXV








SEQ.





ID


Pos
1 2 3 4 5 6 7 8 9 0
score
NO.















184P1E2 v.1: HLA Peptide Scoring Results A1 10-mers


SYFPEITHI










87
L N D S H V Q I S Y
29
8649


98
S S H E P L P L A Y
27
8650


628
C T F I D D F T P Y
24
8651


285
F T D T V V F R V A
21
8652


347
W I Q D E M E L G Y
21
8653


531
V L S N K D L I N Y
21
8654


300
P S T L P P L E V Y
20
8655


34
P E G T E M F E V Y
19
8656


102
P L P L A Y A V L Y
18
8657


194
F D D H K L V L H T
18
8658


403
G L D S F G N L E V
18
8659


584
F P D L V N M L V L
18
8660


613
L E E K V R S L L E
18
8661


21
A G V E T L V D I Y
17
8662


36
G T E M F E V Y G T
17
8663


42
V Y G T P G V D I Y
17
8664


319
F V D A V A E L A R
17
8665


99
S H E P L P L A Y A
16
8666


228
H V L G Q D K V S Y
16
8667


393
T R E P R D R S V S
16
8668


414
P P V V A N G K E Y
16
8669


480
A P D G K G F R M L
16
8670


574
K T E R K K A T A F
16
8671


137
R Q W V W G P S G Y
15
8672


197
H K L V L H T S S Y
15
8673


217
C G P E D V C E A Y
15
8674


250
F V E G L S F P D A
15
8675


372
R N G E L Q D F P Y
15
8676


382
K R I L G P D F G Y
15
8677


620
L L E P L G L H C T
15
8678


10
S L E H P T S A V C
14
8679


22
G V E T L V D I Y G
14
8680


69
R F D A T L E I I V
14
8681


118
S L D C D L N C E G
14
8682


156
R D D P S C D V Q D
14
8683


243
H G D E E R F F V E
14
8684


471
V D E F L S F V P A
14
8685


519
D D E Q V K T I S I
14
8686


580
A T A F F P D L V N
14
8687


612
C L E E K V R S L L
14
8688


84
S N D L N D S H V Q
13
8689


128
R Q D R N F V D K R
13
8690


160
S C D V Q D N C D Q
13
8691


176
D L E D M S V M V L
13
8692


186
R T Q G P A A L F D
13
8693


205
S Y D A K R A Q V F
13
8694


222
V C E A Y R H V L G
13
8695


244
G D E E R F F V E G
13
8696


256
F P D A G F T G L I
13
8697


362
K T L P V V F D S P
13
8698


373
N G E L Q D F P Y K
13
8699


446
V R D F L H A Q K V
13
8700


551
N R E V L K R E L G
13
8701


33
V P E G T E M F E V
12
8702


73
T L E I I V V M N S
12
8703


112
L T C V D I S L D C
12
8704


143
P S G Y G G I L L V
12
8705


163
V Q D N C D Q H V H
12
8706


166
N C D Q H V H C L Q
12
8707


177
L E D M S V M V L R
12
8708


193
L F D D H K L V L H
12
8709


235
V S Y F V P R L H C
12
8710


236
S Y E V P R L H G D
12
8711


275
S N E D F S A S P I
12
8712


301
S T L P P L E V Y V
12
8713


305
P L E V Y V C R V R
12
8714


349
Q D E M E L G Y V Q
12
8715


367
V F D S P R N G E L
12
8716


396
P R D R S V S G L D
12
8717


409
N L E V S P P V V A
12
8718


458
P V E L F V D W L A
12
8719


462
F V D W L A V G H V
12
8720


517
V V D D F Q V K T I
12
8721


534
N K D L I N Y N K F
12
8722


547
C I D W N R F V L K
12
8723


566
I I D I P Q L F K T
12
8724


124
N C E G R Q D R N F
11
8725


133
F V D K R Q W V W C
11
8726


155
D R D D P S C D V Q
11
8727


174
L Q D L E D N S V M
11
8728


231
G Q D K V S Y E V P
11
8729


245
D E E R F F V E G L
11
8730


261
F T G L I S F H V T
11
8731


271
L L D D S N E D F S
11
8732


323
V A E L A R K A G C
11
8733


351
E M E L G Y V Q A P
11
8734


357
V Q A P H K T L P V
11
8735


377
Q D F P Y K R I L G
11
8736


386
G P D F G Y V T R E
11
8737


420
G K E Y P L G R I L
11
8738


421
K E Y P L G R I L I
11
8739


470
H V D E F L S F V P
11
8740


491
A S P G A C F K L F
11
8741


500
F Q E K Q K C G H G
11
8742


518
V D D S Q V K T I S
11
8743


630
F I D D F T P Y H M
11
8744


631
I D D F T P Y H M L
11
8745


641
H G E V H C G T N V
11
8746


26
L V D I Y G S V P E
10
8747


39
M F E V Y G T P G V
10
8748


43
Y G T P G V D I Y I
10
8749


47
G V D I Y I S P N M
10
8750


55
N M E K G K S K A D
10
8751


59
G R E R A D T R R W
10
8752


62
R A D T R R W R F D
10
8753


80
M N S P S N D L N D
10
8754


114
C V D I S L D C D L
10
8755


120
D C D L N C E G R Q
10
8756


144
S C Y G G I L L V N
10
8757


153
N C D R D D P S C D
10
8758


218
G P E D V C E A Y R
10
8759


219
P S D V C E A Y R H
10
8760


265
I S F H V T L L D D
10
8761


272
L D D S N E D F S A
10
8762


276
N E D F S A S P I F
10
8763


339
Q A E N R N D R W I
10
8764


343
K N D K W I Q D E M
10
8765


348
I Q D E M E L G Y V
10
8766


369
D S P R N C E L Q D
10
8767


376
L Q D F P Y K K I L
10
8768


454
K V Q P P V S L F V
10
8769


468
V G H V D E F L S F
10
8770


556
K R E L G L A F C D
10
8771


561
L A E C D I I D I P
10
8772


563
E C D I I D I P Q L
10
8773


619
S L L E P L G L H C
10
8774


633
D F T P Y H M L H G
10
8775


31
G S V P E G T E M F
9
8776


64
D T R K W K F D A T
9
8777


72
A T L E I I V V M N
9
8778


192
A L F D D H K L V L
9
8779


254
L S F P D A C F T C
9
8780


412
V S P P V V A N G K
9
8781


442
V T Q V V R D F L H
9
8782


443
T Q V V R D F L H A
9
8783


482
D G K G F R M L L A
9
8784


548
I D W N R S V L K R
9
8785


55
V L K R E L G L A E
9
8786


19
C V A G V S T L V D
8
8787


96
Y H S S H E P L P L
8
8788


142
G P S G Y G G I L L
8
8789


257
P D A G F T G L I S
8
8790


264
L I S F H V T L L D
8
8791


279
F S A S P I F T D T
8
8792


298
M T P S T L P P L E
8
8793


368
F D S P R N G E L Q
8
8794


401
V S G L D S F G N L
8
8795


476
S F V P A P D G K G
8
8796


492
S P G A C F K L F Q
8
8797


505
K C G H G R A L L F
8
8798


524
K T I S I N Q V L S
8
8799


586
D L V N M L V L G K
8
8800


599
I P K P F G P I I N
8
8801


14
P T S A V C V A G V
7
8802


44
G T P G V D I Y I S
7
8803


103
L P L A Y A V L Y L
7
8804


178
E D M S V M V L R T
7
8805


204
S S Y D A K K A Q V
7
8806


223
C S A Y K H V L G Q
7
8807


247
E R F F V S G L S F
7
8808


269
V T L L D D S N E D
7
8809


287
D T V V F R V A P W
7
8810


302
T L P P L E V Y V C
7
8811


327
A K K A G C K L T I
7
8812


384
I L G P D F C Y V T
7
8813


392
V T R E P R D R S V
7
8814


397
R D R S V S C L D S
7
8815


418
A N G K S Y P L G R
7
8816


423
Y P L C K I L I G G
7
8817


427
R I L I G C N L P G
7
8818


437
S S G R R V T Q V V
7
8819


459
V E L F V D W L A V
7
8820


506
C C H G R A L L F Q
7
8821


525
T I S I N Q V L S N
7
8822


567
I D I P Q L F K T S
7
8823


582
A F F P D L V N M L
7
8824


590
M L V L G K H L G I
7
8825


592
V L G K H L C I P K
7
8826


597
L G I P K P F G P I
7
8827


606
I I N G C C C L E E
7
8828


617
V R S L L E P L G L
7
8829


62
L G L H C T F I D D
7
8830


634
F T P Y H M L H G K
7
8831


651
C R K P F S F K W W
7
8832


655
F S F K W W N M V P
7
8833


1
M S L Q R I V R V S
6
8834


4
Q R I V R V S L E H
6
8835


24
E T L V D I Y G S V
6
8836


51
Y I S P N M K R G R
6
8837


67
R W R F D A T L E I
6
8838


70
F D A T L E I I V V
6
8839


97
H S S H K P L P L A
6
8840


105
L A Y A V L Y L T C
6
8841


116
D I S L D C D L N C
6
8842


167
C D Q H V H C L Q D
6
8843


170
H V H C L Q D L E D
6
8844


202
H T S S Y D A K R A
6
8845


237
Y E V P R L H G D E
6
8846


280
S A S P I F T D T V
6
8847


281
A S P I F T D T V V
6
8848


290
V F R V A P W I H T
6
8849


296
W I M T P S T L P P
6
8850


299
T P S T L P P L E V
6
8851


316
N T C F V D A V A E
6
8852


334
L T I C P Q A E N R
6
8853


378
D F P Y K R I L G P
6
8854


422
E Y P L C K I L I C
6
8855


436
C S S G R R V T Q V
6
8856


455
V Q P P V E L F V D
6
8857


483
C K G F R M L L A S
6
8858


490
L A S P G A C F K L
6
8859


530
Q V L S N K D L I N
6
8860


552
K E V L K R K L G L
6
8861


565
D I I D I P Q L F K
6
8862


614
E E K V R S L L E P
6
8863


618
R S L L K P L C L H
6
8864


647
C T N V C R K P F S
6
8865


9
V S L E H P T S A V
5
8866


15
T S A V C V A C V E
5
8867


52
I S P N M E K C R E
5
8868


101
E P L P L A Y A V L
5
8869


111
Y L T C V D I S L D
5
8870


140
V W G P S C Y C G I
5
8871


180
N S V M V L R T Q C
5
8872


216
I C G P E D V C E A
5
8873


255
S F P D A C F T G L
5
8874


402
S C L D S F G N L K
5
8875


405
D S F G N L E V S P
5
8876


411
E V S P P V V A N C
5
8877


413
S P P V V A N G K E
5
8878


478
V P A P D G K G F R
5
8879


545
Q S C I D W N R E V
5
8880


560
G L A K C D I I D I
5
8881


562
A E C D I I D I P Q
5
8882


602
P F G P I I N G C C
5
8883


2
S L Q R I V R V S L
4
8884


11
L E H P T S A V C V
4
8885


45
T P G V D I Y I S P
4
8886


79
V N N S P S N D L N
4
8887


81
N S P S N D L N D S
4
8888


83
P S N D L N D S H V
4
8889


89
D S H V Q I S Y H S
4
8890


94
I S Y H S S H E P L
4
8891


95
S Y H S S H E P L P
4
8892


110
L Y L T C V D I S L
4
8893


117
I S L D C D L N C K
4
8894


159
P S C D V Q D N C D
4
8895


187
T Q G P A A L F D D
4
8896


203
T S S Y D A K R A Q
4
8897


208
A K R A Q V F H I C
4
8898


209
K R A Q V F H I C G
4
8899


226
Y R H V L G Q D K V
4
8900


258
D A G F T G L I S F
4
8901


263
G L I S F H V T L L
4
8902


274
D S N E D F S A S P
4
8903


277
K D F S A S P I F T
4
8904


292
R V A P W I M T P S
4
8905


297
I N T P S T L P P L
4
8906


313
V R N N T C F V D A
4
8907


325
E L A R K A G C K L
4
8908


329
K A G C K L T I C P
4
8909


335
T I C P Q A K N R N
4
8910


341
K N R N D K W I Q D
4
8911


356
Y V Q A P H K T L P
4
8912


383
K I L C P D F C Y V
4
8913


399
K S V S C L D S F C
4
8914


417
V A N C K E Y P L C
4
8915


432
C N L P C S S C R K
4
8916


475
L S F V P A P D G K
4
8917


504
Q K C C H C K A L L
4
8918


521
E Q V K T I S I N Q
4
8919


526
I S I N Q V L S N K
4
8920


527
S I N Q V L S N K D
4
8921


532
L S N K D L I N Y N
4
8922


541
N K F V Q S C I D W
4
8923


546
S C I D W N R E V L
4
8924


559
L G L A F C D I I D
4
8925


571
Q L F K T K R K K A
4
8926


583
F F P D L V N M L V
4
8927


600
P K P F G P I I N G
4
8928


3
L Q R I V R V S L E
3
8929


16
S A V C V A C V E T
3
8930


18
V C V A C V S T L V
3
8931


25
T L V D I Y C S V P
3
8932


30
Y C S V P E G T E M
3
8933


53
S P N M E R G R E R
3
8934


82
S P S N D L N D S H
3
8935


104
P L A Y A V L Y L T
3
8936


107
Y A V L Y L T C V D
3
8937


109
V L Y L T C V D I S
3
8938


115
V D I S L D C D L N
3
8939


127
C R Q D R N F V D K
3
8940


146
Y C G I L L V N C D
3
8941


188
Q G P A A L F D D H
3
8942


191
A A L F D D H K L V
3
8943


229
V L G Q D K V S Y E
3
8944


273
D D S N F D F S A S
3
8945


282
S P I F T D T V V F
3
8946


309
Y V C K V R N N T C
3
8947


391
Y V T R F P K D R S
3
8948


400
S V S G L D S F G N
3
8949


406
S F G N L E V S P P
3
8950


426
C R I L I C G N L P
3
8951


433
N L P C S S C R R V
3
8952


438
S C R R V T Q V V K
3
8953


449
F L H A Q K V Q P P
3
8954


453
Q K V Q P P V F L F
3
8955


456
Q P P V F L F V D W
3
8956


463
V D W L A V G H V D
3
8957


467
A V G H V D E F L S
3
8958


481
P D G K C F R M L L
3
8959


494
C A C F K L F Q E K
3
8960


495
A C F K L F Q F K Q
3
8961


497
F K L F Q E K Q K C
3
8962


508
H G R A L L F Q G V
3
8963


511
A L L F Q G V V D D
3
8964


523
V K T I S I N Q V L
3
8965


533
S N K D L I N Y N K
3
8966


537
L I N Y N K F V Q S
3
8967


577
R K K A T A F F P D
3
8968


578
K K A T A F F P D L
3
8969


644
V H C G T N V C R K
3
8970


649
N V C R K P F S F K
3
8971


650
V C R K P F S F K W
3
8972


12
E H P T S A V C V A
2
8973


17
A V C V A C V E T L
2
8974


27
V D I Y G S V P E G
2
8975


32
S V P S G T S M F S
2
8976


40
F E V Y G T P C V D
2
8977


41
E V Y G T P C V D I
2
8978


48
V D I Y I S P N M S
2
8979


56
M E R G R E R A D T
2
8980


63
A D T R R W R F D A
2
8981


66
R R W R F D A T L E
2
8982


86
D L N D S H V Q I S
2
8983


90
S H V Q I S Y H S S
2
8984


122
D L N C F C R Q D R
2
8985


126
E G R Q D R N F V D
2
8986


134
V D K R Q W V W G P
2
8987


139
W V W G P S G Y G C
2
8988


150
L L V N C D K D D P
2
8989


169
Q H V H C L Q D L F
2
8990


181
S V M V L R T Q G P
2
8991


184
V L R T Q G P A A L
2
8992


185
L K T Q C P A A L F
2
8993


190
P A A L F D D H K L
2
8994


200
V L H T S S Y D A K
2
8995


201
L H T S S Y D A K R
2
8996


206
Y D A K R A Q V F H
2
8997


214
F H I C G P E D V C
2
8998


221
D V C E A Y R H V L
2
8999


225
A Y R H V L G Q D K
2
9000


234
K V S Y F V P R L H
2
9001


241
R L H G D E F R F F
2
9002


246
E E R F F V F C L S
2
9003


266
S F H V T L L D D S
2
9004


270
T L L D D S N E D F
2
9005


295
P W I M T P S T L P
2
9006


308
V Y V C R V K N N T
2
9007


310
V C R V R N N T C F
2
9008


318
C F V D A V A E L A
2
9009


321
D A V A E L A R K A
2
9010


322
A V A E L A R K A G
2
9011


326
L A R K A C C K L T
2
9012


340
A E N R N D R W I Q
2
9013


346
R W I Q D E M K L C
2
9014


354
L G Y V Q A P H K T
2
9015


359
A P H K T L P V V F
2
9016


366
V V F D S P R N G K
2
9017


370
S P R N G K L Q D F
2
9018


374
C K L Q D F P Y K R
2
9019


381
Y K R I L G P D F G
2
9020


428
I L I G C N L P G S
2
9021


434
L P G S S C R R V T
2
9022


460
E L F V D W L A V G
2
9023


465
W L A V G H V D E F
2
9024


474
F L S F V P A P D G
2
9025


477
F V P A P D G K G F
2
9026


488
M L L A S P G A C F
2
9027


510
R A L L F Q G V V D
2
9028


512
L L F Q G V V D D K
2
9029


516
C V V D D E Q V K T
2
9030


529
N Q V L S N K D L I
2
9031


536
D L I N Y N K F V Q
2
9032


540
Y N K F V Q S C I D
2
9033


553
E V L K R K L C L A
2
9034


564
C D I I D I P Q L F
2
9035


570
P Q L F K T E R K K
2
9036


579
K A T A F F P D L V
2
9037


585
P D L V N M L V L G
2
9038


587
L V N M L V L G K H
2
9039


588
V N M L V L C K H L
2
9040


589
N M L V L G K H L C
2
9041


593
L C K H L G I P K P
2
9042


603
F G P I I N G C C C
2
9043


605
P I I N G C C C L K
2
9044


608
N C C C C L E E K V
2
9045


616
K V R S L L K P L G
2
9046


632
D D F T P Y H M L H
2
9047


636
P Y H M L H G E V H
2
9048


637
Y H M L H G E V H C
2
9049


639
M L H C K V H C G T
2
9050


645
H C G T N V C R K P
2
9051


7
V R V S L E H P T S
1
9052


20
V A G V E T L V D I
1
9053


23
V K T L V D I Y G S
1
9054


28
D I Y C S V P E G T
1
9055


38
E M F E V Y C T P C
1
9056


49
D I Y I S P N M E R
1
9057


54
P N H E R C R E R A
1
9058


60
R E R A D T R R W R
1
9059


61
E R A D T R R W R F
1
9060


71
D A T L E I I V V N
1
9061


75
K I I V V M N S P S
1
9062


78
V V H N S P S N D L
1
9063


92
V Q I S Y H S S H K
1
9064


106
A Y A V L Y L T C V
1
9065


108
A V L Y L T C V D I
1
9066


121
C D L N C E G R Q D
1
9067


125
C E G R Q D R N F V
1
9068


131
R N F V D K R Q W V
1
9069


132
N F V D K R Q W V W
1
9070


136
K R Q W V W G P S G
1
9071


141
W C P S G Y G G I L
1
9072


145
G Y G G I L L V N C
1
9073


147
G C I L L V N C D R
1
9074


149
I L L V N C D R D D
1
9075


152
V N C D R D D P S C
1
9076


157
D D P S C D V Q D N
1
9077


165
D N C D Q H V H C L
1
9078


171
V H C L Q D L K D M
1
9079


172
H C L Q D L K D M S
1
9080


173
C L Q D L E D M S V
1
9081


175
Q D L K D M S V M V
1
9082


182
V M V L R T Q G P A
1
9083


183
M V L R T Q G P A A
1
9084


198
K L V L H T S S Y D
1
9085


207
D A K R A Q V F H I
1
9086


211
A Q V F H I C G P E
1
9087


212
Q V F H I C C P E D
1
9088


213
V F H I C G P E D V
1
9089


215
H I C C P E D V C K
1
9090


227
R H V L G Q D K V S
1
9091


233
D K V S Y K V P R L
1
9092


238
K V P R L H C D E E
1
9093


239
V P R L H C D K E R
1
9094


242
L H G D E K R F F V
1
9095


249
F F V E G L S F P D
1
9096


251
V E G L S F P D A C
1
9097


253
G L S F P D A C F T
1
9098


259
A G F T G L I S F H
1
9099


260
G F T C L I S F H V
1
9100


267
F H V T L L D D S N
1
9101


283
P I F T D T V V F R
1
9102


284
I F T D T V V F R V
1
9103


286
T D T V V F R V A P
1
9104


289
V V F R V A P W I M
1
9105


291
F R V A P W I M T P
1
9106


293
V A P W I M T P S T
1
9107


294
A P W I M T P S T L
1
9108


304
P P L S V Y V C R V
1
9109


306
L E V Y V C R V R N
1
9110


307
E V Y V C R V R N N
1
9111


312
R V R N N T C F V D
1
9112


31
R N N T C F V D A V
1
9113


315
N N T C F V D A V A
1
9114


317
T C F V D A V A E L
1
9115


320
V D A V A S L A R K
1
9116


324
A E L A R K A G C K
1
9117


328
R K A G C K L T I C
1
9118


330
A G C K L T I C P Q
1
9119


333
K L T I C P Q A E N
1
9120


336
I C P Q A S N K N D
1
9121


337
C P Q A E N R N D R
1
9122


338
P Q A E N K N D R W
1
9123


344
N D R W I Q D E H E
1
9124


352
M S L G Y V Q A P H
1
9125


353
E L G Y V Q A P H K
1
9126


358
Q A P H K T L P V V
1
9127


360
P H K T L P V V F D
1
9128


361
H K T L P V V F D S
1
9129


363
T L P V V F D S P K
1
9130


371
P R N G S L Q D F P
1
9131


375
S L Q D F P Y K R I
1
9132


379
F P Y K K I L G P D
1
9133


380
P Y K R I L C P D F
1
9134


387
P D F G Y V T R I P
1
9135


388
D F G Y V T R S P R
1
9136


389
F G Y V T R E P R D
1
9137


395
E P R D R S V S G L
1
9138


404
L D S F C N L E V S
1
9139


407
F C N L E V S P P V
1
9140


408
G N L E V S P P V V
1
9141


410
L E V S P P V V A N
1
9142


416
V V A N G K E Y P L
1
9143


424
P L G R I L I G G N
1
9144


425
L G R I L I G G N L
1
9145


429
L I G G N L P C S S
1
9146


430
I C G N L P G S S C
1
9147


435
P G S S C R R V T Q
1
9148


439
G R R V T Q V V R D
1
9149


440
R R V T Q V V R D F
1
9150


441
R V T Q V V R D F L
1
9151


445
V V R D F L H A Q K
1
9152


447
R D F L H A Q K V Q
1
9153


451
H A Q K V Q P P V E
1
9154


452
A Q K V Q P P V E L
1
9155


457
P P V K L F V D W L
1
9156


466
L A V G H V D S F L
1
9157


472
D S F L S F V P A P
1
9158


479
P A P D G K G F R M
1
9159


486
F R H L L A S P G A
1
9160


489
L L A S P G A C F K
1
9161


498
K L F Q S K Q K C G
1
9162


501
Q S K Q K C G H G R
1
9163


503
K Q K C G H C R A L
1
9164


509
G R A L L F Q G V V
1
9165


513
L F Q C V V D D I Q
1
9166


514
F Q G V V D D E Q V
1
9167


515
Q G V V D D E Q V K
1
9168


528
I N Q V L S N K D L
1
9169


535
K D L I N Y N K F V
1
9170


538
I N Y N K F V Q S C
1
9171


543
F V Q S C I D W N K
1
9172


544
V Q S C I D W N R E
1
9173


557
R S L C L A S C D I
1
9174


558
E L G L A E C D I I
1
9175


573
F K T E R K K A T A
1
9176


591
L V L G K H L C I P
1
9177


595
K H L C I P K P F G
1
9178


596
H L G I P K P F G P
1
9179


598
C I P K P F G P I I
1
9180


601
K P F G P I I N G C
1
9181


609
C C C C L E S K V R
1
9182


611
C C L E S K V R S L
1
9183


622
E P L G L H C T F I
1
9184


623
P L G L H C T F I D
1
9185


625
G L H C T F I D D F
1
9186


638
H M L H G S V H C G
1
9187


642
G S V H C C T N V C
1
9188


643
S V H C C T N V C R
1
9189


646
C G T N V C R K P F
1
9190


654
P F S F K W W N H V
1
9191







184PEI2 v.2: HLA Peptide Scoring Results Al 10-mers


SYFPEITHI










6
P S T L A P L E V Y
20
9192


4
M T P S T L A P L E
8
9193


7
S T L A P L E V Y V
8
9194


8
T L A P L E V Y V C
7
9195


2
W I M T P S T L A P
6
9196


5
T P S T L A P L E V
6
9197


3
I M T P S T L A P L
4
9198


1
P W I M T P S T L A
2
9199


10
A P L E V Y V C R V
2
9200







184PIE2 v.3: HLA Peptide Scoring Results Al 10-mers


SYFPEITHI










10
V P D G K G F R M L
16
9201


1
V D E F L S F V P V
14
9202


6
S F V P V P D G K G
8
9203


8
V P V P D G K G F R
5
9204


5
L S F V P V P D G K
4
9205


4
F L S F V P V P D G
3
9206


2
D E F L S F V P V P
2
9207


7
F V P V P D G K G F
2
9208


9
P V P D G K G F R M
1
9209



















TABLE XXXVI








SEQ.





ID


Pos
1 2 3 4 5 6 7 8 9 0
score
NO.















184P1E2 v.1: HLA Peptide Scoring Results A0201 10-


mers SYFPEITHI










263
G L I S F H V T L L
27
9210


560
G L A E C D I I D I
27
9211


2
S L Q R I V R V S L
26
9212


297
I M T P S T L P P L
26
9213


192
A L F D D H K L V L
25
9214


184
V L R T Q G P A A L
24
9215


403
G L D S F G N L E V
24
9216


173
C L Q D L E D M S V
23
9217


176
D L E D M S V M V L
23
9218


17
A V C V A G V E T L
22
9219


301
S T L P P L E V Y V
22
9220


383
R I L G P D F G Y V
22
9221


428
I L I G G N L P G S
22
9222


433
N L P G S S G R R V
22
9223


520
L L E P L G L H C T
22
9224


325
E L A R K A G C K L
21
9225


612
C L E E K V R S L L
21
9226


280
S A S P I F T D T V
20
9227


490
L A S P G A C F K L
20
9228


566
I I D I P Q L F K T
20
9229


582
A F F P D L V N M L
20
9230


590
M L V L G K H L G I
20
9231


611
C C L E E K V R S L
20
9232


20
V A G V E T L V D I
19
9233


103
L P L A Y A V L Y L
19
9234


317
T C F V D A V A E L
19
9235


358
Q A P H K T L P V V
19
9236


384
I L G P D F G Y V T
19
9237


392
V T R S P R D R S V
19
9238


462
F V D W L A V G H V
19
9239


511
A L L F Q G V V D D
19
9240


517
V V D D E Q V K T I
19
9241


14
P T S A V C V A G V
18
9242


106
A Y A V L Y L T C V
18
9243


348
I Q D E M E L G Y V
18
9244


416
V V A N G K E Y P L
18
9245


454
K V Q P P V E L F V
18
9246


465
W L A V G H V D E F
18
9247


466
L A V G H V D E F L
18
9248


522
Q V K T I S I N Q V
18
9249


598
G I P K P F G P I I
18
9250


70
F D A T L E I I V V
17
9251


78
V V M N S P S N D L
17
9252


104
P L A Y A V L Y L T
17
9253


108
A V L Y L T C V D I
17
9254


302
T L P P L S V Y V C
17
9255


436
C S S G R R V T Q V
17
9256


512
L L F Q G V V D D E
17
9257


619
S L L E P L G L H C
17
9258


639
N L H C S V H C G T
17
9259


9
V S L E H P T S A V
16
9260


11
L E H P T S A V C V
16
9261


72
A T L E I I V V N N
16
9262


109
V L Y L T C V D I S
16
9263


110
L Y L T C V D I S L
16
9264


175
Q D L E D M S V H V
16
9265


190
P A A L F D D H K L
16
9266


191
A A L F D D H K L V
16
9267


204
S S Y D A K R A Q V
16
9268


221
D V C E A Y K H V L
16
9269


230
L G Q D K V S Y E V
16
9270


262
T C L I S F H V T L
16
9271


284
I F T D T V V F R V
16
9272


314
R N N T C F V D A V
16
9273


357
V Q A P H K T L P V
16
9274


408
G N L F V S P P V V
16
9275


469
C H V D S F L S F V
16
9276


571
Q L F K T E R K K A
16
9277


606
I I N G C C C L E E
16
9278


24
E T L V D I Y G S V
15
9279


28
D I Y G S V P E G T
15
9280


86
D L N D S H V Q I S
15
9281


111
Y L T C V D I S L D
15
9282


165
D N C D Q H V H C L
15
9283


229
V L G Q D K V S Y E
15
9284


294
A P W I M T P S T L
15
9285


304
P P L S V Y V C R V
15
9286


375
E L Q D F P Y K R I
15
9287


409
N L E V S P P V V A
15
9288


449
F L H A Q K V Q P P
15
9289


450
L H A Q K V Q P P V
15
9290


452
A Q K V Q P P V E L
15
9291


525
T I S I N Q V L S N
15
9292


531
V L S N K D L I N Y
15
9293


546
S C I D W N K S V L
15
9294


558
E L C L A E C D I I
15
9295


586
D L V N M L V L G K
15
9296


625
G L H C T F I D D F
15
9297


630
F I D D F T P Y H M
15
9298


638
H M L H C S V H C G
15
9299


16
S A V C V A C V E T
14
9300


33
V P E G T E M F E V
14
9301


36
C T E M F F V Y G T
14
9302


41
E V Y G T P G V D I
14
9303


65
T R R W R F D A T L
14
9304


68
W R F D A T L S I I
14
9305


114
C V D I S L D C D L
14
9306


118
S L D C D L N C E C
14
9307


149
I L L V N C D R D D
14
9308


162
D V Q D N C D Q H V
14
9309


207
D A K K A Q V F H I
14
9310


216
I C G P S D V C E A
14
9311


226
Y R H V L C Q D K V
14
9312


233
D K V S Y E V P R L
14
9313


253
C L S F P D A G F T
14
9314


255
S F P D A C F T G L
14
9315


333
K L T I C P Q A E N
14
9316


429
L I C C N L P G S S
14
9317


459
V E L F V D W L A V
14
9318


460
E L F V D W L A V C
14
9319


480
A P D G K G F K M L
14
9320


488
M L L A S P G A C F
14
9321


489
L L A S P C A C F K
14
9322


504
Q K C C H G R A L L
14
9323


516
G V V D D E Q V K T
14
9324


527
S I N Q V L S N K D
14
9325


550
W N R S V L K R E L
14
9326


578
K K A T A F F P D L
14
9327


579
K A T A F F P D L V
14
9328


581
T A F F P D L V N M
14
9329


591
L V L G K H L G I P
14
9330


597
L G I P K P F C P I
14
9331


10
S L E H P T S A V C
13
9332


51
Y I S P N M E R G R
13
9333


73
T L E I I V V M N S
13
9334


76
I I V V M N S P S N
13
9335


94
I S Y H S S H E P L
13
9336


96
Y H S S H E P L P L
13
9337


99
S H E P L P L A Y A
13
9338


101
E P L P L A Y A V L
13
9339


142
C P S C Y G C I L L
13
9340


168
D Q H V H C L Q D L
13
9341


179
D M S V M V L R T Q
13
9342


182
V M V L K T Q G P A
13
9343


199
L V L H T S S Y D A
13
9344


242
L H G D S E R F F V
13
9345


245
D E E R F F V E G L
13
9346


270
T L L D D S N E D F
13
9347


271
L L D D S N E D F S
13
9348


299
T P S T L P P L E V
13
9349


367
V F D S P R N G E L
13
9350


395
E P R D K S V S G L
13
9351


407
F G N L E V S P P V
13
9352


419
N G K S Y P L G R I
13
9353


441
R V T Q V V K D F L
13
9354


457
P P V S L F V D W L
13
9355


509
G R A L L F Q C V V
13
9356


587
L V N M L V L C K H
13
9357


604
G P I I N G C C C L
13
9358


617
V R S L L E P L G L
13
9359


635
T P Y H N L H G E V
13
9360


5
R I V R V S L S H P
12
9361


44
C T P C V D I Y I S
12
9362


71
D A T L E I I V V N
12
9363


85
N D L N D S H V Q I
12
9364


100
H E P L P L A Y A V
12
9365


140
V W G P S G Y G G I
12
9366


150
L L V N C D K D D P
12
9367


183
N V L K T Q C P A A
12
9368


200
V L H T S S Y D A K
12
9369


215
H I C C P E D V C E
12
9370


228
H V L G Q D K V S Y
12
9371


241
R L H G D E E R F F
12
9372


260
C F T G L I S F H V
12
9373


281
A S P I F T D T V V
12
9374


283
P I F T D T V V F R
12
9375


288
T V V F R V A P W I
12
9376


293
V A P W I M T P S T
12
9377


327
A R K A C C K L T I
12
9378


345
D R W I Q D E N E L
12
9379


355
G Y V Q A P H K T L
12
9380


421
K E Y P L G R I L I
12
9381


42
P L G R I L I G G N
12
9382


425
L G R I L I C G N L
12
9383


437
S S G R R V T Q V V
12
9384


446
V R D F L H A Q K V
12
9385


535
K D L I N Y N K F V
12
9386


537
L I N Y N K F V Q S
12
9387


545
Q S C I D W N R E V
12
9388


552
K E V L K R F L G L
12
9389


554
V L K R F L G L A F
12
9390


555
L K K E L G L A E C
12
9391


557
K E L C L A E C D I
12
9392


563
E C D I I D I P Q L
12
9393


583
F F P D L V N N L V
12
9394


584
F P D L V N M L V L
12
9395


608
N G C C C L E F K V
12
9396


631
I D D F T P Y H M L
12
9397


6
I V K V S L E H P T
11
9398


18
V C V A C V F T L V
11
9399


25
T L V D I Y G S V P
11
9400


39
M F E V Y G T P G V
11
9401


43
Y G T P C V D I Y I
11
9402


125
C E G K Q D R N F V
11
9403


131
R N F V D K K Q W V
11
9404


143
P S G Y C G I L L V
11
9405


148
G I L L V N C D R D
11
9406


154
C D R D D P S C D V
11
9407


198
K L V L H T S S Y D
11
9408


213
V F H I C G P F D V
11
9409


250
F V F G L S F P D A
11
9410


335
T I C P Q A E N R N
11
9411


339
Q A E N K N D K W I
11
9412


347
W I Q D E N E L G Y
11
9413


362
K T L P V V F D S P
11
9414


376
L Q D F P Y K R I L
11
9415


401
V S G L D S F C N L
11
9416


423
Y P L G R I L I G C
11
9417


427
R I L I G G N L P G
11
9418


47
F L S F V P A P D G
11
9419


487
R N L L A S P G A C
11
9420


498
K L F Q E K Q K C C
11
9421


503
K Q K C G H G R A L
11
9422


508
H G R A L L F Q G V
11
9423


514
F Q C V V D D E Q V
11
9424


528
I N Q V L S N K D L
11
9425


536
D L I N Y N K F V Q
11
9426


553
E V L K R E L G L A
11
9427


565
D I I D I P Q L F K
11
9428


588
V N M L V L G K H L
11
9429


589
N M L V L C K H L C
11
9430


592
V L G K H L C I P K
11
9431


596
H L G I P K P F G P
11
9432


615
E K V K S L L F P L
11
9433


8
K V S L E H P T S A
10
9434


47
C V D I Y I S P N M
10
9435


49
D I Y I S P N N E R
10
9436


67
R W K F D A T L E I
10
9437


69
R F D A T L F I I V
10
9438


79
V M N S P S N D L N
10
9439


93
Q I S Y H S S H E P
10
9440


105
L A Y A V L Y L T C
10
9441


117
I S L D C D L N C F
10
9442


122
D L N C F G R Q D K
10
9443


141
W G P S C Y C C I L
10
9444


144
S G Y G G I L L V N
10
9445


194
F D D H K L V L H T
10
9446


261
F T G L I S F H V T
10
9447


26
L I S F H V T L L D
10
9448


269
V T L L D D S N E D
10
9449


275
S N E D F S A S P I
10
9450


289
V V F R V A P W I M
10
9451


296
W I M T P S T L P P
10
9452


311
C R V R N N T C F V
10
9453


322
A V A F L A R K A G
10
9454


354
L G Y V Q A P H K T
10
9455


363
T L P V V F D S P R
10
9456


444
Q V V K D F L H A Q
10
9457


484
K G F K M L L A S P
10
9458


523
V K T I S I N Q V L
10
9459


547
C I D W N R E V L K
10
9460


548
I D W N R E V L K R
10
9461


573
F K T E R K K A T A
10
9462


601
K P F G P I I N G C
10
9463


634
F T P Y H M L H G E
10
9464


19
C V A G V E T L V D
9
9465


27
V D I Y G S V P E G
9
9466


83
P S N D L N D S H V
9
9467


97
H S S H F P L P L A
9
9468


102
P L P L A Y A V L Y
9
9469


139
W V W G P S G Y G G
9
9470


145
C Y C C I L L V N C
9
9471


171
V H C L Q D L E D M
9
9472


193
L F D D H K L V L H
9
9473


210
R A Q V F H I C G P
9
9474


256
F P D A G F T C L I
9
9475


285
F T D T V V F R V A
9
9476


287
D T V V F R V A P W
9
9477


309
Y V C R V R N N T C
9
9478


320
V D A V A E L A R K
9
9479


321
D A V A E L A R K A
9
9480


326
L A R K A G C K L T
9
9481


351
F H F L C Y V Q A P
9
9482


366
V V F D S P R N G E
9
9483


400
S V S G L D S F G N
9
9484


410
L E V S P P V V A N
9
9485


411
F V S P P V V A N G
9
9486


417
V A N C K F Y P L C
9
9487


420
C K E Y P L G R I L
9
9488


524
K T I S T N Q V L S
9
9489


538
I N Y N K F V Q S C
9
9490


539
N Y N K F V Q S C I
9
9491


561
L A F C D I I D I P
9
9492


568
D I P Q L F K T E R
9
9493


622
E P L C L H C T F I
9
9494


641
H G E V H C G T N V
9
9495


21
A G V E T L V D I Y
8
9496


26
L V D I Y C S V P E
8
9497


30
Y G S V P F C T E M
8
9498


32
S V P E C T E M F E
8
9499


38
E M F E V Y G T P G
8
9500


55
N H F R C R E R A D
8
9501


56
N E R C R E R A D T
8
9502


64
D T R R W R F D A T
8
9503


116
D I S L D C D L N C
8
9504


181
S V M V L R T Q G P
8
9505


212
Q V F H I C G P E D
8
9506


258
D A G F T G L I S F
8
9507


265
I S F H V T L L D D
8
9508


272
L D D S N E D F S A
8
9509


292
R V A P W I M T P S
8
9510


313
V R N N T C F V D A
8
9511


328
R K A G C K L T I C
8
9512


334
L T I C P Q A F N R
8
9513


350
D E N E L G Y V Q A
8
9514


445
V V R D F L H A Q K
8
9515


486
F R N L L A S P G A
8
9516


510
R A L L F Q G V V D
8
9517


519
D D E Q V K T I S I
8
9518


526
I S I N Q V L S N K
8
9519


529
N Q V L S N K D L I
8
9520


532
L S N K D L I N Y N
8
9521


580
A T A F F P D L V N
8
9522


593
L G K H L G I P K P
8
9523


607
I N G C C C L E E K
8
9524


74
L E I I V V M N S P
7
9525


75
F I I V V M N S P S
7
9526


112
L T C V D I S L D C
7
9527


133
F V D K R Q W V W G
7
9528


146
Y G C I L L V N C D
7
9529


151
L V N C D R D D P S
7
9530


170
H V H C L Q D L E D
7
9531


178
F D M S V N V L R T
7
9532


186
R T Q C P A A L F D
7
9533


202
H T S S Y D A K R A
7
9534


220
F D V C F A Y R H V
7
9535


223
C E A Y R H V L G Q
7
9536


236
S Y E V P R L H G D
7
9537


254
L S F P D A G F T G
7
9538


259
A G F T C L I S F H
7
9539


279
F S A S P I F T D T
7
9540


291
F R V A P W I N T P
7
9541


319
F V D A V A F L A R
7
9542


323
V A F L A R K A G C
7
9543


353
F L G Y V Q A P H K
7
9544


385
L G P D F G Y V T R
7
9545


439
G R R V T Q V V R D
7
9546


455
V Q P P V E L F V D
7
9547


481
P D G K G F R M L L
7
9548


506
C G H G R A L L F Q
7
9549


530
Q V L S N K D L I N
7
9550


567
I D I P Q L F K T F
7
9551


605
P I I N G C C C L E
7
9552


623
P L G L H C T F I D
7
9553


644
V H C G T N V C R K
7
9554


647
C T N V C R K P F S
7
9555


654
P F S F K W W N M V
7
9556


1
M S L Q R I V R V S
6
9557


3
L Q R I V R V S L E
6
9558


23
V E T L V D I Y G S
6
9559


77
I V V M N S P S N D
6
9560


91
H V Q I S Y H S S H
6
9561


98
S S H E P L P L A Y
6
9562


107
Y A V L Y L T C V D
6
9563


128
R Q D R N F V D K R
6
9564


174
L Q D L E D M S V M
6
9565


187
T Q G P A A L F D D
6
9566


195
D D H K L V L H T S
6
9567


206
Y D A K R A Q V F H
6
9568


224
E A Y R H V L G Q D
6
9569


244
C D E F R F F V E G
6
9570


266
S F H V T L L D D S
6
9571


274
D S N F D F S A S P
6
9572


305
P L F V Y V C R V R
6
9573


307
F V Y V C R V R N N
6
9574


308
V Y V C R V R N N T
6
9575


316
N T C F V D A V A E
6
9576


330
A G C K L T I C P Q
6
9577


342
N R N D R W I Q D F
6
9578


343
R N D R W I Q D E M
6
9579


352
M E L G Y V Q A P H
6
9580


356
Y V Q A P H K T L P
6
9581


370
S P K N C E L Q D F
6
9582


374
C E L Q D F P Y R K
6
9583


379
F P Y K R I L G P D
6
9584


402
S G L D S F C N L E
6
9585


405
D S F G N L E V S P
6
9586


406
S F G N L F V S P P
6
9587


434
L P C S S G R R V T
6
9588


461
L F V D W L A V G H
6
9589


467
A V G H V D E F L S
6
9590


470
H V D E F L S F V P
6
9591


472
D E F L S F V P A P
6
9592


476
S F V P A P D G K C
6
9593


479
P A P D C K G F R M
6
9594


483
G K G F R M L L A S
6
9595


494
C A C F K L F Q E K
6
9596


513
L F Q G V V D D E Q
6
9597


595
K H L G I P K P F C
6
9598


616
K V R S L L F P L C
6
9599


618
R S L L E P L G L H
6
9600


626
L H C T F I D D F T
6
9601


640
L H G E V H C G T N
6
9602


653
K P F S F K W W N M
6
9603


4
Q R I V R V S L E H
5
9604


12
E H P T S A V C V A
5
9605


42
V Y G T P G V D I Y
5
9606


53
S P N M E R G R E R
5
9607


63
A D T R R W R F D A
5
9608


81
N S P S N D L N D S
5
9609


90
S H V Q I S Y H S S
5
9610


113
T C V D I S L D C D
5
9611


134
V D K R Q W V W G P
5
9612


152
V N C D R D D P S C
5
9613


201
L H T S S Y D A K R
5
9614


235
V S Y E V P R L H G
5
9615


248
R F F V F C L S F P
5
9616


278
D F S A S P I F T D
5
9617


282
S P I F T D T V V F
5
9618


290
V F R V A P W I M T
5
9619


298
M T P S T L P P L E
5
9620


315
N N T C F V D A V A
5
9621


318
C F V D A V A E L A
5
9622


329
K A G C K L T I C P
5
9623


331
G C K L T I C P Q A
5
9624


391
Y V T R F P R D R S
5
9625


398
D R S V S G L D S F
5
9626


40
L D S F G N L F V S
5
9627


412
V S P P V V A N G K
5
9628


432
C N L P C S S C R R
5
9629


442
V T Q V V R D F L H
5
9630


443
T Q V V K D F L H A
5
9631


451
H A Q K V Q P P V F
5
9632


471
V D E F L S F V P A
5
9633


477
F V P A P D G K G F
5
9634


478
V P A P D C K G F K
5
9635


482
D G K C F R M L L A
5
9636


495
A C F K L F Q E K Q
5
9637


499
L F Q E K Q K C G H
5
9638


507
C H C R A L L F Q C
5
9639


542
K F V Q S C I D W N
5
9640


543
F V Q S C I D W N R
5
9641


572
L F K T E R K K A T
5
9642


574
K T F R K K A T A F
5
9643


585
P D L V N N L V L G
5
9644


628
C T F I D D F T P Y
5
9645


637
Y H M L H G E V H C
5
9646


7
V R V S L E H P T S
4
9647


22
G V E T L V D T Y G
4
9648


29
I Y G S V P E G T E
4
9649


50
I Y I S P N M E R G
4
9650


54
P N M E R C R H R A
4
9651


62
R A D T R R W R F D
4
9652


82
S P S N D L N D S H
4
9653


87
L N D S H V Q I S Y
4
9654


88
N D S H V Q I S Y H
4
9655


119
L D C D L N C E G R
4
9656


123
L N C H C R Q D R N
4
9657


127
C R Q D R N F V D K
4
9658


136
K R Q W V W G P S C
4
9659


156
R D D P S C D V Q D
4
9660


177
L E D M S V M V L R
4
9661


197
H K L V L H T S S Y
4
9662


234
K V S Y E V P R L H
4
9663


237
Y E V P R L H G D E
4
9664


303
L P P L H V Y V C R
4
9665


312
R V R N N T C F V D
4
9666


324
A E L A R K A G C K
4
9667


359
A P H K T L P V V F
4
9668


378
D F P Y K R I L G P
4
9669


382
K R I L C P D F G Y
4
9670


386
G P D F C Y V T R H
4
9671


394
R E P R D R S V S C
4
9672


413
S P P V V A N G K H
4
9673


418
A N G K E Y P L G R
4
9674


426
C R I L I G C N L P
4
9675


430
I G C N L P G S S G
4
9676


438
S G R R V T Q V V R
4
9677


440
R R V T Q V V R D F
4
9678


453
Q K V Q P P V E L F
4
9679


458
P V E L F V D W L A
4
9680


468
V G H V D E F L S F
4
9681


475
L S F V P A P D G K
4
9682


497
F K L F Q E K Q K C
4
9683


559
L G L A E C D I I D
4
9684


621
L E P L G L H C T F
4
9685


624
L G L H C T F I D D
4
9686


629
T F I D D F T P Y H
4
9687


649
N V C R K P F S F K
4
9688


31
C S V P E C T E M F
3
9689


48
V D I Y I S P N M E
3
9690


80
N N S P S N D L N D
3
9691


84
S N D L N D S H V Q
3
9692


92
V Q I S Y H S S H E
3
9693


95
S Y H S S H E P L P
3
9694


115
V D I S L D C D L N
3
9695


147
G G I L L V N C D R
3
9696


157
D D P S C D V Q D N
3
9697


164
Q D N C D Q H V H C
3
9698


185
L R T Q C P A A L F
3
9699


189
C P A A L F D D H K
3
9700


208
A K R A Q V F H I C
3
9701


209
K R A Q V F H I C C
3
9702


21
F H I C G P E D V C
3
9703


217
C G P E D V C E A Y
3
9704


218
C P H D V C E A Y R
3
9705


238
E V P R L H C D E E
3
9706


239
V P R L H G D E E R
3
9707


249
F F V E G L S F P D
3
9708


267
F H V T L L D D S N
3
9709


268
H V T L L D D S N H
3
9710


306
L E V Y V C R V R N
3
9711


332
C K L T I C P Q A E
3
9712


340
A E N R N D R W I Q
3
9713


361
H K T L P V V F D S
3
9714


364
L P V V F D S P R N
3
9715


368
F D S P R N G H L Q
3
9716


377
Q D F P Y K R I L G
3
9717


389
F G Y V T R E P R D
3
9718


431
C G N L P C S S G R
3
9719


448
D F L H A Q K V Q P
3
9720


463
V D W L A V G H V D
3
9721


464
D W L A V G H V D E
3
9722


491
A S P C A C F K L F
3
9723


492
S P G A C F K L F Q
3
9724


493
P G A C F K L F Q E
3
9725


518
V D D E Q V K T I S
3
9726


520
D E Q V K T I S I N
3
9727


533
S N K D L I N Y N K
3
9728


534
N K D L I N Y N K F
3
9729


549
D W N R E V L K R E
3
9730


562
A E C D I I D I P Q
3
9731


599
I P K P F G P I I N
3
9732


614
H E K V R S L L E P
3
9733


643
E V H C G T N V C R
3
9734


650
V C R K P F S F K W
3
9735


13
H P T S A V C V A G
2
9736


15
T S A V C V A G V E
2
9737


37
T E M F E V Y G T P
2
9738


45
T P C V D I Y I S P
2
9739


46
P G V D I Y I S P N
2
9740


52
I S P N M E R G R E
2
9741


58
R G R E R A D T R R
2
9742


121
C D L N C E G R Q D
2
9743


137
R Q W V W G P S G Y
2
9744


160
S C D V Q D N C D Q
2
9745


163
V Q D N C D Q H V H
2
9746


172
H C L Q D L E D N S
2
9747


205
S Y D A K R A Q V F
2
9748


222
V C H A Y R H V L G
2
9749


225
A Y R H V L G Q D K
2
9750


243
H G D E E R F F V E
2
9751


251
V E G L S F P D A G
2
9752


252
H G L S F P D A G F
2
9753


257
P D A G F T C L I S
2
9754


277
H D F S A S P I F T
2
9755


336
I C P Q A H N K N D
2
9757


346
K W I Q D H M E L G
2
9758


360
P H K T L P V V F D
2
9759


372
R N G H L Q D F P Y
2
9760


381
Y K R I L C P D F G
2
9761


387
P D F C Y V T K E P
2
9762


422
E Y P L G R I L I C
2
9763


456
Q P P V H L F V D W
2
9764


505
K C C H G R A L L F
2
9765


544
V Q S C I D W N R H
2
9766


564
C D I I D I P Q L F
2
9767


569
I P Q L F K T E R K
2
9768


575
T E R K K A T A F F
2
9769


610
C C C L E E K V R S
2
9770


613
L E H K V R S L L E
2
9771


642
C E V H C C T N V C
2
9772


645
H C G T N V C K K P
2
9773


648
T N V C K K P F S F
2
9774


655
F S F K W W N M V P
2
9775


66
R R W K F D A T L H
1
9776


124
N C H C R Q D R N F
1
9777


132
N F V D K K Q W V W
1
9778


167
C D Q H V H C L Q D
1
9779


211
A Q V F H I C C P E
1
9780


231
G Q D K V S Y E V P
1
9781


232
Q D K V S Y E V P R
1
9782


310
V C R V R N N T C F
1
9783


337
C P Q A E N R N D R
1
9784


338
P Q A E N R N D R W
1
9785


390
G Y V T R E P R D R
1
9786


393
T R H P R D R S V S
1
9787


397
R D R S V S G L D S
1
9788


415
P V V A N G K H Y P
1
9789


435
P G S S G R R V T Q
1
9790


473
E F L S F V P A P D
1
9791


485
G F R M L L A S P G
1
9792


501
Q E K Q K C G H G R
1
9793


502
E K Q K C G H G R A
1
9794


541
N K F V Q S C I D W
1
9795


603
F G P I I N G C C C
1
9796


633
D F T P Y H M L H G
1
9797


57
H R G R E R A D T R
−1
9798


120
D C D L N C E G R Q
−1
9799


129
Q D R N F V D K R Q
−1
9800


130
D R N F V D K R Q W
−1
9801


158
D P S C D V Q D N C
−1
9803


159
P S C D V Q D N C D
−1
9804


227
R H V L G Q D K V S
−1
9805


240
P R L H C D H E R F
−1
9806


295
P W I M T P S T L P
−1
9807


551
N R E V L K R E L C
−1
9808


577
K K K A T A F F P D
−1
9809


602
P F G P I I N C C C
−1
9810


652
K K P F S F K W W N
−1
9811


219
P E D V C E A Y R H
−2
9812


388
D F C Y V T R E P R
−2
9813


414
P P V V A N G K E Y
−2
9814


496
C F K L F Q E K Q K
−2
9815


646
C G T N V C R K P F
−2
9816


34
P E G T E M F H V Y
−3
9817


126
E G K Q D R N F V D
−3
9818


341
E N R N D K W I Q D
−3
9819


380
P Y K R I L C P D F
−3
9820


396
P R D R S V S G L D
−3
9821


521
E Q V K T I S I N Q
−3
9822


636
P Y H N L H C H V H
−3
9823


246
E E R F F V E G L S
−4
9824


576
E R K K A T A F F P
−4
9825







184P1E2 v.2: HLA Peptide Scoring Results A0201 10-


mers SYFPEAITHI










3
I M T P S T L A P L
27
9826


7
S T L A P L E V Y V
20
9827


10
A P L E V Y V C R V
20
9828


8
T L A P L E V Y V C
19
9829


5
T P S T L A P L E V
13
9830


2
W I M T P S T L A P
11
9831


9
L A P L E V Y V C R
8
9832


4
M T P S T L A P L E
5
9833


1
P W I M T P S T L A
3
9834







184P1E2 v.3: HLA Peptide Scoring Results A0201 10-


mers SYFPEAITHI










4
F L S F V P V P D G
15



10
V P D G K G F R M L
13



1
V D E F L S F V P V
11



6
S F V P V P D G K G
6



9
P V P D G K G F R M
6



2
D E F L S F V P V P
5



7
F V P V P D G K G F
5



5
L S F V P V P D G K
4



8
V P V P D G K G F R
3



3
E F L S F V P V P D
1




















TABLE XXXVII









SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A0202 10-mers SYFPEITHI












190
P A A L F D D H K L
5
9835



106
A Y A V L Y L T C V
4
9836



322
A V A E L A R K A G
4
9837



580
A T A F F P D L V N
4
9838



15
T S A V C V A G V E
3
9839



19
C V A G V E T L V D
3
9840



61
E R A D T R R W R F
3
9841



70
F D A T L E I I V V
3
9842



104
P L A Y A V L Y T L
3
9843



189
G P A A L F D D H K
3
9844



191
A A L F D D H K L V
3
9845



206
Y D A K R A Q V F H
3
9846



209
K R A Q V F H I C G
3
9847



223
C E A Y R H V L G Q
3
9848



257
P D A G F T G L I S
3
9849



279
F S A S P I F T D T
3
9850



292
R V A P W I M T P S
3
9851



320
V D A V A E L A R K
3
9852



325
E L A R K A G C K L
3
9853



328
R K A G C K L T I C
3
9854



338
P Q A E N R N D R W
3
9855



357
V Q A P H K T L P V
3
9856



416
V V A N G K E Y P L
3
9857



450
L H A Q K V Q P P V
3
9858



465
W L A V G H V D E F
3
9859



478
V P A P D G K G F R
3
9860



489
L L A S P G A C F K
3
9861



493
P G A C F K L F Q E
3
9862



509
G R A L L F Q G V V
3
9863



560
G L A E C D I I D I
3
9864



578
K K A T A F F P D L
3
9865



16
S A V C V A G V E T
2
9866



20
V A G V E T L V D I
2
9867



62
R A D T R R W R F D
2
9868



71
D A T L E I I V V M
2
9869



105
L A Y A V L Y L T C
2
9870



107
Y A V L Y L T C V D
2
9871



207
D A K R A Q V F H I
2
9872



210
R A Q V F H I C G P
2
9873



224
E A Y R H V L G Q D
2
9874



258
D A G F T G L I S F
2
9875



280
S A S P I F T D T V
2
9876



293
V A P W I M T P S T
2
9877



321
D A V A E L A R K A
2
9878



323
V A E L A R K A G C
2
9879



326
L A R K A G C K L T
2
9880



329
K A G C K L T I C P
2
9881



339
Q A E N R N D R W I
2
9882



358
Q A P H K T L P V V
2
9883



417
V A N G K E Y P L G
2
9884



451
H A Q K V Q P P V E
2
9885



466
L A V G H V D E F L
2
9886



479
P A P D G K G F R M
2
9887



490
L A S P G A C F K L
2
9888



494
G A C F K L F Q E K
2
9889



510
R A L L F Q G V V D
2
9890



561
L A E C D I I D I P
2
9891



579
K A T A F F P D L V
2
9892



581
T A F F P D L V N M
2
9893



17
A V C V A G V E T L
1
9894



21
A G V E T L V D I Y
1
9895



63
A D T R R W R F D A
1
9896



72
A T L E I I V V M N
1
9897



108
A V L Y L T C V D I
1
9898



192
A L F D D H K L V L
1
9899



208
A K R A Q V F H I C
1
9900



211
A Q V F H I C G P E
1
9901



225
A Y R H V L G Q D K
1
9902



259
A G F T G L I S F H
1
9903



281
A S P I F T D T V V
1
9904



294
A P W I M T P S T L
1
9905



324
A E L A R K A G C K
1
9906



327
A R K A G C K L T I
1
9907



330
A G C K L T I C P Q
1
9908



340
A E N R N D R W I Q
1
9909



359
A P H K T L P V V F
1
9910



418
A N G K E Y P L G R
1
9911



452
A Q K V Q P P V E L
1
9912



467
A V G H V D E F L S
1
9913



480
A P D G K G F R M L
1
9914



491
A S P G A C F K L F
1
9915



495
A C F K L F Q E K Q
1
9916



511
A L L F Q G V V D D
1
9917



562
A E C D I I D I P Q
1
9918



582
A F F P D L V N M L
1
9919







184P1E2 v.2: HLA Peptide


Scoring Results A0202 10-mers SYFPEITHI












8
T L A P L E V Y V C
3
9920



9
L A P L E V Y V C R
2
9921



10
A P L E V Y V C R V
1
9922




















TABLE XXXVIII









SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A0203 10-mers SYFPEITHI












183
M V L R T Q G P A A
19
9923



99
S H E P L P L A Y A
18
9924



315
N N T C F V D A V A
18
9925



573
F K T E R K K A T A
18
9926



184
V L R T Q G P A A L
17
9927



8
R V S L E H P T S A
10
9928



12
E H P T S A V C V A
10
9929



54
P N M E R G R E R A
10
9930



63
A D T R R W R F D A
10
9931



97
H S S H E P L P L A
10
9932



182
V M V L R T Q G P A
10
9933



199
L V L H T S S Y D A
10
9934



202
H T S S Y D A K R A
10
9935



216
I C G P E D V C E A
10
9936



250
F V E G L S F P D A
10
9937



272
L D D S N E D F S A
10
9938



285
F T D T V V F R V A
10
9939



313
V R N N T C F V D A
10
9940



318
C F V D A V A E L A
10
9941



321
D A V A E L A R K A
10
9942



331
G C K L T I C P Q A
10
9943



350
D E M E L G Y V Q A
10
9944



409
N L E V S P P V V A
10
9945



443
T Q V V R D F L H A
10
9946



458
P V E L F V D W L A
10
9947



471
V D E F L S F V P A
10
9948



482
D G K G F R M L L A
10
9949



486
F R M L L A S P G A
10
9950



502
E K Q K C G H G R A
10
9951



553
E V L K R E L G L A
10
9952



571
Q L F K T E R K K A
10
9953



9
V S L E H P T S A V
9
9954



13
H P T S A V C V A G
9
9955



55
N M E R G R E R A D
9
9956



64
D T R R W R F D A T
9
9957



98
S S H E P L P L A Y
9
9958



100
H E P L P L A Y A V
9
9959



200
V L H T S S Y D A K
9
9960



203
T S S Y D A K R A Q
9
9961



217
C G P E D V C E A Y
9
9962



251
V E G L S F P D A G
9
9963



273
D D S N E D F S A S
9
9964



286
T D T V V F R V A P
9
9965



314
R N N T C F V D A V
9
9966



316
N T C F V D A V A E
9
9967



319
F V D A V A E L A R
9
9968



322
A V A E L A R K A G
9
9969



332
C K L T I C P Q A E
9
9970



351
E M E L G Y V Q A P
9
9971



410
L E V S P P V V A N
9
9972



444
Q V V R D F L H A Q
9
9973



459
V E L F V D W L A V
9
9974



472
D E F L S F V P A P
9
9975



483
G K G F R M L L A S
9
9976



487
R M L L A S P G A C
9
9977



503
K Q K C G H G R A L
9
9978



554
V L K R E L G L A E
9
9979



572
L F K T E R K K A T
9
9980



574
K T E R K K A T A F
9
9981



10
S L E H P T S A V C
8
9982



14
P T S A V C V A G V
8
9983



56
M E R G R E R A D T
8
9984



65
T R R W R F D A T L
8
9985



101
E P L P L A Y A V L
8
9986



185
L R T Q G P A A L F
8
9987



201
L H T S S Y D A K R
8
9988



204
S S Y D A K R A Q V
8
9989



218
G P E D V C E A Y R
8
9990



252
E G L S F P D A G F
8
9991



274
D S N E D F S A S P
8
9992



287
D T V V F R V A P W
8
9993



317
T C F V D A V A E L
8
9994



320
V D A V A E L A R K
8
9995



323
V A E L A R K A G C
8
9996



333
K L T I C P Q A E N
8
9997



352
M E L G Y V Q A P H
8
9998



411
E V S P P V V A N G
8
9999



445
V V R D F L H A Q K
8
10000



460
E L F V D W L A V G
8
10001



473
E F L S F V P A P D
8
10002



484
K G F R M L L A S P
8
10003



488
M L L A S P G A C F
8
10004



504
Q K C G H G R A L L
8
10005



555
L K R E L G L A E C
8
10006



575
T E R K K A T A F F
8
10007







184P1E2 v.2: HLA Peptide


Scoring Results A0203 10-mers SYFPEITHI












1
P W I M T P S T L A
10
10008



2
W I M T P S T L A P
9
10009



3
I M T P S T L A P L
8
10010




















TABLE XXXIX









SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A3 10-mers SYFPEITHI












445
V V R D F L H A Q K
30
10011



228
H V L G Q D K V S Y
26
10012



649
N V C R K P F S F K
26
10013



565
D I I D I P Q L F K
25
10014



102
P L P L A Y A V L Y
24
10015



488
M L L A S P G A C F
24
10016



489
L L A S P G A C F K
24
10017



586
D L V N M L V L G K
24
10018



41
E V Y G T P G V D I
23
10019



353
E L G Y V Q A P H K
23
10020



592
V L G K H L G I P K
23
10021



312
R V R N N T C F V D
22
10022



384
I L G P D F G Y V T
22
10023



547
C I D W N R E V L K
22
10024



2
S L Q R I V R V S L
21
10025



25
T L V D I Y G S V P
21
10026



192
A L F D D H K L V L
21
10027



200
V L H T S S Y D A K
21
10028



324
A E L A R K A G C K
21
10029



554
V L K R E L G L A E
21
10030



19
C V A G V E T L V D
20
10031



91
H V Q I S Y H S S H
20
10032



241
R L H G D E E R F F
20
10033



319
F V D A V A E L A R
20
10034



619
S L L E P L G L H C
20
10035



8
R V S L E H P T S A
19
10036



108
A V L Y L T C V D I
19
10037



183
M V L R T Q G P A A
19
10038



225
A Y R H V L G Q D K
19
10039



292
R V A P W I M T P S
19
10040



409
N L E V S P P V V A
19
10041



438
S G R R V T Q V V R
19
10042



454
K V Q P P V E L F V
19
10043



477
F V P A P D G K G F
19
10044



511
A L L F Q G V V D D
19
10045



526
I S I N Q V L S N K
19
10046



10
S L E H P T S A V C
18
10047



49
D I Y I S P N M E R
18
10048



184
V L R T Q G P A A L
18
10049



234
K V S Y E V P R L H
18
10050



305
P L E V Y V C R V R
18
10051



347
W I Q D E M E L G Y
18
10052



427
R I L I G G N L P G
18
10053



510
R A L L F Q G V V D
18
10054



536
D L I N Y N K F V Q
18
10055



643
E V H C G T N V C R
18
10056



17
A V C V A G V E T L
17
10057



198
K L V L H T S S Y D
17
10058



270
T L L D D S N E D F
17
10059



302
T L P P L E V Y V C
17
10060



309
Y V C R V R N N T C
17
10061



322
A V A E L A R K A G
17
10062



325
E L A R K A G C K L
17
10063



333
K L T I C P Q A E N
17
10064



363
T L P V V F D S P R
17
10065



383
R I L G P D F G Y V
17
10066



428
I L I G G N L P G S
17
10067



460
E L F V D W L A V G
17
10068



530
Q V L S N K D L I N
17
10069



531
V L S N K D L I N Y
17
10070



77
I V V M N S P S N D
16
10071



176
D L E D M S V M V L
16
10072



221
D V C E A Y R H V L
16
10073



238
E V P R L H G D E E
16
10074



282
S P I F T D T V V F
16
10075



283
P I F T D T V V F R
16
10076



359
A P H K T L P V V F
16
10077



411
E V S P P V V A N G
16
10078



412
V S P P V V A N G K
16
10079



444
Q V V R D F L H A Q
16
10080



515
Q G V V D D E Q V K
16
10081



522
Q V K T I S I N Q V
16
10082



616
K V R S L L E P L G
16
10083



5
R I V R V S L E H P
15
10084



6
I V R V S L E H P T
15
10085



26
L V D I Y G S V P E
15
10086



28
D I Y G S V P E G T
15
10087



51
Y I S P N M E R G R
15
10088



60
R E R A D T R R W R
15
10089



109
V L Y L T C V D I S
15
10090



122
D L N C E G R Q D R
15
10091



127
G R Q D R N F V D K
15
10092



133
F V D K R Q W V W G
15
10093



139
W V W G P S G Y G G
15
10094



212
Q V F H I C G P E D
15
10095



433
N L P G S S G R R V
15
10096



441
R V T Q V V R D F L
15
10097



465
W L A V G H V D E F
15
10098



470
H V D E F L S F V P
15
10099



496
C F K L F Q E K Q K
15
10100



516
G V V D D E Q V K T
15
10101



517
V V D D E Q V K T I
15
10102



537
L I N Y N K F V Q S
15
10103



574
K T E R K K A T A F
15
10104



587
L V N M L V L G K H
15
10105



591
L V L G K H L G I P
15
10106



606
I I N G C C C L E E
15
10107



612
C L E E K V R S L L
15
10108



57
E R G R E R A D T R
14
10109



75
E I I V V M N S P S
14
10110



137
R Q W V W G P S G Y
14
10111



144
S G Y G G I L L V N
14
10112



149
I L L V N C D R D D
14
10113



156
R D D P S C D V Q D
14
10114



173
C L Q D L E D M S V
14
10115



205
S Y D A K R A Q V F
14
10116



215
H I C G P E D V C E
14
10117



253
G L S F P D A G F T
14
10118



263
G L I S F H V T L L
14
10119



268
H V T L L D D S N E
14
10120



288
T V V F R V A P W I
14
10121



289
V V F R V A P W I M
14
10122



307
E V Y V C R V R N N
14
10123



320
V D A V A E L A R K
14
10124



391
Y V T R E P R D R S
14
10125



424
P L G R I L I G G N
14
10126



429
L I G G N L P G S S
14
10127



525
T I S I N Q V L S N
14
10128



543
F V Q S C I D W N R
14
10129



590
M L V L G K H L G I
14
10130



625
G L H C T F I D D F
14
10131



32
S V P E G T E M F E
13
10132



58
R G R E R A D T R R
13
10133



98
S S H E P L P L A Y
13
10134



105
L A Y A V L Y L T C
13
10135



111
Y L T C V D I S L D
13
10136



170
H V H C L Q D L E D
13
10137



189
G P A A L F D D H K
13
10138



197
H K L V L H T S S Y
13
10139



204
S S Y D A K R A Q V
13
10140



229
V L G Q D K V S Y E
13
10141



327
A R K A G C K L T I
13
10142



356
Y V Q A P H K T L P
13
10143



382
K R I L G P D F G Y
13
10144



385
L G P D F G Y V T R
13
10145



394
R E P R D R S V S G
13
10146



397
R D R S V S G L D S
13
10147



400
S V S G L D S F G N
13
10148



403
G L D S F G N L E V
13
10149



467
A V G H V D E F L S
13
10150



474
F L S F V P A P D G
13
10151



498
K L F Q E K Q K C G
13
10152



505
K C G H G R A L L F
13
10153



533
S N K D L I N Y N K
13
10154



553
E V L K R E L G L A
13
10155



568
D I P Q L F K T E R
13
10156



569
I P Q L F K T E R K
13
10157



571
Q L F K T E R K K A
13
10158



620
L L E P L G L H C T
13
10159



4
Q R I V R V S L E H
12
10160



73
T L E I I V V M N S
12
10161



76
I I V V M N S P S N
12
10162



101
E P L P L A Y A V L
12
10163



116
D I S L D C D L N C
12
10164



150
L L V N C D R D D P
12
10165



186
R T Q G P A A L F D
12
10166



199
L V L H T S S Y D A
12
10167



247
E R F F V E G L S F
12
10168



250
F V E G L S F P D A
12
10169



294
A P W I M T P S T L
12
10170



366
V V F D S P R N G E
12
10171



415
P V V A N G K E Y P
12
10172



416
V V A N G K E Y P L
12
10173



449
F L H A Q K V Q P P
12
10174



461
L F V D W L A V G H
12
10175



560
G L A E C D I I D I
12
10176



567
I D I P Q L F K T E
12
10177



570
P Q L F K T E R K K
12
10178



596
H L G I P K P F G P
12
10179



605
P I I N G C C C L E
12
10180



607
I N G C C C L E E K
12
10181



618
R S L L E P L G L H
12
10182



639
M L H G E V H C G T
12
10183



644
V H C G T N V C R K
12
10184



47
G V D I Y I S P N M
11
10185



66
R R W R F D A T L E
11
10186



72
A T L E I I V V M N
11
10187



78
V V M N S P S N D L
11
10188



86
D L N D S H V Q I S
11
10189



104
P L A Y A V L Y L T
11
10190



114
C V D I S L D C D L
11
10191



118
S L D C D L N C E G
11
10192



128
R Q D R N F V D K R
11
10193



148
G I L L V N C D R D
11
10194



162
D V Q D N C D Q H V
11
10195



181
S V M V L R T Q G P
11
10196



224
E A Y R H V L G Q D
11
10197



232
Q D K V S Y E V P R
11
10198



262
T G L I S F H V T L
11
10199



264
L I S F H V T L L D
11
10200



271
L L D D S N E D F S
11
10201



296
W I M T P S T L P P
11
10202



300
P S T L P P L E V Y
11
10203



335
T I C P Q A E N R N
11
10204



352
M E L G Y V Q A P H
11
10205



365
P V V F D S P R N G
11
10206



372
R N G E L Q D F P Y
11
10207



373
N G E L Q D F P Y K
11
10208



375
E L Q D F P Y K R I
11
10209



380
P Y K R I L G P D F
11
10210



393
T R E P R D R S V S
11
10211



418
A N G K E Y P L G R
11
10212



421
K E Y P L G R I L I
11
10213



430
I G G N L P G S S G
11
10214



458
P V E L F V D W L A
11
10215



462
F V D W L A V G H V
11
10216



475
L S F V P A P D G K
11
10217



484
K G F R M L L A S P
11
10218



558
E L G L A E C D I I
11
10219



566
I I D I P Q L F K T
11
10220



580
A T A F F P D L V N
11
10221



598
G I P K P F G P I I
11
10222



623
P L G L H C T F I D
11
10223



636
P Y H M L H G E V H
11
10224



11
L E H P T S A V C V
10
10225



21
A G V E T L V D I Y
10
10226



22
G V E T L V D I Y G
10
10227



34
P E G T E M F E V Y
10
10228



65
T R R W R F D A T L
10
10229



67
R W R F D A T L E I
10
10230



85
N D L N D S H V Q I
10
10231



93
Q I S Y H S S H E P
10
10232



151
L V N C D R D D P S
10
10233



175
Q D L E D M S V M V
10
10234



185
L R T Q G P A A L F
10
10235



254
L S F P D A G F T G
10
10236



259
A G F T G L I S F H
10
10237



301
S T L P P L E V Y V
10
10238



334
L T I C P Q A E N R
10
10239



362
K T L P V V F D S P
10
10240



432
G N L P G S S G R R
10
10241



487
R M L L A S P G A C
10
10242



494
G A C F K L F Q E K
10
10243



501
Q E K Q K C G H G R
10
10244



512
L L F Q G V V D D E
10
10245



524
K T I S I N Q V L S
10
10246



546
S C I D W N R E V L
10
10247



548
I D W N R E V L K R
10
10248



575
T E R K K A T A F F
10
10249



630
F I D D F T P Y H M
10
10250



15
T S A V C V A G V E
9
10251



56
M E R G R E R A D T
9
10252



71
D A T L E I I V V M
9
10253



121
C D L N C E G R Q D
9
10254



163
V Q D N C D Q H V H
9
10255



193
L F D D H K L V L H
9
10256



206
Y D A K R A Q V F H
9
10257



227
R H V L G Q D K V S
9
10258



235
V S Y E V P R L H G
9
10259



252
E G L S F P D A G F
9
10260



257
P D A G F T G L I S
9
10261



281
A S P I F T D T V V
9
10262



303
L P P L E V Y V C R
9
10263



349
Q D E M E L G Y V Q
9
10264



369
D S P R N G E L Q D
9
10265



370
S P R N G E L Q D F
9
10266



414
P P V V A N G K E Y
9
10267



435
P G S S G R R V T Q
9
10268



447
R D F L H A Q K V Q
9
10269



448
D F L H A Q K V Q P
9
10270



452
A Q K V Q P P V E L
9
10271



468
V G H V D E F L S F
9
10272



478
V P A P D G K G F R
9
10273



485
G F R M L L A S P G
9
10274



491
A S P G A C F K L F
9
10275



527
S I N Q V L S N K D
9
10276



538
I N Y N K F V Q S C
9
10277



564
C D I I D I P Q L F
9
10278



621
L E P L G L H C T F
9
10279



16
S A V C V A G V E T
8
10280



31
G S V P E G T E M F
8
10281



53
S P N M E R G R E R
8
10282



82
S P S N D L N D S H
8
10283



87
L N D S H V Q I S Y
8
10284



136
K R Q W V W G P S G
8
10285



147
G G I L L V N C D R
8
10286



174
L Q D L E D M S V M
8
10287



177
L E D M S V M V L R
8
10288



188
Q G P A A L F D D H
8
10289



208
A K R A Q V F H I C
8
10290



217
C G P E D V C E A Y
8
10291



218
G P E D V C E A Y R
8
10292



239
V P R L H G D E E R
8
10293



275
S N E D F S A S P I
8
10294



280
S A S P I F T D T V
8
10295



310
V C R V R N N T C F
8
10296



316
N T C F V D A V A E
8
10297



328
R K A G C K L T I C
8
10298



350
D E M E L G Y V Q A
8
10299



405
D S F G N L E V S P
8
10300



408
G N L E V S P P V V
8
10301



425
L G R I L I G G N L
8
10302



431
G G N L P G S S G R
8
10303



437
S S G R R V T Q V V
8
10304



440
R R V T Q V V R D F
8
10305



463
V D W L A V G H V D
8
10306



464
D W L A V G H V D E
8
10307



503
K Q K C G H G R A L
8
10308



504
Q K C G H G R A L L
8
10309



552
R E V L K R E L G L
8
10310



557
R E L G L A E C D I
8
10311



573
F K T E R K K A T A
8
10312



584
F P D L V N M L V L
8
10313



609
G C C C L E E K V R
8
10314



628
C T F I D D F T P Y
8
10315



629
T F I D D F T P Y H
8
10316



3
L Q R I V R V S L E
7
10317



9
V S L E H P T S A V
7
10318



29
I Y G S V P E G T E
7
10319



61
E R A D T R R W R F
7
10320



63
A D T R R W R F D A
7
10321



64
D T R R W R F D A T
7
10322



88
N D S H V Q I S Y H
7
10323



99
S H E P L P L A Y A
7
10324



103
L P L A Y A V L Y L
7
10325



126
E G R Q D R N F V D
7
10326



153
N C D R D D P S C D
7
10327



161
C D V Q D N C D Q H
7
10328



201
L H T S S Y D A K R
7
10329



214
F H I C G P E D V C
7
10330



219
P E D V C E A Y R H
7
10331



248
R F F V E G L S F P
7
10332



291
F R V A P W I M T P
7
10333



315
N N T C F V D A V A
7
10334



337
C P Q A E N R N D R
7
10335



340
A E N R N D R W I Q
7
10336



341
E N R N D R W I Q D
7
10337



343
R N D K W I Q D E M
7
10338



346
R W I Q D E M E L G
7
10339



357
V Q A P H K T L P V
7
10340



374
G E L Q D F P Y K R
7
10341



395
E P R D R S V S G L
7
10342



399
R S V S G L D S F G
7
10343



402
S G L D S F G N L E
7
10344



426
G R I L I G G N L P
7
10345



439
G R R V T Q V V R D
7
10346



455
V Q P P V E L F V D
7
10347



509
G R A L L F Q G V V
7
10348



555
L K R E L G L A E C
7
10349



556
K R E L G L A E C D
7
10350



577
R K K A T A F F P D
7
10351



582
A F F P D L V N M L
7
10352



595
K H L G I P K P F G
7
10353



597
L G I P K P F G P I
7
10354



599
I P K P F G P I I N
7
10355



648
T N V C R K P F S F
7
10356



1
M S L Q R I V R V S
6
10357



40
F E V Y G T P G V D
6
10358



42
V Y G T P G V D I Y
6
10359



69
R F D A T L E I I V
6
10360



74
L E I I V V M N S P
6
10361



80
M N S P S N D L N D
6
10362



84
S N D L N D S H V Q
6
10363



92
V Q I S Y H S S H E
6
10364



94
I S Y H S S H E P L
6
10365



124
N C E G R Q D R N F
6
10366



129
Q D R N F V D K R Q
6
10367



131
R N F V D K R Q W V
6
10368



145
G Y G G I L L V N C
6
10369



164
Q D N C D Q H V H C
6
10370



167
C D Q H V H C L Q D
6
10371



222
V C E A Y R H V L G
6
10372



240
P R L H G D E E R F
6
10373



258
D A G F T G L I S F
6
10374



274
D S N E D F S A S P
6
10375



278
D F S A S P I F T D
6
10376



286
T D T V V F R V A P
6
10377



358
Q A P H K T L P V V
6
10378



368
F D S P R N G E L Q
6
10379



377
Q D F P Y K R I L G
6
10380



392
V T R E P R D R S V
6
10381



398
D R S V S G L D S F
6
10382



436
G S S G R R V T Q V
6
10383



453
Q K V Q P P V E L F
6
10384



456
Q P P V E L F V D W
6
10385



459
V E L F V D W L A V
6
10386



473
E F L S F V P A P D
6
10387



480
A P D G K G F R M L
6
10388



482
D G K G F R M L L A
6
10389



492
S P G A C F K L F Q
6
10390



507
G H G R A L L F Q G
6
10391



535
K D L I N Y N K F V
6
10392



579
K A T A F F P D L V
6
10393



585
P D L V N M L V L G
6
10394



594
G K H L G I P K P F
6
10395



633
D F T P Y H M L H G
6
10396



640
L H G E V H C G T N
6
10397



646
C G T N V C R K P F
6
10398



13
H P T S A V C V A G
5
10399



14
P T S A V C V A G V
5
10400



24
E T L V D I Y G S V
5
10401



35
E G T E M F E V Y G
5
10402



37
T E M F E V Y G T P
5
10403



38
E M F E V Y G T P G
5
10404



48
V D I Y I S P N M E
5
10405



50
I Y I S P N M E R G
5
10406



52
I S P N M E R G R E
5
10407



70
F D A T L E I I V V
5
10408



100
H E P L P L A Y A V
5
10409



106
A Y A V L Y L T C V
5
10410



107
Y A V L Y L T C V D
5
10411



117
I S L D C D L N C E
5
10412



119
L D C D L N C E G R
5
10413



132
N F V D K R Q W V W
5
10414



142
G P S G Y G G I L L
5
10415



155
D R D D P S C D V Q
5
10416



180
M S V M V L R T Q G
5
10417



191
A A L F D D H K L V
5
10418



196
D H K L V L H T S S
5
10419



216
I C G P E D V C E A
5
10420



243
H G D E E R F F V E
5
10421



244
G D E E R F F V E G
5
10422



261
F T G L I S F H V T
5
10423



265
I S F H V T L L D D
5
10424



273
D D S N E D F S A S
5
10425



297
I M T P S T L P P L
5
10426



306
L E V Y V C R V R N
5
10427



313
V R N N T C F V D A
5
10428



314
R N N T C F V D A V
5
10429



321
D A V A E L A R K A
5
10430



331
G C K L T I C P Q A
5
10431



354
L G Y V Q A P H K T
5
10432



360
P H K T L P V V F D
5
10433



379
F P Y K R I L G P D
5
10434



381
Y K R I L G P D F G
5
10435



388
D F G Y V T R E P R
5
10436



390
G Y V T R E P R D R
5
10437



406
S F G N L E V S P P
5
10438



410
L E V S P P V V A N
5
10439



434
L P G S S G R R V T
5
10440



451
H A Q K V Q P P V E
5
10441



476
S F V P A P D G K G
5
10442



499
L F Q E K Q K C G H
5
10443



506
C G H G K A L L F Q
5
10444



508
H G R A L L F Q G V
5
10445



523
V K T I S I N Q V L
5
10446



534
N K D L I N Y N K F
5
10447



601
K P F G P I I N G C
5
10448



610
C C C L E E K V R S
5
10449



611
C C L E E K V R S L
5
10450



617
V R S L L E P L G L
5
10451



622
E P L G L H C T F I
5
10452



632
D D F T P Y H M L H
5
10453



635
T P Y H M L H G E V
5
10454



637
Y H M L H G E V H C
5
10455



651
C R K P F S F K W W
5
10456



652
R K P F S F K W W N
5
10457



655
F S F K W W N M V P
5
10458



7
V R V S L E H P T S
4
10459



12
E H P T S A V C V A
4
10460



20
V A G V E T L V D I
4
10461



30
Y G S V P E G T E M
4
10462



46
P G V D I Y I S P N
4
10463



55
N M E R G R E R A D
4
10464



62
R A D T R R W R F D
4
10465



83
P S N D L N D S H V
4
10466



96
Y H S S H E P L P L
4
10467



110
L Y L T C V D I S L
4
10468



125
C E G K Q D R N F V
4
10469



134
V D K R Q W V W G P
4
10470



135
D K R Q W V W G P S
4
10471



140
V W G P S G Y G G I
4
10472



141
W G P S G Y G G I L
4
10473



154
C D R D D P S C D V
4
10474



179
D M S V M V L R T Q
4
10475



195
D D H K L V L H T S
4
10476



207
D A K R A Q V F H I
4
10477



209
K R A Q V F H I C G
4
10478



210
R A Q V F H I C G P
4
10479



211
A Q V F H I C G P E
4
10480



223
C E A Y R H V L G Q
4
10481



231
G Q D K V S Y E V P
4
10482



237
Y E V P R L H G D E
4
10483



246
E E R F F V E G L S
4
10484



276
N E D F S A S P I F
4
10485



285
F T D T V V F R V A
4
10486



290
V F R V A P W I M T
4
10487



295
P W I M T P S T L P
4
10488



299
T P S T L P P L E V
4
10489



318
C F V D A V A E L A
4
10490



323
V A E L A R K A G C
4
10491



332
C K L T I C P Q A E
4
10492



339
Q A E N R N D R W I
4
10493



344
N D R W I Q D E M E
4
10494



348
I Q D E M E L G Y V
4
10495



355
G Y V Q A P H K T L
4
10496



389
F G Y V T R E P R D
4
10497



396
P R D R S V S G L D
4
10498



404
L D S F G N L E V S
4
10499



422
E Y P L G R I L I G
4
10500



423
Y P L G R I L I G C
4
10501



442
V T Q V V R D F L H
4
10502



469
G H V D E F L S F V
4
10503



471
V D E F L S F V P A
4
10504



479
P A P D G K C F R M
4
10505



483
G K G F R M L L A S
4
10506



493
P G A C F K L F Q E
4
10507



542
K F V Q S C I D W N
4
10508



550
W N R E V L K R E L
4
10509



559
L G L A E C D I I D
4
10510



562
A E C D I I D I P Q
4
10511



563
E C D I I D I P Q L
4
10512



576
E R K K A T A F F P
4
10513



578
K K A T A F F P D L
4
10514



581
T A F F P D L V N M
4
10515



588
V N M L V L G K H L
4
10516



602
P F G P I I N G C C
4
10517



603
F G P I I N G C C C
4
10518



604
G P I I N G C C C L
4
10519



614
E E K V R S L L E P
4
10520



627
H C T F I D D F T P
4
10521



641
H G E V H C G T N V
4
10522



642
G E V H C G T N V C
4
10523



18
V C V A G V E T L V
3
10524



27
V D I Y G S V P E G
3
10525



43
Y G T P G V D I Y I
3
10526



59
G R E R A D T R R W
3
10527



90
S H V Q I S Y H S S
3
10528



112
L T C V D I S L D C
3
10529



115
V D I S L D C D L N
3
10530



130
D R N F V D K R Q W
3
10531



138
Q W V W G P S G Y G
3
10532



143
P S G Y G G I L L V
3
10533



152
V N C D R D D P S C
3
10534



172
H C L Q D L E D M S
3
10535



178
E D M S V M V L R T
3
10536



187
T Q G P A A L F D D
3
10537



194
F D D H K L V L H T
3
10538



202
H T S S Y D A K R A
3
10539



245
D E E R F F V E G L
3
10540



251
V E G L S F P D A G
3
10541



255
S F P D A G F T G L
3
10542



256
F P D A G F T G L I
3
10543



269
V T L L D D S N E D
3
10544



284
I F T D T V V F R V
3
10545



287
D T V V F R V A P W
3
10546



304
P P L E V Y V C R V
3
10547



311
C R V R N N T C F V
3
10548



317
T C F V D A V A E L
3
10549



326
L A R K A G C K L T
3
10550



329
K A G C K L T I C P
3
10551



330
A G C K L T I C P Q
3
10552



378
D F P Y K R I L G P
3
10553



419
N G K E Y P L G R I
3
10554



420
G K E Y P L G R I L
3
10555



443
T Q V V R D F L H A
3
10556



446
V R D F L H A Q K V
3
10557



490
L A S P G A C F K L
3
10558



495
A C F K L F Q E K Q
3
10559



514
F Q G V V D D E Q V
3
10560



520
D E Q V K T I S I N
3
10561



539
N Y N K F V Q S C I
3
10562



545
Q S C I D W N R E V
3
10563



549
D W N R F V L K R E
3
10564



613
L E E K V R S L L E
3
10565



615
E K V R S L L E P L
3
10566



631
I D D F T P Y H M L
3
10567



650
V C R K P F S F K W
3
10568



653
K P F S F K W W N M
3
10569



33
V P E G T E M F E V
2
10570



44
G T P G V D I Y I S
2
10571



45
T P G V D I Y I S P
2
10572



54
S N M E R G R E R A
2
10573



120
D C D L N C E G R Q
2
10574



169
Q H V H C L Q D L E
2
10575



182
V M V L R T Q G P A
2
10576



190
P A A L F D D H K L
2
10577



213
V F H I C G P E D V
2
10578



242
L H G D E E R F F V
2
10579



260
G F T G L I S F H V
2
10580



267
F H V T L L D D S N
2
10581



272
L D D S N E D F S A
2
10582



279
F S A S P I F T D T
2
10583



293
V A P W I M T P S T
2
10584



336
I C P Q A E N R N D
2
10585



338
P Q A E N R N D R W
2
10586



345
D R W I Q D E M E L
2
10587



351
E M E L G Y V Q A P
2
10588



371
P R N G E L Q D F P
2
10589



387
P D F G Y V T R E P
2
10590



401
V S G L D S F G N L
2
10591



413
S P P V V A N G K E
2
10592



417
V A N G K E Y P L G
2
10593



457
P P V E L F V D W L
2
10594



466
L A V G H V D E F L
2
10595



472
D E F L S F V P A P
2
10596



481
P D G K G F R M L L
2
10597



497
F K L F Q E K Q K C
2
10598



513
L F Q G V V D D E Q
2
10599



518
V D D E Q V K T I S
2
10600



519
D D E Q V K T I S I
2
10601



528
I N Q V L S N K D L
2
10602



572
L F K T E R K K A T
2
10603



583
F F P D L V N M L V
2
10604



589
N M L V L G K H L G
2
10605



624
L G L H C T F I D D
2
10606



638
H M L H G E V H C G
2
10607



23
V E T L V D I Y G S
1
10608



68
W R F D A T L E I I
1
10609



95
S Y H S S H E P L P
1
10610



113
T C V D I S L D C D
1
10611



123
L N C E G R Q D R N
1
10612



146
Y G G I L L V N C D
1
10613



159
P S C D V Q D N C D
1
10614



160
S C D V Q D N C D Q
1
10615



165
D N C D Q H V H C L
1
10616



171
V H C L Q D L E D M
1
10617



203
T S S Y D A K R A Q
1
10618



230
L G Q D K V S Y E V
1
10619



236
S Y E V P R L H G D
1
10620



249
F F V E G L S F P D
1
10621



266
S F H V T L L D D S
1
10622



277
E D F S A S P I F T
1
10623



298
M T P S T L P P L E
1
10624



308
V Y V C R V R N N T
1
10625



342
N R N D R W I Q D E
1
10626



361
H K T L P V V F D S
1
10627



367
V F D S P R N G E L
1
10628



386
G P D F G Y V T R E
1
10629



407
F G N L E V S P P V
1
10630



450
L H A Q K V Q P P V
1
10631



486
F R M L L A S P G A
1
10632



500
F Q E K Q K C G H G
1
10633



502
E K Q K C G H G R A
1
10634



529
N Q V L S N K D L I
1
10635



540
Y N K F V Q S C I D
1
10636



551
N R E V L K R E L G
1
10637



593
L G K H L G I P K P
1
10638



654
P F S F K W W N M V
1
10639







184P1E2 v.2: HLA Peptide


Scoring Results A3 10-mers SYFPEITHI












8
T L A P L E V Y V C
18
10640



2
W I M T P S T L A P
12
10641



6
P S T L A P L E V Y
11
10642



7
S T L A P L E V Y V
9
10643



9
L A P L E V Y V C R
9
10644



3
I M T P S T L A P L
8
10645



10
A P L E V Y V C R V
6
10646



1
P W I M T P S T L A
4
10647



5
T P S T L A P L E V
4
10648



4
M T P S T L A P L E
3
10649







184P1E2 v.3: HLA Peptide


Scoring Results A3 10-mers SYFPEITHI












7
F V P V P D G K G F
17
10650



9
P V P D G K G F R M
14
10651



5
L S F V P V P D G K
11
10652



4
F L S F V P V P D G
10
10653



8
V P V P D G K G F R
8
10654



3
E F L S F V P V P D
7
10655



2
D E F L S F V P V P
5
10656



6
S F V P V P D G K G
5
10657



1
V D E F L S F V P V
4
10658



10
V P D G K G F R M L
3
10659




















TABLE XL









SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results A26 10-mers SYFPEITHI












176
D L E D M S V M V L
28
10660



221
D V C E A Y R H V L
26
10661



628
C T F I D D F T P Y
26
10662



228
H V L G Q D K V S Y
24
10663



325
E L A R K A G C K L
24
10664



411
E V S P P V V A N G
24
10665



465
W L A V G H V D E F
24
10666



263
G L I S F H V T L L
23
10667



531
V L S N K D L I N Y
23
10668



582
A F F P D L V N M L
23
10669



17
A V C V A G V E T L
22
10670



347
W I Q D E M E L G Y
22
10671



477
F V P A P D G K G F
22
10672



625
G L H C T F I D D F
22
10673



24
E T L V D I Y G S V
21
10674



47
G V D I Y I S P N M
21
10675



64
D T R R W R F D A T
21
10676



101
E P L P L A Y A V L
21
10677



233
D K V S Y E V P R L
21
10678



255
S F P D A G F T G L
21
10679



258
D A G F T G L I S F
21
10680



287
D T V V F R V A P W
21
10681



307
E V Y V C R V R N N
21
10682



395
E P R D R S V S G L
21
10683



398
D R S V S G L D S F
21
10684



460
E L F V D W L A V G
21
10685



553
E V L K R E L G L A
21
10686



563
E C D I I D I P Q L
21
10687



574
K T E R K K A T A F
21
10688



630
F I D D F T P Y H M
21
10689



86
D L N D S H V Q I S
20
10690



102
P L P L A Y A V L Y
20
10691



241
R L H G D E E R F F
20
10692



289
V V F R V A P W I M
20
10693



375
E L Q D F P Y K R I
20
10694



28
D I Y G S V P E G T
19
10695



41
E V Y G T P G V D I
19
10696



61
E R A D T R R W R F
19
10697



71
D A T L E I I V V M
19
10698



75
E I I V V M N S P S
19
10699



114
C V D I S L D C D L
19
10700



162
D V Q D N C D Q H V
19
10701



165
D N C D Q H V H C L
19
10702



168
D Q H V H C L Q D L
19
10703



192
A L F D D H K L V L
19
10704



245
D E E R F F V E G L
19
10705



247
E R F F V E G L S F
19
10706



252
E G L S F P D A G F
19
10707



270
T L L D D S N E D F
19
10708



416
V V A N G K E Y P L
19
10709



441
R V T Q V V R D F L
19
10710



488
M L L A S P G A C F
19
10711



565
D I I D I P Q L F K
19
10712



568
D I P Q L F K T E R
19
10713



586
D L V N M L V L G K
19
10714



612
C L E E K V R S L L
19
10715



633
D F T P Y H M L H G
19
10716



2
S L Q R I V R V S L
18
10717



49
D I Y I S P N M E R
18
10718



72
A T L E I I V V M N
18
10719



78
V V M N S P S N D L
18
10720



184
V L R T Q G P A A L
18
10721



238
E V P R L H G D E E
18
10722



378
D F P Y K R I L G P
18
10723



428
I L I G G N L P G S
18
10724



615
E K V R S L L E P L
18
10725



44
G T P G V D I Y I S
17
10726



116
D I S L D C D L N C
17
10727



250
F V E G L S F P D A
17
10728



283
P I F T D T V V F R
17
10729



292
R V A P W I M T P S
17
10730



362
K T L P V V F D S P
17
10731



367
V F D S P R N G E L
17
10732



491
A S P G A C F K L F
17
10733



643
E V H C G T N V C R
17
10734



14
P T S A V C V A G V
16
10735



21
A G V E T L V D I Y
16
10736



278
D F S A S P I F T D
16
10737



317
T C F V D A V A E L
16
10738



345
D R W I Q D E M E L
16
10739



353
E L G Y V Q A P H K
16
10740



383
R I L G P D F G Y V
16
10741



462
F V D W L A V G H V
16
10742



480
A P D G K G F R M L
16
10743



517
V V D D E Q V K T I
16
10744



522
Q V K T I S I N Q V
16
10745



534
N K D L I N Y N K F
16
10746



536
D L I N Y N K F V Q
16
10747



537
L I N Y N K F V Q S
16
10748



566
I I D I P Q L F K T
16
10749



581
T A F F P D L V N M
16
10750



591
L V L G K H L G I P
16
10751



634
F T P Y H M L H G E
16
10752



5
R I V R V S L E H P
15
10753



36
G T E M F E V Y G T
15
10754



122
D L N C E G R Q D R
15
10755



285
F T D T V V F R V A
15
10756



297
I M T P S T L P P L
15
10757



302
T L P P L E V Y V C
15
10758



370
S P R N G E L Q D F
15
10759



444
Q V V R D F L H A Q
15
10760



453
Q K V Q P P V E L F
15
10761



468
V G H V D E F L S F
15
10762



512
L L F Q G V V D D E
15
10763



525
T I S I N Q V L S N
15
10764



560
G L A E C D I I D I
15
10765



587
L V N M L V L G K H
15
10766



611
C C L E E K V R S L
15
10767



649
N V C R K P F S F K
15
10768



32
S V P E G T E M F E
14
10769



34
P E G T E M F E V Y
14
10770



42
V Y G T P G V D I Y
14
10771



73
T L E I I V V M N S
14
10772



174
L Q D L E D M S V M
14
10773



212
Q V F H I C G P E D
14
10774



217
C G P E D V C E A Y
14
10775



229
V L G Q D K V S Y E
14
10776



261
F T G L I S F H V T
14
10777



300
P S T L P P L E V Y
14
10778



322
A V A E L A R K A G
14
10779



350
D E M E L G Y V Q A
14
10780



366
V V F D S P R N G E
14
10781



440
R R V T Q V V R D F
14
10782



448
D F L H A Q K V Q P
14
10783



457
P P V E L F V D W L
14
10784



472
D E F L S F V P A P
14
10785



473
E F L S F V P A P D
14
10786



524
K T I S I N Q V L S
14
10787



558
E L G L A E C D I I
14
10788



578
K K A T A F F P D L
14
10789



620
L L E P L G L H C T
14
10790



631
I D D F T P Y H M L
14
10791



19
C V A G V E T L V D
13
10792



31
G S V P E G T E M F
13
10793



51
Y I S P N M E R G R
13
10794



87
L N D S H V Q I S Y
13
10795



98
S S H E P L P L A Y
13
10796



104
P L A Y A V L Y L T
13
10797



111
Y L T C V D I S L D
13
10798



124
N C E G R Q D R N F
13
10799



133
F V D K R Q W V W G
13
10800



139
W V W G P S G Y G G
13
10801



157
D D P S C D V Q D N
13
10802



193
L F D D H K L V L H
13
10803



200
V L H T S S Y D A K
13
10804



215
H I C G P E D V C E
13
10805



248
R F F V E G L S F P
13
10806



282
S P I F T D T V V F
13
10807



298
M T P S T L P P L E
13
10808



301
S T L P P L E V Y V
13
10809



334
L T I C P Q A E N R
13
10810



335
T I C P Q A E N R N
13
10811



382
K R I L G P D F G Y
13
10812



392
V T R E P R D R S V
13
10813



400
S V S G L D S F G N
13
10814



401
V S G L D S F G N L
13
10815



424
P L G K I L I G G N
13
10816



429
L I G G N L P G S S
13
10817



445
V V R D F L H A Q K
13
10818



449
F L H A Q K V Q P P
13
10819



470
H V D E F L S F V P
13
10820



482
D G K G F R M L L A
13
10821



511
A L L F Q G V V D D
13
10822



516
G V V D D E Q V K T
13
10823



564
C D I I D I P Q L F
13
10824



584
F P D L V N M L V L
13
10825



598
G I P K P F G P I I
13
10826



606
I I N G C C C L E E
13
10827



26
L V D I Y G S V P E
12
10828



76
I I V V M N S P S N
12
10829



77
I V V M N S P S N D
12
10830



91
H V Q I S Y H S S H
12
10831



103
L P L A Y A V L Y L
12
10832



109
V L Y L T C V D I S
12
10833



137
R Q W V W G P S G Y
12
10834



171
V H C L Q D L E D M
12
10835



178
E D M S V N V L R T
12
10836



181
S V M V L R T Q G P
12
10837



183
M V L R T Q G P A A
12
10838



185
L R T Q G P A A L F
12
10839



186
R T Q G P A A L F D
12
10840



197
H K L V L H T S S Y
12
10841



202
H T S S Y D A K R A
12
10842



205
S Y D A K R A Q V F
12
10843



207
D A K R A Q V F H I
12
10844



268
H V T L L D D S N E
12
10845



273
D D S N E D F S A S
12
10846



276
N E D F S A S P I F
12
10847



284
I F T D T V V F R V
12
10848



316
N T C F V D A V A E
12
10849



319
F V D A V A E L A R
12
10850



351
E M E L G Y V Q A P
12
10851



359
A P H K T L P V V F
12
10852



372
R N G E L Q D F P Y
12
10853



380
P Y K R I L G P D F
12
10854



388
D F G Y V T R E P R
12
10855



405
D S F G N L E V S P
12
10856



433
N L P G S S G R R V
12
10857



454
K V Q P P V E L F V
12
10858



527
S I N Q V L S N K D
12
10859



543
F V Q S C I D W N R
12
10860



547
C I D W N R E V L K
12
10861



571
Q L F K T E R K K A
12
10862



575
T E R K K A T A F F
12
10863



605
P I I N G C C C L E
12
10864



614
E E K V R S L L E P
12
10865



616
K V R S L L E P L G
12
10866



653
K P F S F K W W N M
12
10867



8
R V S L E H P T S A
11
10868



12
E H P T S A V C V A
11
10869



93
Q I S Y H S S H E P
11
10870



108
A V L Y L T C V D I
11
10871



148
G I L L V N C D R D
11
10872



170
H V H C L Q D L E D
11
10873



173
C L Q D L E D M S V
11
10874



195
D D H K L V L H T S
11
10875



220
E D V C E A Y R H V
11
10876



224
E A Y R H V L G Q D
11
10877



240
P R L H G D E E R F
11
10878



269
V T L L D D S N E D
11
10879



288
T V V F R V A P W I
11
10880



365
P V V F D S P R N G
11
10881



391
Y V T R E P R D R S
11
10882



403
G L D S F G N L E V
11
10883



406
S F G N L E V S P P
11
10884



414
P P V V A N G K E Y
11
10885



422
E Y P L G R I L I G
11
10886



452
A Q K V Q P P V E L
11
10887



479
P A P D G K G F R M
11
10888



490
L A S P G A C F K L
11
10889



498
K L F Q E K Q K C G
11
10890



504
Q K C G H G R A L L
11
10891



520
D E Q V K T I S I N
11
10892



530
Q V L S N K D L I N
11
10893



542
K F V Q S C I D W N
11
10894



546
S C I D W N R E V L
11
10895



549
D W N R E V L K R E
11
10896



550
W N R E V L K R E L
11
10897



580
A T A F F P D L V N
11
10898



619
S L L E P L G L H C
11
10899



621
L E P L G L H C T F
11
10900



639
M L H G E V H C G T
11
10901



647
G T N V C R K P F S
11
10902



648
T N V C R K P F S F
11
10903



6
I V R V S L E H P T
10
10904



22
G V E T L V D I Y G
10
10905



25
T L V D I Y G S V P
10
10906



38
E M F E V Y G T P G
10
10907



112
L T C V D I S L D C
10
10908



118
S L D C D L N C E G
10
10909



141
W G P S G Y G G I L
10
10910



142
G P S G Y G G I L L
10
10911



151
L V N C D R D D P S
10
10912



179
D M S V M V L R T Q
10
10913



190
P A A L F D D H K L
10
10914



199
L V L H T S S Y D A
10
10915



234
K V S Y E V P R L H
10
10916



264
L I S F H V T L L D
10
10917



266
S F H V T L L D D S
10
10918



271
L L D D S N E D F S
10
10919



274
D S N E D F S A S P
10
10920



296
W I M T P S T L P P
10
10921



309
Y V C R V R N N T C
10
10922



310
V C R V R N N T C F
10
10923



312
R V R N N T C F V D
10
10924



333
K L T I C P Q A E N
10
10925



343
R N D R W I Q D E M
10
10926



356
Y V Q A P H K T L P
10
10927



363
T L P V V F D S P R
10
10928



384
I L G P D F G Y V T
10
10929



415
P V V A N G K E Y P
10
10930



427
R I L I G G N L P G
10
10931



442
V T Q V V R D F L H
10
10932



458
P V E L F V D W L A
10
10933



466
L A V G H V D E F L
10
10934



467
A V G H V D E F L S
10
10935



481
P D G K G F R M L L
10
10936



489
L L A S P G A C F K
10
10937



503
K Q K C G H G R A L
10
10938



505
K C G H G R A L L F
10
10939



554
V L K R E L G L A E
10
10940



594
G K H L G I P K P F
10
10941



604
G P I I N G C C C L
10
10942



629
T F I D D F T P Y H
10
10943



646
C G T N V C R K P F
10
10944



10
S L E H P T S A V C
9
10945



30
Y G S V P E G T E M
9
10946



35
E G T E M F E V Y G
9
10947



65
T R R W R F D A T L
9
10948



96
Y H S S H E P L P L
9
10949



110
L Y L T C V D I S L
9
10950



198
K L V L H T S S Y D
9
10951



260
G F T G L I S F H V
9
10952



262
T G L I S F H V T L
9
10953



277
E D F S A S P I F T
9
10954



294
A P W I M T P S T L
9
10955



355
G Y V Q A P H K T L
9
10956



376
L Q D F P Y K R I L
9
10957



409
N L E V S P P V V A
9
10958



425
L G R I L I G G N L
9
10959



461
L F V D W L A V G H
9
10960



474
F L S F V P A P D G
9
10961



476
S F V P A P D G K G
9
10962



499
L F Q E K Q K C G H
9
10963



513
L F Q G V V D D E Q
9
10964



523
V K T I S I N Q V L
9
10965



552
R E V L K R E L G L
9
10966



583
F F P D L V N M L V
9
10967



588
V N M L V L G K H L
9
10968



590
M L V L G K H L G I
9
10969



592
V L G K H L G I P K
9
10970



596
H L G I P K P F G P
9
10971



601
K P F G P I I N G C
9
10972



617
V R S L L E P L G L
9
10973



632
D D F T P Y H M L H
9
10974



27
V D I Y G S V P E G
8
10975



39
M F E V Y G T P G V
8
10976



50
I Y I S P N M E R G
8
10977



94
I S Y H S S H E P L
8
10978



120
D C D L N C E G R Q
8
10979



145
G Y G G I L L V N C
8
10980



149
I L L V N C D R D D
8
10981



150
L L V N C D R D D P
8
10982



155
D R D D P S C D V Q
8
10983



196
D H K L V L H T S S
8
10984



244
G D E E R F F V E G
8
10985



249
F F V E G L S F P D
8
10986



253
G L S F P D A G F T
8
10987



305
P L E V Y V C R V R
8
10988



369
D S P R N G E L Q D
8
10989



419
N G K E Y P L G R I
8
10990



420
G K E Y P L G R I L
8
10991



484
K G F R M L L A S P
8
10992



496
C F K L F Q E K Q K
8
10993



502
E K Q K C G H G R A
8
10994



519
D D E Q V K T I S I
8
10995



521
E Q V K T I S I N Q
8
10996



526
I S I N Q V L S N K
8
10997



528
I N Q V L S N K D L
8
10998



555
L K R E L G L A E C
8
10999



567
I D I P Q L F K T E
8
11000



572
L F K T E R K K A T
8
11001



576
E R K K A T A F F P
8
11002



597
L G I P K P F G P I
8
11003



602
P F G P I I N G C C
8
11004



622
E P L G L H C T F I
8
11005



623
P L G L H C T F I D
8
11006



33
V P E G T E M F E V
7
11007



68
W R F D A T L E I I
7
11008



69
R F D A T L E I I V
7
11009



74
L E I I V V M N S P
7
11010



89
D S H V Q I S Y H S
7
11011



99
S H E P L P L A Y A
7
11012



126
E G R Q D R N F V D
7
11013



130
D R N F V D K R Q W
7
11014



132
N F V D K R Q W V W
7
11015



135
D K R Q W V W G P S
7
11016



140
V W G P S G Y G G I
7
11017



158
D P S C D V Q D N C
7
11018



216
I C G P E D V C E A
7
11019



243
H G D E E R F F V E
7
11020



259
A G F T G L I S F H
7
11021



290
V F R V A P W I M T
7
11022



304
P P L E V Y V C R V
7
11023



318
C F V D A V A E L A
7
11024



320
V D A V A E L A R K
7
11025



321
D A V A E L A R K A
7
11026



328
R K A G C K L T I C
7
11027



341
E N R N D R W I Q D
7
11028



346
R W I Q D E M E L G
7
11029



348
I Q D E M E L G Y V
7
11030



385
L G P D F G Y V T R
7
11031



410
L E V S P P V V A N
7
11032



417
V A N G K E Y P L G
7
11033



423
Y P L G R I L I G G
7
11034



436
G S S G R R V T Q V
7
11035



455
V Q P P V E L F V D
7
11036



464
D W L A V G H V D E
7
11037



485
G F R M L L A S P G
7
11038



593
L G K H L G I P K P
7
11039



644
V H C G T N V C R K
7
11040



651
C R K P F S F K W W
7
11041



654
P F S F K W W N M V
7
11042



46
P G V D I Y I S P N
6
11043



57
E R G R E R A D T R
6
11044



81
N S P S N D L N D S
6
11045



90
S H V Q I S Y H S S
6
11046



127
G R Q D R N F V D K
6
11047



128
R Q D R N F V D K R
6
11048



134
V D K R Q W V W G P
6
11049



144
S G Y G G I L L V N
6
11050



177
L E D M S V M V L R
6
11051



187
T Q G P A A L F D D
6
11052



194
F D D H K L V L H T
6
11053



213
V F H I C G P E D V
6
11054



223
C E A Y R H V L G Q
6
11055



231
G Q D K V S Y E V P
6
11056



246
E E R F F V E G L S
6
11057



265
I S F H V T L L D D
6
11058



279
F S A S P I F T D T
6
11059



303
L P P L E V Y V C R
6
11060



342
N R N D R W I Q D E
6
11061



358
Q A P H K T L P V V
6
11062



360
P H K T L P V V F D
6
11063



386
G P D F G Y V T R E
6
11064



439
G R R V T Q V V R D
6
11065



456
Q P P V E L F V D W
6
11066



469
G H V D E F L S F V
6
11067



493
P G A C F K L F Q E
6
11068



494
G A C F K L F Q E K
6
11069



506
C G H G R A L L F Q
6
11070



508
H G R A L L F Q G V
6
11071



538
I N Y N K F V Q S C
6
11072



600
P K P F G P I I N G
6
11073



1
M S L Q R I V R V S
5
11074



3
L Q R I V R V S L E
5
11075



20
V A G V E T L V D I
5
11076



23
V E T L V D I Y G S
5
11077



45
T P G V D I Y I S P
5
11078



70
F D A T L E I I V V
5
11079



88
N D S H V Q I S Y H
5
11080



92
V Q I S Y H S S H E
5
11081



97
H S S H E P L P L A
5
11082



106
A Y A V L Y L T C V
5
11083



113
T C V D I S L D C D
5
11084



117
I S L D C D L N C E
5
11085



156
R D D P S C D V Q D
5
11086



188
Q G P A A L F D D H
5
11087



210
R A Q V F H I C G P
5
11088



236
S Y E V P R L H G D
5
11089



291
F R V A P W I M T P
5
11090



313
V R N N T C F V D A
5
11091



314
R N N T C F V D A V
5
11092



330
A G C K L T I C P Q
5
11093



373
N G E L Q D F P Y K
5
11094



379
F P Y K R I L G P D
5
11095



387
P D F G Y V T R E P
5
11096



443
T Q V V R D F L H A
5
11097



471
V D E F L S F V P A
5
11098



483
G K G F R M L L A S
5
11099



507
G H G R A L L F Q G
5
11100



532
L S N K D L I N Y N
5
11101



548
I D W N K S V L K R
5
11102



577
R K K A T A F F P D
5
11103



585
P D L V N M L V L G
5
11104



607
I N G C C C L E E K
5
11105



9
V S L E H P T S A V
4
11106



37
T E M F E V Y G T P
4
11107



105
L A Y A V L Y L T C
4
11108



143
P S G Y G G I L L V
4
11109



146
Y G G I L L V N C D
4
11110



175
Q D L E D M S V M V
4
11111



208
A K R A Q V F H I C
4
11112



254
L S F P D A G F T G
4
11113



338
P Q A E N R N D R W
4
11114



361
H K T L P V V F D S
4
11115



377
Q D F P Y K R I L G
4
11116



404
L D S F G N L E V S
4
11117



412
V S P P V V A N G K
4
11118



495
A C F K L F Q E K Q
4
11119



561
L A E C D I I D I P
4
11120



618
R S L L E P L G L H
4
11121



624
L G L H C T F I D D
4
11122



638
H M L H G E V H C G
4
11123



655
F S F K W W N M V P
4
11124



4
Q R I V R V S L E H
3
11125



11
L E H P T S A V C V
3
11126



13
H P T S A V C V A G
3
11127



48
V D I Y I S P N M E
3
11128



54
P N M E R G R E R A
3
11129



56
M E R G R E R A D T
3
11130



58
R G R E R A D T R R
3
11131



80
M N S P S N D L N D
3
11132



83
P S N D L N D S H V
3
11133



115
V D I S L D C D L N
3
11134



123
L N C E G R Q D R N
3
11135



131
R N F V D K R Q W V
3
11136



152
V N C D R D D P S C
3
11137



201
L H T S S Y D A K R
3
11138



204
S S Y D A K R A Q V
3
11139



206
Y D A K R A Q V F H
3
11140



235
V S Y E V P R L H G
3
11141



257
P D A G F T G L I S
3
11142



272
L D D S N E D F S A
3
11143



293
V A P W I M T P S T
3
11144



331
G C K L T I C P Q A
3
11145



336
I C P Q A E N R N D
3
11146



352
M E L G Y V Q A P H
3
11147



357
V Q A P H K T L P V
3
11148



393
T R E P R D R S V S
3
11149



394
R E P R D R S V S G
3
11150



408
G N L S V S P P V V
3
11151



432
G N L P G S S G R R
3
11152



446
V R D F L H A Q K V
3
11153



450
L H A Q K V Q P P V
3
11154



475
L S F V P A P D G K
3
11155



478
V P A P D G K G F R
3
11156



501
Q E K Q K C G H G R
3
11157



518
V D D E Q V K T I S
3
11158



533
S N K D L I N Y N K
3
11159



573
F K T E R K K A T A
3
11160



595
K H L G I P K P F G
3
11161



599
I P K P F G P I I N
3
11162



7
V R V S L E H P T S
2
11163



16
S A V C V A G V E T
2
11164



43
Y G T P G V D I Y I
2
11165



52
I S P N M E R G R E
2
11166



85
N D L N D S H V Q I
2
11167



95
S Y H S S H E P L P
2
11168



100
H E P L P L A Y A V
2
11169



119
L D C D L N C E G R
2
11170



136
K R Q W V W G P S G
2
11171



147
G G I L L V N C D R
2
11172



153
N C D R D D P S C D
2
11173



161
C D V Q D N C D Q H
2
11174



164
Q D N C D Q H V H C
2
11175



166
N C D Q H V H C L Q
2
11176



172
H C L Q D L E D M S
2
11177



189
G P A A L F D D H K
2
11178



209
K K A Q V F H I C G
2
11179



214
F H I C G P E D V C
2
11180



218
G P E D V C E A Y R
2
11181



219
P E D V C E A Y R H
2
11182



230
L G Q D K V S Y E V
2
11183



232
Q D K V S Y E V P R
2
11184



237
Y E V P R L H G D E
2
11185



275
S N E D F S A S P I
2
11186



280
S A S P I F T D T V
2
11187



286
T D T V V F R V A P
2
11188



295
P W I M T P S T L P
2
11189



306
L E V Y V C R V R N
2
11190



323
V A E L A R K A G C
2
11191



332
C K L T I C P Q A E
2
11192



364
L P V V F D S P R N
2
11193



368
F D S P R N G E L Q
2
11194



371
P R N G E L Q D F P
2
11195



374
G E L Q D F P Y K R
2
11196



389
F G Y V T R E P R D
2
11197



399
R S V S G L D S F G
2
11198



413
S P P V V A N G K E
2
11199



418
A N G K E Y P L G R
2
11200



421
K E Y P L G R I L I
2
11201



426
G R I L I G G N L P
2
11202



430
I G G N L P G S S G
2
11203



431
G G N L P G S S G R
2
11204



435
P G S S G R R V T Q
2
11205



447
R D F L H A Q K V Q
2
11206



451
H A Q K V Q P P V E
2
11207



486
F R M L L A S P G A
2
11208



497
F K L F Q E K Q K C
2
11209



500
F Q E K Q K C G H G
2
11210



539
N Y N K F V Q S C I
2
11211



541
N K F V Q S C I D W
2
11212



557
R E L G L A E C D I
2
11213



610
C C C L E E K V R S
2
11214



637
Y H M L H G E V H C
2
11215



640
L H G E V H C G T N
2
11216



650
V C R K P F S F K W
2
11217



15
T S A V C V A G V E
1
11218



29
I Y G S V P E G T E
1
11219



53
S P N M E R G R E R
1
11220



55
N M E R G R E R A D
1
11221



59
G R E R A D T R R W
1
11222



62
R A D T R R W R F D
1
11223



67
R W R F D A T L E I
1
11224



79
V M N S P S N D L N
1
11225



82
S P S N D L N D S H
1
11226



84
S N D L N D S H V Q
1
11227



121
C D L N C E G R Q D
1
11228



129
Q D R N F V D K R Q
1
11229



154
C D R D D P S C D V
1
11230



159
P S C D V Q D N C D
1
11231



160
S C D V Q D N C D Q
1
11232



163
V Q D N C D Q H V H
1
11233



167
C D Q H V H C L Q D
1
11234



180
M S V M V L R T Q G
1
11235



203
T S S Y D A K R A Q
1
11236



222
V C E A Y R H V L G
1
11237



225
A Y R H V L G Q D K
1
11238



226
Y R H V L G Q D K V
1
11239



239
V P R L H G D E E R
1
11240



242
L H G D E E R F F V
1
11241



251
V E G L S F P D A G
1
11242



256
F P D A G F T G L I
1
11243



267
F H V T L L D D S N
1
11244



281
A S P I F T D T V V
1
11245



299
T P S T L P P L E V
1
11246



308
V Y V C R V R N N T
1
11247



315
N N T C F V D A V A
1
11248



326
L A R K A G C K L T
1
11249



327
A R K A G C K L T I
1
11250



344
N D R W I Q D E M E
1
11251



354
L G Y V Q A P H K T
1
11252



396
P R D R S V S G L D
1
11253



397
R D R S V S G L D S
1
11254



402
S G L D S F G N L E
1
11255



407
F G N L E V S P P V
1
11256



434
L P G S S G R R V T
1
11257



437
S S G R R V T Q V V
1
11258



438
S G R R V T Q V V R
1
11259



459
V E L F V D W L A V
1
11260



487
R M L L A S P G A C
1
11261



509
G R A L L F Q G V V
1
11262



510
R A L L F Q G V V D
1
11263



514
F Q G V V D D E Q V
1
11264



515
Q G V V D D E Q V K
1
11265



540
Y N K F V Q S C I D
1
11266



544
V Q S C I D W N R E
1
11267



545
Q S C I D W N R E V
1
11268



562
A E C D I I D I P Q
1
11269



569
I P Q L F K T E R K
1
11270



570
P Q L F K T E R K K
1
11271



603
F G P I I N G C C C
1
11272



608
N G C C C L E E K V
1
11273



613
L E E K V R S L L E
1
11274



635
T P Y H M L H G E V
1
11275



641
H G E V H C G T N V
1
11276



645
H C G T N V C R K P
1
11277



652
R K P F S F K W W N
1
11278







184P1E2 v.2: HLA Peptide


Scoring Results A26 10-mers SYFPEITHI












3
I M T P S T L A P L
15
11279



8
T L A P L E V Y V C
15
11280



6
P S T L A P L E V Y
14
11281



4
M T P S T L A P L E
13
11282



2
W I M T P S T L A P
11
11283



7
S T L A P L E V Y V
11
11284



10
A P L E V Y V C R V
7
11285



9
L A P L E V Y V C R
6
11286



1
P W I M T P S T L A
2
11287



5
T P S T L A P L E V
1
11288







184P1E2 v.3: HLA Peptide


Scoring Results A26 10-mers SYFPEITHI












7
F V P V P D G K G F
22
11289



9
P V P D G K G F R M
21
11290



10
V P D G K G F R M L
16
11291



3
E F L S F V P V P D
14
11292



2
D E F L S F V P V P
13
11293



4
F L S F V P V P D G
13
11294



6
S F V P V P D G K G
9
11295



1
V D E F L S F V P V
5
11296



5
L S F V P V P D G K
3
11297



8
V P V P D G K G F R
2
11298





















TABLE XLI










SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.
















184P1E2 v.1: HLA Peptide


Scoring Results B 0702 10-mers SYFPEITHI












480
A P D G K G F R M L
26
11299



101
E P L P L A Y A V L
24
11300



103
L P L A Y A V L Y L
24
11301



142
G P S G Y G G I L L
24
11302



395
E P R D R S V S G L
24
11303



584
F P D L V N M L V L
24
11304



294
A P W I M T P S T L
22
11305



359
A P H K T L P V V F
22
11306



457
P P V E L F V D W L
21
11307



622
E P L G L H C T F I
21
11308



282
S P I F T D T V V F
20
11309



299
T P S T L P P L E V
20
11310



604
G P I I N G C C C L
20
11311



256
F P D A G F T G L I
19
11312



434
L P G S S G R R V T
19
11313



33
V P E G T E M F E V
18
11314



370
S P R N G E L Q D F
18
11315



304
P P L E V Y V C R V
17
11316



653
K P F S F K W W N M
17
11317



635
T P Y H M L H G E V
16
11318



96
Y H S S H E P L P L
15
11319



192
A L F D D H K L V L
15
11320



492
S P G A C F K L F Q
15
11321



184
V L R T Q G P A A L
14
11322



297
I M T P S T L P P L
14
11323



452
A Q K V Q P P V E L
14
11324



504
Q K C G H G R A L L
14
11325



578
K K A T A F F P D L
14
11326



582
A F F P D L V N M L
14
11327



617
V R S L L E P L G L
14
11328



2
S L Q R I V R V S L
13
11329



13
H P T S A V C V A G
13
11330



17
A V C V A G V E T L
13
11331



65
T R R W R F D A T L
13
11332



386
G P D F G Y V T R E
13
11333



441
R V T Q V V R D F L
13
11334



456
Q P P V E L F V D W
13
11335



466
L A V G H V D E F L
13
11336



478
V P A P D G K G F R
13
11337



481
P D G K G F R M L L
13
11338



490
L A S P G A C F K L
13
11339



552
R E V L K R E L G L
13
11340



563
E C D I I D I P Q L
13
11341



601
K P F G P I I N G C
13
11342



56
M E R G R E R A D T
12
11343



67
R W R F D A T L E I
12
11344



82
S P S N D L N D S H
12
11345



94
I S Y H S S H E P L
12
11346



158
D P S C D V Q D N C
12
11347



176
D L E D M S V M V L
12
11348



189
G P A A L F D D H K
12
11349



221
D V C E A Y R H V L
12
11350



233
D K V S Y E V P R L
12
11351



239
V P R L H G D E E R
12
11352



255
S F P D A G F T G L
12
11353



262
T G L I S F H V T L
12
11354



263
G L I S F H V T L L
12
11355



303
L P P L E V Y V C R
12
11356



317
T C F V D A V A E L
12
11357



325
E L A R K A G C K L
12
11358



337
C P Q A E N R N D R
12
11359



357
V Q A P H K T L P V
12
11360



379
F P Y K R I L G P D
12
11361



401
V S G L D S F G N L
12
11362



416
V V A N G K E Y P L
12
11363



423
Y P L G R I L I G G
12
11364



425
L G R I L I G G N L
12
11365



436
G S S G R R V T Q V
12
11366



503
K Q K C G H G R A L
12
11367



546
S C I D W N R E V L
12
11368



550
W N R E V L K R E L
12
11369



569
I P Q L F K T E R K
12
11370



575
T E R K K A T A F F
12
11371



599
I P K P F G P I I N
12
11372



615
E K V R S L L E P L
12
11373



631
I D D F T P Y H M L
12
11374



14
P T S A V C V A G V
11
11375



41
E V Y G T P G V D I
11
11376



45
T P G V D I Y I S P
11
11377



78
V V M N S P S N D L
11
11378



114
C V D I S L D C D L
11
11379



165
D N C D Q H V H C L
11
11380



178
E D M S V M V L R T
11
11381



190
P A A L F D D H K L
11
11382



218
G P E D V C E A Y R
11
11383



245
D E E R F F V E G L
11
11384



327
A R K A G C K L T I
11
11385



364
L P V V F D S P R N
11
11386



367
V F D S P R N G E L
11
11387



376
L Q D F P Y K R I L
11
11388



454
K V Q P P V E L F V
11
11389



491
A S P G A C F K L F
11
11390



523
V K T I S I N Q V L
11
11391



528
I N Q V L S N K D L
11
11392



588
V N M L V L G K H L
11
11393



611
C C L E E K V R S L
11
11394



612
C L E E K V R S L L
11
11395



6
I V R V S L E H P T
10
11396



8
R V S L E H P T S A
10
11397



53
S P N M E R G R E R
10
11398



64
D T R R W R F D A T
10
11399



106
A Y A V L Y L T C V
10
11400



108
A V L Y L T C V D I
10
11401



110
L Y L T C V D I S L
10
11402



141
W G P S G Y G G I L
10
11403



168
D Q H V H C L Q D L
10
11404



194
F D D H K L V L H T
10
11405



216
I C G P E D V C E A
10
11406



241
R L H G D E E R F F
10
11407



250
F V E G L S F P D A
10
11408



280
S A S P I F T D T V
10
11409



281
A S P I F T D T V V
10
11410



345
D R W I Q D E M E L
10
11411



355
G Y V Q A P H K T L
10
11412



383
R I L G P D F G Y V
10
11413



384
I L G P D F G Y V T
10
11414



413
S P P V V A N G K E
10
11415



414
P P V V A N G K E Y
10
11416



420
G K E Y P L G R I L
10
11417



471
V D E F L S F V P A
10
11418



482
D G K G F R M L L A
10
11419



505
K C G H G R A L L F
10
11420



508
H G R A L L F Q G V
10
11421



11
L E H P T S A V C V
9
11422



12
E H P T S A V C V A
9
11423



20
V A G V E T L V D I
9
11424



30
Y G S V P E G T E M
9
11425



61
E R A D T R R W R F
9
11426



63
A D T R R W R F D A
9
11427



69
R F D A T L E I I V
9
11428



71
D A T L E I I V V M
9
11429



97
H S S H E P L P L A
9
11430



125
C E G R Q D R N F V
9
11431



143
P S G Y G G I L L V
9
11432



205
S Y D A K R A Q V F
9
11433



242
L H G D E E R F F V
9
11434



247
E R F F V E G L S F
9
11435



253
G L S F P D A G F T
9
11436



261
F T G L I S F H V T
9
11437



279
F S A S P I F T D T
9
11438



285
F T D T V V F R V A
9
11439



301
S T L P P L E V Y V
9
11440



310
V C R V R N N T C F
9
11441



313
V R N N T C F V D A
9
11442



314
R N N T C F V D A V
9
11443



326
L A R K A G C K L T
9
11444



350
D E M E L G Y V Q A
9
11445



403
G L D S F G N L E V
9
11446



409
N L E V S P P V V A
9
11447



421
K E Y P L G R I L I
9
11448



437
S S G R R V T Q V V
9
11449



443
T Q V V R D F L H A
9
11450



450
L H A Q K V Q P P V
9
11451



459
V E L F V D W L A V
9
11452



468
V G H V D E F L S F
9
11453



557
R E L G L A E C D I
9
11454



574
K T E R K K A T A F
9
11455



597
L G I P K P F G P I
9
11456



9
V S L E H P T S A V
8
11457



16
S A V C V A G V E T
8
11458



18
V C V A G V E T L V
8
11459



28
D I Y G S V P E G T
8
11460



31
G S V P E G T E M F
8
11461



39
M F E V Y G T P G V
8
11462



47
G V D I Y I S P N M
8
11463



54
P N M E R G R E R A
8
11464



70
F D A T L E I I V V
8
11465



85
N D L N D S H V Q I
8
11466



99
S H E P L P L A Y A
8
11467



124
N C E G R Q D R N F
8
11468



131
R N F V D K R Q W V
8
11469



140
V W G P S G Y G G I
8
11470



154
C D R D D P S C D V
8
11471



174
L Q D L E D M S V M
8
11472



175
Q D L E D M S V M V
8
11473



183
M V L R T Q G P A A
8
11474



191
A A L F D D H K L V
8
11475



202
H T S S Y D A K R A
8
11476



204
S S Y D A K R A Q V
8
11477



252
E G L S F P D A G F
8
11478



260
G F T G L I S F H V
8
11479



276
N E D F S A S P I F
8
11480



277
E D F S A S P I F T
8
11481



284
I F T D T V V F R V
8
11482



290
V F R V A P W I M T
8
11483



293
V A P W I M T P S T
8
11484



311
C R V R N N T C F V
8
11485



315
N N T C F V D A V A
8
11486



343
K N D R W I Q D E M
8
11487



348
I Q D E M E L G Y V
8
11488



358
Q A P H K T L P V V
8
11489



392
V T R E P R D R S V
8
11490



398
D R S V S G L D S F
8
11491



407
F G N L E V S P P V
8
11492



408
G N L E V S P P V V
8
11493



433
N L P G S S G R R V
8
11494



440
R R V T Q V V R D F
8
11495



462
F V D W L A V G H V
8
11496



465
W L A V G H V D E F
8
11497



486
F R M L L A S P G A
8
11498



488
M L L A S P G A C F
8
11499



502
E K Q K C G H G R A
8
11500



509
G R A L L F Q G V V
8
11501



516
G V V D D E Q V K T
8
11502



517
V V D D E Q V K T I
8
11503



535
K D L I N Y N K F V
8
11504



553
E V L K R E L G L A
8
11505



558
E L G L A E C D I I
8
11506



566
I I D I P Q L F K T
8
11507



571
Q L F K T E R K K A
8
11508



579
K A T A F F P D L V
8
11509



581
T A F F P D L V N M
8
11510



590
H L V L G K H L G I
8
11511



598
G I P K P F G P I I
8
11512



639
H L H G E V H C G T
8
11513



646
C G T N V C R K P F
8
11514



654
P F S F K W W N M V
8
11515



19
C V A G V E T L V D
7
11516



24
E T L V D I Y G S V
7
11517



36
G T E M F E V Y G T
7
11518



43
Y G T P G V D I Y I
7
11519



100
H E P L P L A Y A V
7
11520



104
P L A Y A V L Y L T
7
11521



182
V M V L R T Q G P A
7
11522



185
L R T Q G P A A L F
7
11523



207
D A K R A Q V F H I
7
11524



220
E D V C E A Y R H V
7
11525



258
D A G F T G L I S F
7
11526



272
L D D S N E D F S A
7
11527



275
S N E D F S A S P I
7
11528



288
T V V F R V A P W I
7
11529



308
V Y V C R V R N N T
7
11530



321
D A V A E L A R K A
7
11531



331
G C K L T I C P Q A
7
11532



375
E L Q D F P Y K R I
7
11533



380
P Y K R I L G P D F
7
11534



446
V R D F L H A Q K V
7
11535



453
Q K V Q P P V E L F
7
11536



469
G H V D E F L S F V
7
11537



477
F V P A P D G K G F
7
11538



514
F Q G V V D D E Q V
7
11539



519
D D E Q V K T I S I
7
11540



534
N K D L I N Y N K F
7
11541



539
N Y N K F V Q S C I
7
11542



560
G L A E C D I I D I
7
11543



572
L F K T E R K K A T
7
11544



573
F K T E R K K A T A
7
11545



594
C K H L G I P K P F
7
11546



620
L L E P L G L H C T
7
11547



626
L H C T F I D D F T
7
11548



630
F I D D F T P Y H M
7
11549



641
H G E V H C G T N V
7
11550



68
W R F D A T L E I I
6
11551



83
P S N D L N D S H V
6
11552



162
D V Q D N C D Q H V
6
11553



171
V H C L Q D L E D M
6
11554



173
C L Q D L E D M S V
6
11555



186
R T Q G P A A L F D
6
11556



199
L V L H T S S Y D A
6
11557



208
A K R A Q V F H I C
6
11558



213
V F H I C G P E D V
6
11559



226
Y R H V L G Q D K V
6
11560



230
L G Q D K V S Y E V
6
11561



240
P R L H G D E E R F
6
11562



270
T L L D D S N E D F
6
11563



289
V V F R V A P W I M
6
11564



318
C F V D A V A E L A
6
11565



339
Q A E N R N D R W I
6
11566



354
L G Y V Q A P H K T
6
11567



411
E V S P P V V A N G
6
11568



418
A N G K E Y P L G R
6
11569



419
N G K E Y P L G R I
6
11570



458
P V E L F V D W L A
6
11571



479
P A P D G K G F R M
6
11572



522
Q V K T I S I N Q V
6
11573



529
N Q V L S N K D L I
6
11574



545
Q S C I D W N R E V
6
11575



564
C D I I D I P Q L F
6
11576



580
A T A F F P D L V N
6
11577



583
F F P D L V N M L V
6
11578



608
N G C C C L E E K V
6
11579



621
L E P L G L H C T F
6
11580



625
G L H C T F I D D F
6
11581



648
T N V C R K P F S F
6
11582



60
R E R A D T R R W R
5
11583



72
A T L E I I V V N N
5
11584



80
N M S P S N D L N D
5
11585



144
S G Y G G I L L V N
5
11586



145
C Y G G I L L V N C
5
11587



156
R D D P S C D V Q D
5
11588



206
Y D A K R A Q V F H
5
11589



257
P D A G F T G L I S
5
11590



264
L I S F H V T L L D
5
11591



292
R V A P W I M T P S
5
11592



296
W I M T P S T L P P
5
11593



319
F V D A V A E L A R
5
11594



322
A V A E L A R K A G
5
11595



328
R K A G C K L T I C
5
11596



397
R D R S V S G L D S
5
11597



435
P G S S G R R V T Q
5
11598



438
S G R R V T Q V V R
5
11599



439
G R R V T Q V V R D
5
11600



506
C G H G R A L L F Q
5
11601



511
A L L F Q G V V D D
5
11602



525
T I S I N Q V L S N
5
11603



595
K H L G I P K P F G
5
11604



606
I I N G C C C L E E
5
11605



26
L V D I Y G S V P E
4
11606



58
R G R E R A D T R R
4
11607



116
D I S L D C D L N C
4
11608



126
E G R Q D R N F V D
4
11609



133
F V D K R Q W V W G
4
11610



215
H I C G P E D V C E
4
11611



223
C E A Y R H V L G Q
4
11612



225
A Y R H V L G Q D K
4
11613



228
H V L G Q D K V S Y
4
11614



273
D D S N E D F S A S
4
11615



283
P I F T D T V V F R
4
11616



286
T D T V V F R V A P
4
11617



312
R V R N N T C F V D
4
11618



330
A G C K L T I C P Q
4
11619



352
M E L G Y V Q A P H
4
11620



360
P H K T L P V V F D
4
11621



400
S V S G L D S F G N
4
11622



405
D S F G N L E V S P
4
11623



410
L E V S P P V V A N
4
11624



427
R I L I G G N L P G
4
11625



428
I L I G G N L P G S
4
11626



483
G K G F R M L L A S
4
11627



510
R A L L F Q G V V D
4
11628



548
I D W N R E V L K R
4
11629



577
R K K A T A F F P D
4
11630



616
K V R S L L E P L G
4
11631



3
L Q R I V R V S L E
3
11632



21
A G V E T L V D I Y
3
11633



34
P E G T E M F E V Y
3
11634



35
E G T E M F E V Y G
3
11635



38
E M F E V Y G T P G
3
11636



51
Y I S P N M E R G R
3
11637



62
R A D T R R W R F D
3
11638



98
S S H E P L P L A Y
3
11639



105
L A Y A V L Y L T C
3
11640



127
G R Q D R N F V D K
3
11641



128
R Q D R N F V D K R
3
11642



135
D K R Q W V W G P S
3
11643



177
L E D M S V M V L R
3
11644



193
L F D D H K L V L H
3
11645



203
T S S Y D A K R A Q
3
11646



211
A Q V F H I C C P E
3
11647



222
V C E A Y R H V L G
3
11648



231
G Q D K V S Y E V P
3
11649



232
Q D K V S Y E V P R
3
11650



234
K V S Y E V P R L H
3
11651



235
V S Y E V P R L H G
3
11652



243
H G D E E R F F V E
3
11653



244
G D E E R F F V E G
3
11654



246
E E R F F V E G L S
3
11655



259
A G F T G L I S F H
3
11656



265
I S F H V T L L D D
3
11657



278
D F S A S P I F T D
3
11658



287
D T V V F R V A P W
3
11659



302
T L P P L E V Y V C
3
11660



306
L E V Y V C R V R N
3
11661



316
N T C F V D A V A E
3
11662



324
A E L A R K A G C K
3
11663



329
K A G C K L T I C P
3
11664



340
A E N R N D R W I Q
3
11665



341
E N R N D R W I Q D
3
11666



351
E M E L G Y V Q A P
3
11667



353
E L G Y V Q A P H K
3
11668



362
K T L P V V F D S P
3
11669



368
F D S P R N G E L Q
3
11670



371
P R N G E L Q D F P
3
11671



372
R N G E L Q D F P Y
3
11672



381
Y K R I L G P D F G
3
11673



387
P D F G Y V T R E P
3
11674



388
D F G Y V T R E P R
3
11675



393
T R E P R D R S V S
3
11676



394
R E P R D R S V S G
3
11677



404
L D S F G N L E V S
3
11678



406
S F G N L E V S P P
3
11679



417
V A N G K E Y P L G
3
11680



422
E Y P L G R I L I G
3
11681



429
L I G G N L P G S S
3
11682



445
V V R D F L H A Q K
3
11683



449
F L H A Q K V Q P P
3
11684



451
H A Q K V Q P P V E
3
11685



460
E L F V D W L A V G
3
11686



464
D W L A V G H V D E
3
11687



467
A V G H V D E F L S
3
11688



472
D E F L S F V P A P
3
11689



473
E F L S F V P A P D
3
11690



474
F L S F V P A P D G
3
11691



485
G F R M L L A S P G
3
11692



495
A C F K L F Q E K Q
3
11693



524
K T I S I N Q V L S
3
11694



537
L I N Y N K F V Q S
3
11695



547
C I D W N R E V L K
3
11696



554
V L K R E L G L A E
3
11697



555
L K R E L G L A E C
3
11698



562
A E C D I I D I P Q
3
11699



565
D I I D I P Q L F K
3
11700



567
I D I P Q L F K T E
3
11701



576
E R K K A T A F F P
3
11702



586
D L V N M L V L G K
3
11703



592
V L G K H L G I P K
3
11704



613
L E E K V R S L L E
3
11705



614
E E K V R S L L E P
3
11706



628
C T F I D D F T P Y
3
11707



633
D F T P Y H M L H G
3
11708



637
Y H M L H G E V H C
3
11709



643
E V H C G T N V C R
3
11710



644
V H C G T N V C R K
3
11711



650
V C R K P F S F K W
3
11712



1
M S L Q R I V R V S
2
11713



4
Q R I V R V S L E H
2
11714



10
S L E H P T S A V C
2
11715



15
T S A V C V A G V E
2
11716



27
V D I Y G S V P E G
2
11717



29
I Y G S V P E G T E
2
11718



42
V Y G T P G V D I Y
2
11719



44
G T P G V D I Y I S
2
11720



52
I S P N M E R G R E
2
11721



57
E R G R E R A D T R
2
11722



66
R R W R F D A T L E
2
11723



75
E I I V V M N S P S
2
11724



76
I I V V M N S P S N
2
11725



84
S N D L N D S H V Q
2
11726



88
N D S H V Q I S Y H
2
11727



93
Q I S Y H S S H E P
2
11728



102
P L P L A Y A V L Y
2
11729



107
Y A V L Y L T C V D
2
11730



112
L T C V D I S L D C
2
11731



129
Q D R N F V D K R Q
2
11732



136
K R Q W V W G P S G
2
11733



139
W V W G P S G Y G G
2
11734



146
Y G G I L L V N C D
2
11735



155
D R D D P S C D V Q
2
11736



163
V Q D N C D Q H V H
2
11737



164
Q D N C D Q H V H C
2
11738



167
C D Q H V H C L Q D
2
11739



170
H V H C L Q D L E D
2
11740



179
D H S V M V L R T Q
2
11741



180
M S V M V L R T Q G
2
11742



181
S V M V L R T Q G P
2
11743



187
T Q G P A A L F D D
2
11744



188
Q G P A A L F D D H
2
11745



209
K R A Q V F H I C G
2
11746



227
R H V L G Q D K V S
2
11747



229
V L G Q D K V S Y E
2
11748



248
R F F V E G L S F P
2
11749



251
V E G L S F P D A G
2
11750



271
L L D D S N E D F S
2
11751



307
E V Y V C R V R N N
2
11752



320
V D A V A E L A R K
2
11753



333
K L T I C P Q A E N
2
11754



344
N D R W I Q D E M E
2
11755



347
W I Q D E M E L G Y
2
11756



356
Y V Q A P H K T L P
2
11757



369
D S P R N G E L Q D
2
11758



373
N G E L Q D F P Y K
2
11759



377
Q D F P Y K R I L G
2
11760



378
D F P Y K R I L G P
2
11761



385
L G P D F G Y V T R
2
11762



399
R S V S G L D S F G
2
11763



430
I G G N L P G S S G
2
11764



447
R D F L H A Q K V Q
2
11765



448
D F L H A Q K V Q P
2
11766



455
V Q P P V E L F V D
2
11767



461
L F V D W L A V G H
2
11768



470
H V D E F L S F V P
2
11769



484
K G F R M L L A S P
2
11770



487
R M L L A S P G A C
2
11771



489
L L A S P G A C F K
2
11772



513
L F Q G V V D D E Q
2
11773



518
V D D E Q V K T I S
2
11774



521
E Q V K T I S I N Q
2
11775



530
Q V L S N K D L I N
2
11776



531
V L S N K D L I N Y
2
11777



538
I N Y N K F V Q S C
2
11778



544
V Q S C I D W N R E
2
11779



585
P D L V N M L V L G
2
11780



591
L V L G K H L G I P
2
11781



602
P F G P I I N G C C
2
11782



607
I N G C C C L E E K
2
11783



610
C C C L E E K V R S
2
11784



619
S L L E P L G L H C
2
11785



623
P L G L H C T F I D
2
11786



629
T F I D D F T P Y H
2
11787



642
G E V H C G T N V C
2
11788



647
G T N V C R K P F S
2
11789



649
N V C R K P F S F K
2
11790



651
C R K P F S F K W W
2
11791



655
F S F K W W N M V P
2
11792



5
R I V R V S L E H P
1
11793



7
V R V S L E H P T S
1
11794



25
T L V D I Y G S V P
1
11795



32
S V P E G T E M F E
1
11796



37
T E M F E V Y G T P
1
11797



40
F E V Y G T P G V D
1
11798



46
P G V D I Y I S P N
1
11799



50
I Y I S P N M E R G
1
11800



55
N M E R G R E R A D
1
11801



59
G R E R A D T R R W
1
11802



73
T L E I I V V M N S
1
11803



77
I V V M N S P S N D
1
11804



86
D L N D S H V Q I S
1
11805



87
L N D S H V Q I S Y
1
11806



95
S Y H S S H E P L P
1
11807



109
V L Y L T C V D I S
1
11808



117
I S L D C D L N C E
1
11809



118
S L D C D L N C E G
1
11810



120
D C D L N C E G R Q
1
11811



122
D L N C E G R Q D R
1
11812



132
N F V D K R Q W V W
1
11813



134
V D K R Q W V W G P
1
11814



137
R Q W V W G P S G Y
1
11815



149
I L L V N C D R D D
1
11816



150
L L V N C D R D D P
1
11817



151
L V N C D R D D P S
1
11818



152
V N C D R D D P S C
1
11819



153
N C D R D D P S C D
1
11820



157
D D P S C D V Q D N
1
11821



160
S C D V Q D N C D Q
1
11822



166
N C D Q H V H C L Q
1
11823



195
D D H K L V L H T S
1
11824



196
D H K L V L H T S S
1
11825



197
H K L V L H T S S Y
1
11826



198
K L V L H T S S Y D
1
11827



200
V L H T S S Y D A K
1
11828



201
L H T S S Y D A K R
1
11829



210
R A Q V F H I C G P
1
11830



212
Q V F H I C G P E D
1
11831



214
F H I C G P E D V C
1
11832



217
C G P E D V C E A Y
1
11833



219
P E D V C E A Y R H
1
11834



224
E A Y R H V L G Q D
1
11835



237
Y E V P R L H G D E
1
11836



238
E V P R L H G D E E
1
11837



249
F F V E G L S F P D
1
11838



254
L S F P D A G F T G
1
11839



274
D S N E D F S A S P
1
11840



291
F R V A P W I M T P
1
11841



295
P W I M T P S T L P
1
11842



298
M T P S T L P P L E
1
11843



300
P S T L P P L E V Y
1
11844



305
P L E V Y V C R V R
1
11845



323
V A E L A R K A G C
1
11846



332
C K L T I C P Q A E
1
11847



335
T I C P Q A E N R N
1
11848



336
I C P Q A E N R N D
1
11849



338
P Q A E N R N D R W
1
11850



346
R W I Q D E M E L G
1
11851



349
Q D E M E L G Y V Q
1
11852



361
H K T L P V V F D S
1
11853



363
T L P V V F D S P R
1
11854



366
V V F D S P R N G E
1
11855



382
K R I L G P D F G Y
1
11856



389
F G Y V T R E P R D
1
11857



391
Y V T R E P R D R S
1
11858



396
P R D R S V S G L D
1
11859



412
V S P P V V A N G K
1
11860



415
P V V A N G K E Y P
1
11861



424
P L G R I L I G G N
1
11862



432
G N L P G S S G R R
1
11863



444
Q V V R D F L H A Q
1
11864



463
V D W L A V G H V D
1
11865



476
S F V P A P D G K G
1
11866



493
P G A C F K L F Q E
1
11867



494
G A C F K L F Q E K
1
11868



498
K L F Q E K Q K C G
1
11869



500
F Q E K Q K C G H G
1
11870



507
G H G R A L L F Q G
1
11871



512
L L F Q G V V D D E
1
11872



515
Q G V V D D E Q V K
1
11873



526
I S I N Q V L S N K
1
11874



532
L S N K D L I N Y N
1
11875



536
D L I N Y N K F V Q
1
11876



542
K F V Q S C I D W N
1
11877



551
N R E V L K R E L G
1
11878



556
K R E L G L A E C D
1
11879



559
L G L A E C D I I D
1
11880



568
D I P Q L F K T E R
1
11881



593
L G K H L G I P K P
1
11882



596
H L G I P K P F G P
1
11883



609
G C C C L E E K V R
1
11884



618
R S L L E P L G L H
1
11885



624
L G L H C T F I D D
1
11886



636
P Y H M L H G E V H
1
11887



638
H M L H G E V H C G
1
11888



640
L H G E V H C G T N
1
11889



645
H C G T N V C R K P
1
11890



652
R K P F S F K W W N
1
11891







184P1E2 v.2: HLA Peptide


Scoring Results B 0702 10-mers SYFPEITHI












5
T P S T L A P L E V
20
11892



10
A P L E V Y V C R V
19
11893



3
I M T P S T L A P L
14
11894



7
S T L A P L H V Y V
9
11895



1
P W I M T P S T L A
7
11896



2
W I M T P S T L A P
5
11897



8
T L A P L E V Y V C
4
11898



9
L A P L E V Y V C R
2
11899



4
M T P S T L A P L E
1
11900



6
P S T L A P L E V Y
1
11901







184P1E2 v.3: HLA Peptide


Scoring Results B 0702 10-mers SYFPEITHI












10
V P D G K G F R M L
24
11902



8
V P V P D G K G F R
12
11903



1
V D E F L S F V P V
10
11904



7
F V P V P D G K G F
6
11905



9
P V P D G K G F R M
6
11906



3
E F L S F V P V P D
4
11907



4
F L S F V P V P D G
4
11908



2
D E F L S F V P V P
3
11909

















TABLE XLII







184P1E2: HLA Peptide Scoring


Results B08 10-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID















No Data

















TABLE XLIII







184P1E2: HLA Peptide Scoring


Results B1510 10-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID















No Data

















TABLE XLIV







184P1E2: HLA Peptide Scoring


Results B2705 10-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID















No Data

















TABLE XLV







184P1E2: HLA Peptide Scoring


Results B2709 10-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID















No Data






















TABLE XLVI










SEQ.



Pos
1 2 3 4 5 6 7 8 9 0
score
ID NO.
















184P1E2 v.1: HLA Peptide


Scoring Results B4402 10-mers SYFPEITHI












421
K E Y P L G R I L I
25
11910



34
P E G T E M F E V Y
23
11911



621
L E P L G L H C T F
23
11912



245
D E E R F F V E G L
22
11913



276
N E D F S A S P I F
22
11914



552
R E V L K R E L G L
22
11915



575
T E R K K A T A F F
21
11916



557
R E L G L A E C D I
20
11917



582
A F F P D L V N M L
20
11918



491
A S P G A C F K L F
19
11919



192
A L F D D H K L V L
18
11920



101
E P L P L A Y A V L
17
11921



282
S P I F T D T V V F
17
11922



480
A P D G K G F R M L
17
11923



546
S C I D W N R E V L
17
11924



563
E C D I I D I P Q L
17
11925



359
A P H K T L P V V F
16
11926



472
D E F L S F V P A P
16
11927



651
C R K P F S F K W W
16
11928



11
L E H P T S A V C V
15
11929



17
A V C V A G V E T L
15
11930



98
S S H E P L P L A Y
15
11931



100
H E P L P L A Y A V
15
11932



102
P L P L A Y A V L Y
15
11933



247
E R F F V E G L S F
15
11934



263
G L I S F H V T L L
15
11935



294
A P W I M T P S T L
15
11936



324
A E L A R K A G C K
15
11937



340
A E N R N D R W I Q
15
11938



382
K R I L G P D F G Y
15
11939



452
A Q K V Q P P V E L
15
11940



490
L A S P G A C F K L
15
11941



534
N K D L I N Y N K F
15
11942



562
A E C D I I D I P Q
15
11943



564
C D I I D I P Q L F
15
11944



574
K T E R K K A T A F
15
11945



597
L G I P K P F G P I
15
11946



2
S L Q R I V R V S L
14
11947



21
A G V E T L V D I Y
14
11948



23
V E T L V D I Y G S
14
11949



31
G S V P E G T E M F
14
11950



61
E R A D T R R W R F
14
11951



74
L E I I V V M N S P
14
11952



142
G P S G Y G G I L L
14
11953



184
V L R T Q G P A A L
14
11954



205
S Y D A K R A Q V F
14
11955



251
V E G L S F P D A G
14
11956



255
S F P D A G F T G L
14
11957



297
I M T P S T L P P L
14
11958



325
E L A R K A G C K L
14
11959



395
E P R D R S V S G L
14
11960



410
L E V S P P V V A N
14
11961



459
V E L F V D W L A V
14
11962



503
K Q K C G H G R A L
14
11963



505
K C G H G R A L L F
14
11964



517
V V D D E Q V K T I
14
11965



531
V L S N K D L I N Y
14
11966



541
N K F V Q S C I D W
14
11967



584
F P D L V N M L V L
14
11968



588
V N M L V L G K H L
14
11969



594
G K H L G I P K P F
14
11970



614
E E K V R S L L E P
14
11971



615
E K V R S L L E P L
14
11972



617
V R S L L E P L G L
14
11973



646
C G T N V C R K P F
14
11974



59
G R E R A D T R R W
13
11975



78
V V M N S P S N D L
13
11976



85
N D L N D S H V Q I
13
11977



96
Y H S S H E P L P L
13
11978



103
L P L A Y A V L Y L
13
11979



108
A V L Y L T C V D I
13
11980



110
L Y L T C V D I S L
13
11981



124
N C E G R Q D R N F
13
11982



125
C E G R Q D R N F V
13
11983



132
N F V D K R Q W V W
13
11984



165
D N C D Q H V H C L
13
11985



176
D L E D M S V M V L
13
11986



177
L E D M S V M V L R
13
11987



190
P A A L F D D H K L
13
11988



217
C G P E D V C E A Y
13
11989



221
D V C E A Y R H V L
13
11990



228
H V L G Q D K V S Y
13
11991



237
Y E V P R L H G D E
13
11992



246
E E R F F V E G L S
13
11993



252
E G L S F P D A G F
13
11994



262
T G L I S F H V T L
13
11995



300
P S T L P P L E V Y
13
11996



317
T C F V D A V A E L
13
11997



327
A R K A G C K L T I
13
11998



350
D E M E L G Y V Q A
13
11999



355
G Y V Q A P H K T L
13
12000



370
S P R N G E L Q D F
13
12001



374
G E L Q D F P Y K R
13
12002



375
E L Q D F P Y K R I
13
12003



394
R E P R D R S V S G
13
12004



440
R R V T Q V V R D F
13
12005



456
Q P P V E L F V D W
13
12006



477
F V P A P D G K G F
13
12007



504
Q K C G H G R A L L
13
12008



520
D E Q V K T I S I N
13
12009



578
K K A T A F F P D L
13
12010



604
G P I I N G C C C L
13
12011



611
C C L E E K V R S L
13
12012



625
G L H C T F I D D F
13
12013



628
C T F I D D F T P Y
13
12014



642
G E V H C G T N V C
13
12015



37
T E M F E V Y G T P
12
12016



40
F E V Y G T P G V D
12
12017



41
E V Y G T P G V D I
12
12018



42
V Y G T P G V D I Y
12
12019



43
Y G T P G V D I Y I
12
12020



56
M E R G R E R A D T
12
12021



60
R E R A D T R R W R
12
12022



68
W R F D A T L E I I
12
12023



87
L N D S H V Q I S Y
12
12024



130
D R N F V D K R Q W
12
12025



185
L R T Q G P A A L F
12
12026



197
H K L V L H T S S Y
12
12027



219
P E D V C E A Y R H
12
12028



223
C E A Y R H V L G Q
12
12029



241
R L H G D E E R F F
12
12030



258
D A G F T G L I S F
12
12031



270
T L L D D S N E D F
12
12032



306
L E V Y V C R V R N
12
12033



347
W I Q D E M E L G Y
12
12034



352
M E L G Y V Q A P H
12
12035



376
L Q D F P Y K R I L
12
12036



398
D R S V S G L D S F
12
12037



401
V S G L D S F G N L
12
12038



414
P P V V A N G K E Y
12
12039



453
Q K V Q P P V E L F
12
12040



468
V G H V D E F L S F
12
12041



481
P D G K G F R M L L
12
12042



528
I N Q V L S N K D L
12
12043



560
G L A E C D I I D I
12
12044



612
C L E E K V R S L L
12
12045



613
L E E K V R S L L E
12
12046



622
E P L G L H C T F I
12
12047



631
I D D F T P Y H M L
12
12048



65
T R R W R F D A T L
11
12049



114
C V D I S L D C D L
11
12050



137
R Q W V W G P S G Y
11
12051



140
V W G P S G Y G G I
11
12052



141
W G P S G Y G G I L
11
12053



168
D Q H V H C L Q D L
11
12054



233
D K V S Y E V P R L
11
12055



240
P R L H G D E E R F
11
12056



256
F P D A G F T G L I
11
12057



275
S N E D F S A S P I
11
12058



287
D T V V F R V A P W
11
12059



367
V F D S P R N G E L
11
12060



380
P Y K R I L G P D F
11
12061



416
V V A N G K E Y P L
11
12062



420
G K E Y P L G R I L
11
12063



441
R V T Q V V R D F L
11
12064



457
P P V E L F V D W L
11
12065



465
W L A V G H V D E F
11
12066



466
L A V G H V D E F L
11
12067



488
M L L A S P G A C F
11
12068



501
Q E K Q K C G H G R
11
12069



523
V K T I S I N Q V L
11
12070



529
N Q V L S N K D L I
11
12071



550
W N R E V L K R E L
11
12072



558
E L G L A E C D I I
11
12073



648
T N V C R K P F S F
11
12074



650
V C R K P F S F K W
11
12075



20
V A G V E T L V D I
10
12076



67
R W R F D A T L E I
10
12077



94
I S Y H S S H E P L
10
12078



207
D A K R A Q V F H I
10
12079



310
V C R V R N N T C F
10
12080



338
P Q A E N R N D R W
10
12081



339
Q A E N R N D R W I
10
12082



345
D R W I Q D E M E L
10
12083



372
R N G E L Q D F P Y
10
12084



419
N G K E Y P L G R I
10
12085



425
L G R I L I G G N L
10
12086



519
D D E Q V K T I S I
10
12087



567
I D I P Q L F K T E
10
12088



590
M L V L G K H L G I
10
12089



259
A G F T G L I S F H
9
12090



288
T V V F R V A P W I
9
12091



539
N Y N K F V Q S C I
9
12092



598
G I P K P F G P I I
9
12093



322
A V A E L A R K A G
8
12094



377
Q D F P Y K R I L G
8
12095



422
E Y P L G R I L I G
8
12096



601
K P F G P I I N G C
8
12097



191
A A L F D D H K L V
7
12098



280
S A S P I F T D T V
7
12099



411
E V S P P V V A N G
7
12100



12
E H P T S A V C V A
6
12101



70
F D A T L E I I V V
6
12102



71
D A T L E I I V V M
6
12103



72
A T L E I I V V M N
6
12104



156
R D D P S C D V Q D
6
12105



254
L S F P D A G F T G
6
12106



277
E D F S A S P I F T
6
12107



301
S T L P P L E V Y V
6
12108



302
T L P P L E V Y V C
6
12109



330
A G C K L T I C P Q
6
12110



351
E M E L G Y V Q A P
6
12111



362
K T L P V V F D S P
6
12112



412
V S P P V V A N G K
6
12113



426
G R I L I G G N L P
6
12114



447
R D F L H A Q K V Q
6
12115



455
V Q P P V E L F V D
6
12116



460
E L F V D W L A V G
6
12117



495
A C F K L F Q E K Q
6
12118



511
A L L F Q G V V D D
6
12119



522
Q V K T I S I N Q V
6
12120



524
K T I S I N Q V L S
6
12121



571
Q L F K T E R K K A
6
12122



44
G T P G V D I Y I S
5
12123



48
V D I Y I S P N M E
5
12124



51
Y I S P N M E R G R
5
12125



55
N M E R G R E R A D
5
12126



57
E R G R E R A D T R
5
12127



63
A D T R R W R F D A
5
12128



75
E I I V V M N S P S
5
12129



80
M N S P S N D L N D
5
12130



84
S N D L N D S H V Q
5
12131



88
N D S H V Q I S Y H
5
12132



99
S H E P L P L A Y A
5
12133



143
P S G Y G G I L L V
5
12134



144
S G Y G G I L L V N
5
12135



178
E D M S V M V L R T
5
12136



193
L F D D H K L V L H
5
12137



208
A K R A Q V F H I C
5
12138



224
E A Y R H V L G Q D
5
12139



234
K V S Y E V P R L H
5
12140



273
D D S N E D F S A S
5
12141



278
D F S A S P I F T D
5
12142



281
A S P I F T D T V V
5
12143



283
P I F T D T V V F R
5
12144



285
F T D T V V F R V A
5
12145



316
N T C F V D A V A E
5
12146



332
C K L T I C P Q A E
5
12147



341
E N R N D R W I Q D
5
12148



342
N R N D R W I Q D E
5
12149



357
V Q A P H K T L P V
5
12150



366
V V F D S P R N G E
5
12151



368
F D S P R N G E L Q
5
12152



378
D F P Y K R I L G P
5
12153



387
P D F G Y V T R E P
5
12154



393
T R E P R D R S V S
5
12155



402
S G L D S F G N L E
5
12156



418
A N G K F Y P L G R
5
12157



428
I L I G G N L P G S
5
12158



444
Q V V R D F L H A Q
5
12159



454
K V Q P P V F L F V
5
12160



476
S F V P A P D G K G
5
12161



484
K G F R M L L A S P
5
12162



487
R M L L A S P G A C
5
12163



498
K L F Q E K Q K C G
5
12164



526
I S I N Q V L S N K
5
12165



536
D L I N Y N K F V Q
5
12166



553
E V L K R E L G L A
5
12167



565
D I I D I P Q L F K
5
12168



572
L F K T E R K K A T
5
12169



580
A T A F F P D L V N
5
12170



599
I P K P F G P I I N
5
12171



600
P K P F G P I I N G
5
12172



605
P I I N G C C C L E
5
12173



620
L L E P L G L H C T
5
12174



629
T F I D D F T P Y H
5
12175



1
M S L Q R I V R V S
4
12176



4
Q R I V R V S L E H
4
12177



9
V S L E H P T S A V
4
12178



19
C V A G V E T L V D
4
12179



27
V D I Y G S V P E G
4
12180



38
E M F E V Y G T P G
4
12181



46
P G V D I Y I S P N
4
12182



64
D T R R W R F D A T
4
12183



81
N S P S N D L N D S
4
12184



106
A Y A V L Y L T C V
4
12185



111
Y L T C V D I S L D
4
12186



115
V D I S L D C D L N
4
12187



117
I S L D C D L N C E
4
12188



121
C D L N C E G R Q D
4
12189



128
R Q D R N F V D K R
4
12190



147
G G I L L V N C D R
4
12191



153
N C D R D D P S C D
4
12192



163
V Q D N C D Q H V H
4
12193



179
D M S V M V L R T Q
4
12194



183
M V L R T Q G P A A
4
12195



187
T Q G P A A L F D D
4
12196



202
H T S S Y D A K R A
4
12197



203
T S S Y D A K R A Q
4
12198



204
S S Y D A K R A Q V
4
12199



214
F H I C G P E D V C
4
12200



216
I C G P E D V C E A
4
12201



227
R H V L G Q D K V S
4
12202



238
E V P R L H G D E E
4
12203



261
F T G L I S F H V T
4
12204



264
L I S F H V T L L D
4
12205



265
I S F H V T L L D D
4
12206



286
T D T V V F R V A P
4
12207



289
V V F R V A P W I M
4
12208



291
F R V A P W I M T P
4
12209



295
P W I M T P S T L P
4
12210



296
W I M T P S T L P P
4
12211



299
T P S T L P P L E V
4
12212



307
E V Y V C R V R N N
4
12213



314
R N N T C F V D A V
4
12214



319
F V D A V A E L A R
4
12215



334
L T I C P Q A E N R
4
12216



335
T I C P Q A E N R N
4
12217



346
R W I Q D E M E L G
4
12218



369
D S P R N G E L Q D
4
12219



384
I L G P D F G Y V T
4
12220



385
L G P D F G Y V T R
4
12221



405
D S F G N L E V S P
4
12222



409
N L E V S P P V V A
4
12223



423
Y P L G R I L I G G
4
12224



424
P L G R I L I G G N
4
12225



432
G N L P G S S G R R
4
12226



433
N L P G S S G R R V
4
12227



435
P G S S G R R V T Q
4
12228



436
G S S G R R V T Q V
4
12229



437
S S G R R V T Q V V
4
12230



438
S G R R V T Q V V R
4
12231



467
A V G H V D E F L S
4
12232



473
E F L S F V P A P D
4
12233



483
G K G F R M L L A S
4
12234



496
C F K L F Q E K Q K
4
12235



510
R A L L F Q G V V D
4
12236



532
L S N K D L I N Y N
4
12237



535
K D L I N Y N K F V
4
12238



542
K F V Q S C I D W N
4
12239



548
I D W N R E V L K R
4
12240



549
D W N R E V L K R E
4
12241



554
V L K R E L G L A E
4
12242



581
T A F F P D L V N M
4
12243



585
P D L V N M L V L G
4
12244



589
N M L V L G K H L G
4
12245



593
L G K H L G I P K P
4
12246



618
R S L L E P L G L H
4
12247



632
D D F T P Y H M L H
4
12248



633
D F T P Y H M L H G
4
12249



8
R V S L E H P T S A
3
12250



13
H P T S A V C V A G
3
12251



24
E T L V D I Y G S V
3
12252



35
E G T E M F E V Y G
3
12253



50
I Y I S P N M E R G
3
12254



52
I S P N M E R G R E
3
12255



62
R A D T R R W R F D
3
12256



66
R R W R F D A T L E
3
12257



82
S P S N D L N D S H
3
12258



92
V Q I S Y H S S H E
3
12259



97
H S S H E P L P L A
3
12260



107
Y A V L Y L T C V D
3
12261



116
D I S L D C D L N C
3
12262



126
E G R Q D R N F V D
3
12263



127
G R Q D R N F V D K
3
12264



129
Q D R N F V D K R Q
3
12265



131
R N F V D K R Q W V
3
12266



145
G Y G G I L L V N C
3
12267



155
D R D D P S C D V Q
3
12268



166
N C D Q H V H C L Q
3
12269



174
L Q D L E D M S V M
3
12270



181
S V M V L R T Q G P
3
12271



186
R T Q G P A A L F D
3
12272



194
F D D H K L V L H T
3
12273



195
D D H K L V L H T S
3
12274



200
V L H T S S Y D A K
3
12275



210
R A Q V F H I C G P
3
12276



211
A Q V F H I C G P E
3
12277



220
E D V C E A Y R H V
3
12278



225
A Y R H V L G Q D K
3
12279



236
S Y E V P R L H G D
3
12280



243
H G D E E R F F V E
3
12281



248
R F F V E G L S F P
3
12282



269
V T L L D D S N E D
3
12283



292
R V A P W I M T P S
3
12284



298
M T P S T L P P L E
3
12285



303
L P P L E V Y V C R
3
12286



309
Y V C R V R N N T C
3
12287



313
V R N N T C F V D A
3
12288



318
C F V D A V A E L A
3
12289



321
D A V A E L A R K A
3
12290



326
L A R K A G C K L T
3
12291



329
K A G C K L T I C P
3
12292



353
E L G Y V Q A P H K
3
12293



354
L G Y V Q A P H K T
3
12294



358
Q A P H K T L P V V
3
12295



360
P H K T L P V V F D
3
12296



373
N G E L Q D F P Y K
3
12297



383
R I L G P D F G Y V
3
12298



390
G Y V T R E P R D R
3
12299



396
P R D R S V S G L D
3
12300



400
S V S G L D S F G N
3
12301



404
L D S F G N L E V S
3
12302



406
S F G N L E V S P P
3
12303



408
G N L E V S P P V V
3
12304



413
S P P V V A N G K E
3
12305



434
L P G S S G R R V T
3
12306



446
V R D F L H A Q K V
3
12307



448
D F L H A Q K V Q P
3
12308



461
L F V D W L A V G H
3
12309



463
V D W L A V G H V D
3
12310



475
L S F V P A P D G K
3
12311



478
V P A P D G K G F R
3
12312



482
D G K G F R M L L A
3
12313



506
C G H G R A L L F Q
3
12314



507
G H G R A L L F Q G
3
12315



512
L L F Q G V V D D E
3
12316



516
G V V D D E Q V K T
3
12317



521
E Q V K T I S I N Q
3
12318



525
T I S I N Q V L S N
3
12319



527
S I N Q V L S N K D
3
12320



537
L I N Y N K F V Q S
3
12321



547
C I D W N R E V L K
3
12322



551
N R E V L K R E L G
3
12323



556
K R E L G L A E C D
3
12324



561
L A E C D I I D I P
3
12325



566
I I D I P Q L F K T
3
12326



570
P Q L F K T E R K K
3
12327



576
E R K K A T A F F P
3
12328



579
K A T A F F P D L V
3
12329



583
F F P D L V N M L V
3
12330



586
D L V N M L V L G K
3
12331



591
L V L G K H L G I P
3
12332



595
K H L G I P K P F G
3
12333



602
P F G P I I N G C C
3
12334



609
G C C C L E E K V R
3
12335



619
S L L E P L G L H C
3
12336



624
L G L H C T F I D D
3
12337



630
F I D D F T P Y H M
3
12338



634
F T P Y H M L H G E
3
12339



637
Y H M L H G E V H C
3
12340



638
H M L H G E V H C G
3
12341



643
E V H C G T N V C R
3
12342



645
H C G T N V C R K P
3
12343



649
N V C R K P F S F K
3
12344



653
K P F S F K W W N M
3
12345



655
F S F K W W N M V P
3
12346



3
L Q R I V R V S L E
2
12347



5
R I V R V S L E H P
2
12348



10
S L E H P T S A V C
2
12349



14
P T S A V C V A G V
2
12350



15
T S A V C V A G V E
2
12351



16
S A V C V A G V E T
2
12352



22
G V E T L V D I Y G
2
12353



25
T L V D I Y G S V P
2
12354



26
L V D I Y G S V P E
2
12355



28
D I Y G S V P E G T
2
12356



32
S V P E G T E M F E
2
12357



53
S P N M E R G R E R
2
12358



54
P N M E R G R E R A
2
12359



69
R F D A T L E I I V
2
12360



86
D L N D S H V Q I S
2
12361



90
S H V Q I S Y H S S
2
12362



93
Q I S Y H S S H E P
2
12363



95
S Y H S S H E P L P
2
12364



104
P L A Y A V L Y L T
2
12365



105
L A Y A V L Y L T C
2
12366



109
V L Y L T C V D I S
2
12367



113
T C V D I S L D C D
2
12368



118
S L D C D L N C E G
2
12369



120
D C D L N C E G R Q
2
12370



133
F V D K R Q W V W G
2
12371



134
V D K R Q W V W G P
2
12372



146
Y G G I L L V N C D
2
12373



148
G I L L V N C D R D
2
12374



149
I L L V N C D R D D
2
12375



150
L L V N C D R D D P
2
12376



157
D D P S C D V Q D N
2
12377



159
P S C D V Q D N C D
2
12378



160
S C D V Q D N C D Q
2
12379



167
C D Q H V H C L Q D
2
12380



171
V H C L Q D L E D M
2
12381



175
Q D L E D M S V M V
2
12382



180
M S V M V L R T Q G
2
12383



188
Q G P A A L F D D H
2
12384



198
K L V L H T S S Y D
2
12385



209
K R A Q V F H I C G
2
12386



212
Q V F H I C G P E D
2
12387



215
H I C G P E D V C E
2
12388



222
V C E A Y R H V L G
2
12389



231
G Q D K V S Y E V P
2
12390



235
V S Y E V P R L H G
2
12391



242
L H G D E E R F F V
2
12392



244
G D E E R F F V E G
2
12393



250
F V E G L S F P D A
2
12394



253
G L S F P D A G F T
2
12395



257
P D A G F T G L I S
2
12396



266
S F H V T L L D D S
2
12397



271
L L D D S N E D F S
2
12398



284
I F T D T V V F R V
2
12399



293
V A P W I M T P S T
2
12400



304
P P L E V Y V C R V
2
12401



305
P L E V Y V C R V R
2
12402



308
V Y V C R V R N N T
2
12403



311
C R V R N N T C F V
2
12404



312
R V R N N T C F V D
2
12405



315
N N T C F V D A V A
2
12406



328
R K A G C K L T I C
2
12407



331
G C K L T I C P Q A
2
12408



336
I C P Q A E N R N D
2
12409



337
C P Q A E N R N D R
2
12410



343
R N D R W I Q D E M
2
12411



348
I Q D E M E L G Y V
2
12412



361
H K T L P V V F D S
2
12413



365
P V V F D S P R N G
2
12414



386
G P D F G Y V T R E
2
12415



391
Y V T R E P R D R S
2
12416



403
G L D S F G N L E V
2
12417



417
V A N G K E Y P L G
2
12418



427
R I L I G G N L P G
2
12419



429
L I G G N L P G S S
2
12420



439
G R R V T Q V V R D
2
12421



443
T Q V V R D F L H A
2
12422



445
V V R D F L H A Q K
2
12423



451
H A Q K V Q P P V E
2
12424



458
P V E L F V D W L A
2
12425



462
F V D W L A V G H V
2
12426



464
D W L A V G H V D E
2
12427



469
G H V D E F L S F V
2
12428



470
H V D E F L S F V P
2
12429



471
V D E F L S F V P A
2
12430



474
F L S F V P A P D G
2
12431



479
P A P D G K G F R M
2
12432



486
F R M L L A S P G A
2
12433



492
S P G A C F K L F Q
2
12434



493
P G A C F K L F Q E
2
12435



497
F K L F Q E K Q K C
2
12436



502
E K Q K C G H G R A
2
12437



509
G R A L L F Q G V V
2
12438



530
Q V L S N K D L I N
2
12439



533
S N K D L I N Y N K
2
12440



538
I N Y N K F V Q S C
2
12441



544
V Q S C I D W N R E
2
12442



559
L G L A E C D I I D
2
12443



569
I P Q L F K T E R K
2
12444



587
L V N M L V L G K H
2
12445



606
I I N G C C C L E E
2
12446



608
N G C C C L E E K V
2
12447



610
C C C L E E K V R S
2
12448



616
K V R S L L E P L G
2
12449



623
P L G L H C T F I D
2
12450



644
V H C G T N V C R K
2
12451



654
P F S F K W W N M V
2
12452



6
I V R V S L E H P T
1
12453



7
V R V S L E H P T S
1
12454



18
V C V A G V E T L V
1
12455



29
I Y G S V P H G T E
1
12456



30
Y G S V P E G T E M
1
12457



33
V P E G T E M F E V
1
12458



36
G T E M F E V Y G T
1
12459



39
M F E V Y G T P G V
1
12460



45
T P G V D I Y I S P
1
12461



47
G V D I Y I S P N M
1
12462



73
T L E I I V V M N S
1
12463



77
I V V M N S P S N D
1
12464



79
V M N S P S N D L N
1
12465



83
P S N D L N D S H V
1
12466



112
L T C V D I S L D C
1
12467



119
L D C D L N C E G R
1
12468



122
D L N C E G R Q D R
1
12469



136
K R Q W V W G P S G
1
12470



139
W V W G P S G Y G G
1
12471



152
V N C D R D D P S C
1
12472



154
C D R D D P S C D V
1
12473



158
D P S C D V Q D N C
1
12474



161
C D V Q D N C D Q H
1
12475



162
D V Q D N C D Q H V
1
12476



164
Q D N C D Q H V H C
1
12477



169
Q H V H C L Q D L E
1
12478



170
H V H C L Q D L E D
1
12479



172
H C L Q D L E D M S
1
12480



173
C L Q D L E D M S V
1
12481



182
V M V L R T Q G P A
1
12482



196
D H K L V L H T S S
1
12483



199
L V L H T S S Y D A
1
12484



201
L H T S S Y D A K R
1
12485



206
Y D A K R A Q V F H
1
12486



213
V F H I C G P E D V
1
12487



218
G P E D V C E A Y R
1
12488



226
Y R H V L G Q D K V
1
12489



229
V L G Q D K V S Y E
1
12490



230
L G Q D K V S Y E V
1
12491



232
Q D K V S Y E V P R
1
12492



239
V P R L H G D E E R
1
12493



260
G F T G L I S F H V
1
12494



267
F H V T L L D D S N
1
12495



268
H V T L L D D S N E
1
12496



272
L D D S N E D F S A
1
12497



279
F S A S P I F T D T
1
12498



290
V F R V A P W I M T
1
12499



320
V D A V A E L A R K
1
12500



323
V A E L A R K A G C
1
12501



333
K L T I C P Q A E N
1
12502



344
N D R W I Q D E M E
1
12503



349
Q D E M E L G Y V Q
1
12504



356
Y V Q A P H K T L P
1
12505



363
T L P V V F D S P R
1
12506



371
P R N G E L Q D F P
1
12507



379
F P Y K R I L G P D
1
12508



381
Y K R I L G P D F G
1
12509



388
D F G Y V T R E P R
1
12510



392
V T R E P R D R S V
1
12511



397
R D R S V S G L D S
1
12512



407
F G N L E V S P P V
1
12513



430
I G G N L P G S S G
1
12514



431
G G N L P G S S G R
1
12515



442
V T Q V V R D F L H
1
12516



449
F L H A Q K V Q P P
1
12517



485
G F R M L L A S P G
1
12518



489
L L A S P G A C F K
1
12519



494
G A C F K L F Q E K
1
12520



508
H G R A L L F Q G V
1
12521



513
L F Q G V V D D E Q
1
12522



514
F Q G V V D D E Q V
1
12523



515
Q G V V D D E Q V K
1
12524



518
V D D E Q V K T I S
1
12525



545
Q S C I D W N R E V
1
12526



555
L K R E L G L A E C
1
12527



568
D I P Q L F K T E R
1
12528



573
F K T E R K K A T A
I
12529



577
R K K A T A F F P D
1
12530



592
V L G K H L G I P K
1
12531



596
H L G I P K P F G P
1
12532



603
F G P I I N G C C C
1
12533



607
I N G C C C L E E K
1
12534



626
L H C T F I D D F T
1
12535



627
H C T F I D D F T P
1
12536



636
P Y H M L H G E V H
1
12537



639
M L H G E V H C G T
1
12538



640
L H G E V H C G T N
1
12539



652
R K P F S F K W W N
1
12540







184P1E2 v.2: HLA Peptide


Scoring Results B4402 10-mers SYFPEITHI












3
I M T P S T L A P L
14
12541



6
P S T L A P L E V Y
13
12542



2
W I M T P S T L A P
6
12543



8
T L A P L E V Y V C
5
12544



10
A P L E V Y V C R V
5
12545



1
P W I M T P S T L A
4
12546



5
T P S T L A P L E V
4
12547

















TABLE XLVII







184P1E2: HLA Peptide Scoring


Results B5101 10-mers SYFPEITHI















SEQ.



Pos
1 2 3 4 5 6 7 8 9
score
ID















No Data




















TABLE XLVIII








SEQ.


Pos
1 2 3 4 5 6 7 8 9 0 1 2 3 4 5
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results DRB1*0101 15-mers SYFPEITHI










258
D A G F T G L I S F H V T L L
33
12548


378
D F P Y K R I L G P D F G Y V
33
12549


179
D M S V M V L R T Q G P A A L
32
12550


23
V E T L V D I Y G S V P E G T
31
12551


182
V M V L R T Q G P A A L F D D
31
12552


423
Y P L G R I L I G G N L P G S
29
12553


483
G K G F R M L L A S P G A C F
29
12554


211
A Q V F H I C G P E D V C E A
28
12555


287
D T V V F R V A P W I M T P S
28
12556


37
T E M F E V Y G T P G V D I Y
27
12557


65
T R R W R F D A T L E I I V V
27
12558


104
P L A Y A V L Y L T C V D I S
27
12559


316
N T C F V D A V A E L A R K A
27
12560


404
L D S F G N L E V S P P V V A
27
12561


475
L S F V P A P D G K G F R M L
27
12562


594
G K H L G I P K P F G P I I N
27
12563


634
F T P Y H M L H G E V H C G T
27
12564


203
T S S Y D A K R A Q V F H I C
26
12565


223
C E A Y R H V L G Q D K V S Y
26
12566


286
T D T V V F R V A P W I M T P
26
12567


293
V A P W I M T P S T L P P L E
26
12568


444
Q V V R D F L H A Q K V Q P P
26
12569


581
T A F F P D L V N M L V L G K
26
12570


596
H L G I P K P F G P I I N G C
26
12571


615
E K V R S L L E P L G L H C T
26
12572


45
T P G V D I Y I S P N M E R G
25
12573


71
D A T L E I I V V M N S P S N
25
12574


171
V H C L Q D L E D M S V M V L
25
12575


323
V A E L A R K A G C K L T I C
25
12576


431
G G N L P G S S G R R V T Q V
25
12577


484
K G F R M L L A S P G A C F K
25
12578


6
I V R V S L E H P T S A V C V
24
12579


39
M F E V Y G T P G V D I Y I S
24
12580


106
A Y A V L Y L T C V D I S L D
24
12581


181
S V M V L R T Q G P A A L F D
24
12582


236
S Y E V P R L H G D E E R F F
24
12583


245
D E E R F F V E G L S F P D A
24
12584


248
R F F V E G L S F P D A G F T
24
12585


395
E P R D R S V S G L D S F G N
24
12586


398
D R S V S G L D S F G N L E V
24
12587


424
P L G R I L I G G N L P G S S
24
12588


460
E L F V D W L A V G H V D E F
24
12589


486
F R M L L A S P G A C F K L F
24
12590


586
D L V N M L V L G K H L G I P
24
12591


3
L Q R I V R V S L E H P T S A
23
12592


36
G T E M F E V Y G T P G V D I
23
12593


292
R V A P W I M T P S T L P P L
23
12594


439
G R R V T Q V V R D F L H A Q
23
12595


472
D E F L S F V P A P D G K G F
23
12596


520
D E Q V K T I S I N Q V L S N
23
12597


73
T L E I I V V M N S P S N D L
22
12598


76
I I V V M N S P S N D L N D S
22
12599


91
H V Q I S Y H S S H E P L P L
22
12600


174
L Q D L E D M S V M V L R T Q
22
12601


197
H K L V L H T S S Y D A K R A
22
12602


210
R A Q V F H I C G P E D V C E
22
12603


253
G L S F P D A G F T G L I S F
22
12604


294
A P W I M T P S T L P P L E V
22
12605


297
I M T P S T L P P L E V Y V C
22
12606


351
E M E L G Y V Q A P H K T L P
22
12607


407
F G N L E V S P P V V A N G K
22
12608


427
R I L I G G N L P G S S G R R
22
12609


447
R D F L H A Q K V Q P P V E L
22
12610


465
W L A V G H V D E F L S F V P
22
12611


468
V G H V D E F L S F V P A P D
22
12612


525
T I S I N Q V L S N K D L I N
22
12613


628
C T F I D D F T P Y H M L H G
22
12614


17
A V C V A G V E T L V D I Y G
21
12615


74
L E I I V V M N S P S N D L N
21
12616


138
Q W V W G P S G Y G G I L L V
21
12617


450
L H A Q K V Q P P V E L F V D
21
12618


459
V E L F V D W L A V G H V D E
21
12619


537
L I N Y N K F V Q S C I D W N
21
12620


569
I P Q L F K T E R K K A T A F
21
12621


87
L N D S H V Q I S Y H S S H E
20
12622


278
D F S A S P I F T D T V V F R
20
12623


353
E L G Y V Q A P H K T L P V V
20
12624


376
L Q D F P Y K R I L G P D F G
20
12625


379
F P Y K R I L G P D F G Y V T
20
12626


406
S F G N L E V S P P V V A N G
20
12627


506
C G H G R A L L F Q G V V D D
20
12628


517
V V D D E Q V K T I S I N Q V
20
12629


588
V N M L V L G K H L G I P K P
20
12630


112
L T C V D I S L D C D L N C E
19
12631


246
E E R F F V E G L S F P D A G
19
12632


282
S P I F T D T V V F R V A P W
19
12633


319
F V D A V A E L A R K A G C K
19
12634


388
D F G Y V T R E P R D R S V S
19
12635


551
N R E V L K R E L G L A E C D
19
12636


574
K T E R K K A T A F F P D L V
19
12637


585
P D L V N M L V L G K H L G I
19
12638


591
L V L G K H L G I P K P F G P
19
12639


5
R I V R V S L E H P T S A V C
18
12640


21
A G V E T L V D I Y G S V P E
18
12641


24
E T L V D I Y G S V P E G T E
18
12642


28
D I Y G S V P E G T E M F E V
18
12643


67
R W R F D A T L E I I V V M N
18
12644


93
Q I S Y H S S H E P L P L A Y
18
12645


108
A V L Y L T C V D I S L D C D
18
12646


137
R Q W V W G P S G Y G G I L L
18
12647


180
M S V M V L R T Q G P A A L F
18
12648


193
L F D D H K L V L H T S S Y D
18
12649


224
E A Y R H V L G Q D K V S Y E
18
12650


239
V P R L H G D E E R F F V E G
18
12651


264
L I S F H V T L L D D S N E D
18
12652


266
S F H V T L L D D S N E D F S
18
12653


365
P V V F D S P R N G E L Q D F
18
12654


413
S P P V V A N G K E Y P L G R
18
12655


426
G R I L I G G N L P G S S G R
18
12656


446
V R D F L H A Q K V Q P P V E
18
12657


501
Q E K Q K C G H G R A L L F Q
18
12658


556
K R E L G L A E C D I I D I P
18
12659


573
F K T E R K K A T A F F P D L
18
12660


580
A T A F F P D L V N M L V L G
18
12661


590
M L V L G K H L G I P K P F G
18
12662


614
E E K V R S L L E P L G L H C
18
12663


15
T S A V C V A G V E T L V D I
17
12664


68
W R F D A T L E I I V V M N S
17
12665


72
A T L E I I V V M N S P S N D
17
12666


81
N S P S N D L N D S H V Q I S
17
12667


84
S N D L N D S H V Q I S Y H S
17
12668


98
S S H E P L P L A Y A V L Y L
17
12669


100
H E P L P L A Y A V L Y L T C
17
12670


107
Y A V L Y L T C V D I S L D C
17
12671


132
N F V D K R Q W V W G P S G Y
17
12672


136
K R Q W V W G P S G Y G G I L
17
12673


140
V W G P S G Y G G I L L V N C
17
12674


148
G I L L V N C D R D D P S C D
17
12675


149
I L L V N C D R D D P S C D V
17
12676


183
M V L R T Q G P A A L F D D H
17
12677


194
F D D H K L V L H T S S Y D A
17
12678


200
V L H T S S Y D A K R A Q V F
17
12679


247
E R F F V E G L S F P D A G F
17
12680


251
V E G L S F P D A G F T G L I
17
12681


261
F T G L I S F H V T L L D D S
17
12682


290
V F R V A P W I M T P S T L P
17
12683


295
P W I M T P S T L P P L E V Y
17
12684


300
P S T L P P L E V Y V C R V R
17
12685


320
V D A V A E L A R K A G C K L
17
12686


343
R N D R W I Q D E M E L G Y V
17
12687


348
I Q D E M E L G Y V Q A P H K
17
12688


349
Q D E M E L G Y V Q A P H K T
17
12689


354
L G Y V Q A P H K T L P V V F
17
12690


363
T L P V V F D S P R N G E L Q
17
12691


373
N G E L Q D F P Y K R I L G P
17
12692


389
F G Y V T R E P R D R S V S G
17
12693


401
V S G L D S F G N L E V S P P
17
12694


418
A N G K E Y P L G R I L I G G
17
12695


419
N G K E Y P L G R I L I G G N
17
12696


455
V Q P P V E L F V D W L A V G
17
12697


456
Q P P V E L F V D W L A V G H
17
12698


462
F V D W L A V G H V D E F L S
17
12699


469
G H V D E F L S F V P A P D G
17
12700


487
R M L L A S P G A C F K L F Q
17
12701


507
G H G R A L L F Q G V V D D E
17
12702


515
Q G V V D D E Q V K T I S I N
17
12703


526
I S I N Q V L S N K D L I N Y
17
12704


535
K D L I N Y N K F V Q S C I D
17
12705


548
I D W N R E V L K R E L G L A
17
12706


552
R E V L K R E L G L A E C D I
17
12707


561
L A E C D I I D I P Q L F K T
17
12708


562
A E C D I I D I P Q L F K T E
17
12709


566
I I D I P Q L F K T E R K K A
17
12710


572
L F K T E R K K A T A F F P D
17
12711


584
F P D L V N M L V L G K H L G
17
12712


587
L V N M L V L G K H L G I P K
17
12713


601
K P F G P I I N G C C C L E E
17
12714


618
R S L L E P L G L H C T F I D
17
12715


620
L L E P L G L H C T F I D D F
17
12716


639
M L H G E V H C G T N V C R K
17
12717


9
V S L E H P T S A V C V A G V
16
12718


12
E H P T S A V C V A G V E T L
16
12719


20
V A G V E T L V D I Y G S V P
16
12720


26
L V D I Y G S V P E G T E M F
16
12721


27
V D I Y G S V P E G T E M F E
16
12722


29
I Y G S V P E G T E M F E V Y
16
12723


75
E I I V V M N S P S N D L N D
16
12724


97
H S S H E P L P L A Y A V L Y
16
12725


99
S H E P L P L A Y A V L Y L T
16
12726


139
W V W G P S G Y G G I L L V N
16
12727


143
P S G Y G G I L L V N C D R D
16
12728


152
V N C D R D D P S C D V Q D N
16
12729


160
S C D V Q D N C D Q H V H C L
16
12730


168
D Q H V H C L Q D L E D M S V
16
12731


190
P A A L F D D H K L V L H T S
16
12732


196
D H K L V L H T S S Y D A K R
16
12733


219
P E D V C E A Y R H V L G Q D
16
12734


229
V L G Q D K V S Y E V P R L H
16
12735


255
S F P D A G F T G L I S F H V
16
12736


268
H V T L L D D S N E D F S A S
16
12737


269
V T L L D D S N E D F S A S P
16
12738


273
D D S N E D F S A S P I F T D
16
12739


357
V Q A P H K T L P V V F D S P
16
12740


381
Y K R I L G P D F G Y V T R E
16
12741


382
K R I L G P D F G Y V T R E P
16
12742


386
G P D F G Y V T R E P R D R S
16
12743


405
D S F G N L E V S P P V V A N
16
12744


410
L E V S P P V V A N G K E Y P
16
12745


414
P P V V A N G K E Y P L G R I
16
12746


422
E Y P L G R I L I G G N L P G
16
12747


428
I L I G G N L P G S S G R R V
16
12748


438
S G R R V T Q V V R D F L H A
16
12749


452
A Q K V Q P P V E L F V D W L
16
12750


463
V D W L A V G H V D E F L S F
16
12751


471
V D E F L S F V P A P D G K G
16
12752


477
F V P A P D G K G F R M L L A
16
12753


503
K Q K C G H G R A L L F Q G V
16
12754


509
G R A L L F Q G V V D D E Q V
16
12755


511
A L L F Q G V V D D E Q V K T
16
12756


532
L S N K D L I N Y N K F V Q S
16
12757


555
L K R E L G L A E C D I I D I
16
12758


558
E L G L A E C D I I D I P Q L
16
12759


589
N M L V L G K H L G I P K P F
16
12760


592
V L G K H L G I P K P F G P I
16
12761


600
P K P F G P I I N G C C C L E
16
12762


610
C C C L E E K V R S L L E P L
16
12763


617
V R S L L E P L G L H C T F I
16
12764


623
P L G L H C T F I D D F T P Y
16
12765


625
G L H C T F I D D F T P Y H M
16
12766


627
H C T F I D D F T P Y H M L H
16
12767


644
V H C G T N V C R K P F S F K
16
12768


14
P T S A V C V A G V E T L V D
15
12769


88
N D S H V Q I S Y H S S H E P
15
12770


94
I S Y H S S H E P L P L A Y A
15
12771


109
V L Y L T C V D I S L D C D L
15
12772


146
Y G G I L L V N C D R D D P S
15
12773


169
Q H V H C L Q D L E D M S V M
15
12774


177
L E D M S V M V L R T Q G P A
15
12775


232
Q D K V S Y E V P R L H G D E
15
12776


262
T G L I S F H V T L L D D S N
15
12777


272
L D D S N E D F S A S P I F T
15
12778


274
D S N E D F S A S P I F T D T
15
12779


280
S A S P I F T D T V V F R V A
15
12780


283
P I F T D T V V F R V A P W I
15
12781


284
I F T D T V V F R V A P W I M
15
12782


309
Y V C R V R N N T C F V D A V
15
12783


314
R N N T C F V D A V A E L A R
15
12784


346
R W I Q D E M E L G Y V Q A P
15
12785


385
L G P D F G Y V T R E P R D R
15
12786


457
P P V E L F V D W L A V G H V
15
12787


485
G F R M L L A S P G A C F K L
15
12788


516
G V V D D E Q V K T I S I N Q
15
12789


577
R K K A T A F F P D L V N M L
15
12790


582
A F F P D L V N M L V L G K H
15
12791


621
L E P L G L H C T F I D D F T
15
12792


646
C G T N V C R K P F S F K W W
15
12793


7
V R V S L E H P T S A V C V A
14
12794


8
R V S L E H P T S A V C V A G
14
12795


42
V Y G T P G V D I Y I S P N M
14
12796


123
L N C E G R Q D R N F V D K R
14
12797


134
V D K R Q W V W G P S G Y G G
14
12798


163
V Q D N C D Q H V H C L Q D L
14
12799


173
C L Q D L E D M S V M V L R T
14
12800


195
D D H K L V L H T S S Y D A K
14
12801


226
Y R H V L G Q D K V S Y E V P
14
12802


228
H V L G Q D K V S Y E V P R L
14
12803


254
L S F P D A G F T G L I S F H
14
12804


302
T L P P L E V Y V C R V R N N
14
12805


307
E V Y V C R V R N N T C F V D
14
12806


317
T C F V D A V A E L A R K A G
14
12807


327
A R K A G C K L T I C P Q A E
14
12808


333
K L T I C P Q A E N R N D R W
14
12809


355
G Y V Q A P H K T L P V V F D
14
12810


360
P H K T L P V V F D S P R N G
14
12811


361
H K T L P V V F D S P R N G E
14
12812


435
P G S S G R R V T Q V V R D F
14
12813


449
F L H A Q K V Q P P V E L F V
14
12814


482
D G K G F R M L L A S P G A C
14
12815


512
L L F Q G V V D D E Q V K T I
14
12816


522
Q V K T I S I N Q V L S N K D
14
12817


538
I N Y N K F V Q S C I D W N R
14
12818


563
E C D I I D I P Q L F K T E R
14
12819


611
C C L E E K V R S L L E P L G
14
12820


633
D F T P Y H M L H G E V H C G
14
12821


352
M E L G Y V Q A P H K T L P V
13
12822


494
G A C F K L F Q E K Q K C G H
13
12823


46
P G V D I Y I S P N M F R G R
12
12824


234
K V S Y E V P R L H G D E E R
12
12825


279
F S A S P I F T D T V V F R V
12
12826


288
T V V F R V A P W I M T P S T
12
12827


291
F R V A P W I M T P S T L P P
12
12828


330
A G C K L T I C P Q A E N R N
12
12829


331
G C K L T I C P Q A E N R N D
12
12830


393
T R E P R D R S V S G L D S F
12
12831


425
L G R I L I G G N L P G S S G
12
12832


474
F L S F V P A P D G K G F R M
12
12833


497
F K L F Q E K Q K C G H G R A
12
12834


521
E Q V K T I S I N Q V L S N K
12
12835


533
S N K D L I N Y N K F V Q S C
12
12836


543
F V Q S C I D W N R E V L K R
12
12837


547
C I D W N R E V L K R E L G L
12
12838


550
W N R E V L K R E L G L A E C
12
12839


564
C D I I D I P Q L F K T E R K
12
12840


570
P Q L F K T E R K K A T A F F
12
12841


602
P F G P I I N G C C C L E E K
12
12842


1
M S L Q R I V R V S L E H P T
11
12843


40
F E V Y G T P G V D I Y I S P
11
12844


47
G V D I Y I S P N M E R G R E
11
12845


49
D I Y I S P N M E R G R E R A
11
12846


51
Y I S P N M E R G R E R A D T
11
12847


89
D S H V Q I S Y H S S H E P L
11
12848


130
D R N F V D K R Q W V W G P S
11
12849


144
S G Y G G I L L V N C D R D D
11
12850


225
A Y R H V L G Q D K V S Y E V
11
12851


244
G D E E R F F V E G L S F P D
11
12852


260
G F T G L I S F H V T L L D D
11
12853


306
L E V Y V C R V R N N T C F V
11
12854


308
V Y V C R V R N N T C F V D A
11
12855


344
N D R W I Q D E M E L G Y V Q
11
12856


368
F D S P R N G E L Q D F P Y K
11
12857


390
G Y V T R E P R D R S V S G L
11
12858


420
G K E Y P L G R I L I G G N L
11
12859


429
L I G G N L P G S S G R R V T
11
12860


493
P G A C F K L F Q E K Q K C G
11
12861


499
L F Q E K Q K C G H G R A L L
11
12862


500
F Q E K Q K C G H G R A L L F
11
12863


514
F Q G V V D D E Q V K T I S I
11
12864


523
V K T I S I N Q V L S N K D L
11
12865


540
Y N K F V Q S C I D W N R E V
11
12866


553
E V L K R E L G L A E C D I I
11
12867


631
I D D F T P Y H M L H G E V H
11
12868


648
T N V C R K P F S F K W W N M
11
12869


4
Q R I V R V S L E H P T S A V
10
12870


16
S A V C V A G V E T L V D I Y
10
12871


48
V D I Y I S P N M E R G R E R
10
12872


63
A D T R R W R F D A T L E I I
10
12873


69
R F D A T L E I I V V M N S P
10
12874


117
I S L D C D L N C E G R Q D R
10
12875


119
L D C D L N C E G R Q D R N F
10
12876


120
D C D L N C E G R Q D R N F V
10
12877


131
R N F V D K R Q W V W G P S G
10
12878


133
F V D K R Q W V W G P S G Y G
10
12879


188
Q G P A A L F D D H K L V L H
10
12880


189
G P A A L F D D H K L V L H T
10
12881


191
A A L F D D H K L V L H T S S
10
12882


208
A K R A Q V F H I C G P E D V
10
12883


218
G P E D V C E A Y R H V L G Q
10
12884


231
G Q D K V S Y E V P R L H G D
10
12885


243
H G D E E R F F V E G L S F P
10
12886


250
F V E G L S F P D A G F T G L
10
12887


256
F P D A G F T G L I S F H V T
10
12888


276
N E D F S A S P I F T D T V V
10
12889


310
V C R V R N N T C F V D A V A
10
12890


328
R K A G C K L T I C P Q A E N
10
12891


329
K A G C K L T I C P Q A E N R
10
12892


345
D R W I Q D E M E L G Y V Q A
10
12893


359
A P H K T L P V V F D S P R N
10
12894


364
L P V V F D S P R N G E L Q D
10
12895


374
G E L Q D F P Y K R I L G P D
10
12896


412
V S P P V V A N G K E Y P L G
10
12897


416
V V A N G K E Y P L G R I L I
10
12898


443
T Q V V R D F L H A Q K V Q P
10
12899


478
V P A P D G K G F R M L L A S
10
12900


479
P A P D G K G F R M L L A S P
10
12901


527
S I N Q V L S N K D L I N Y N
10
12902


607
I N G C C C L E E K V R S L L
10
12903


608
N G C C C L E E K V R S L L E
10
12904


637
Y H M L H G E V H C G T N V C
10
12905


643
E V H C G T N V C R K P F S F
10
12906


31
G S V P E G T E M F E V Y G T
9
12907


35
E G T E M F E V Y G T P G V D
9
12908


41
E V Y G T P G V D I Y I S P N
9
12909


52
I S P N M E R G R E R A D T R
9
12910


53
S P N M E R G R E R A D T R R
9
12911


61
E R A D T R R W R F D A T L E
9
12912


64
D T R R W R F D A T L E I I V
9
12913


77
I V V M N S P S N D L N D S H
9
12914


114
C V D I S L D C D L N C E G R
9
12915


122
D L N C E G R Q D R N F V D K
9
12916


128
R Q D R N F V D K R Q W V W G
9
12917


135
D K R Q W V W G P S G Y G G I
9
12918


141
W G P S G Y G G I L L V N C D
9
12919


145
G Y G G I L L V N C D R D D P
9
12920


147
G G I L L V N C D R D D P S C
9
12921


172
H C L Q D L E D M S V M V L R
9
12922


178
E D M S V M V L R T Q G P A A
9
12923


205
S Y D A K R A Q V F H I C G P
9
12924


213
V F H I C G P E D V C E A Y R
9
12925


217
C G P E D V C E A Y R H V L G
9
12926


240
P R L H G D E E R F F V E G L
9
12927


265
I S F H V T L L D D S N E D F
9
12928


270
T L L D D S N E D F S A S P I
9
12929


289
V V F R V A P W I M T P S T L
9
12930


303
L P P L E V Y V C R V R N N T
9
12931


305
P L E V Y V C R V R N N T C F
9
12932


311
C R V R N N T C F V D A V A E
9
12933


312
R V R N N T C F V D A V A E L
9
12934


313
V R N N T C F V D A V A E L A
9
12935


325
E L A R K A G C K L T I C P Q
9
12936


337
C P Q A E N R N D R W I Q D E
9
12937


347
W I Q D E M E L G Y V Q A P H
9
12938


358
Q A P H K T L P V V F D S P R
9
12939


369
D S P R N G E L Q D F P Y K R
9
12940


396
P R D R S V S G L D S F G N L
9
12941


399
R S V S G L D S F G N L E V S
9
12942


402
S G L D S F G N L E V S P P V
9
12943


417
V A N G K E Y P L G R I L I G
9
12944


434
L P G S S G R R V T Q V V R D
9
12945


441
R V T Q V V R D F L H A Q K V
9
12946


442
V T Q V V R D F L H A Q K V Q
9
12947


451
H A Q K V Q P P V E L F V D W
9
12948


461
L F V D W L A V G H V D E F L
9
12949


464
D W L A V G H V D E F L S F V
9
12950


466
L A V G H V D E F L S F V P A
9
12951


467
A V G H V D E F L S F V P A P
9
12952


470
H V D E F L S F V P A P D G K
9
12953


480
A P D G K G F R M L L A S P G
9
12954


481
P D G K G F R M L L A S P G A
9
12955


488
M L L A S P G A C F K L F Q E
9
12956


489
L L A S P G A C F K L F Q E K
9
12957


495
A C F K L F Q E K Q K C G H G
9
12958


496
C F K L F Q E K Q K C G H G R
9
12959


502
E K Q K C G H G R A L L F Q G
9
12960


508
H G R A L L F Q G V V D D E Q
9
12961


519
D D E Q V K T I S I N Q V L S
9
12962


531
V L S N K D L I N Y N K F V Q
9
12963


554
V L K R E L G L A E C D I I D
9
12964


571
Q L F K T E R K K A T A F F P
9
12965


579
K A T A F F P D L V N M L V L
9
12966


604
G P I I N G C C C L E E K V R
9
12967


606
I I N G C C C L E E K V R S L
9
12968


609
G C C C L E E K V R S L L E P
9
12969


612
C L E E K V R S L L E P L G L
9
12970


613
L E E K V R S L L E P L G L H
9
12971


619
S L L E P L G L H C T F I D D
9
12972


629
T F I D D F T P Y H M L H G E
9
12973


636
P Y H M L H G E V H C G T N V
9
12974


645
H C G T N V C R K P F S F K W
9
12975


10
S L E H P T S A V C V A G V E
8
12976


13
H P T S A V C V A G V E T L V
8
12977


18
V C V A G V E T L V D I Y G S
8
12978


22
G V E T L V D I Y G S V P E G
8
12979


30
Y G S V P E G T E M F E V Y G
8
12980


43
Y G T P C V D I Y I S P N M e
8
12981


55
N M E R G R E R A D T R R W R
8
12982


59
G R E R A D T R R W R F D A T
8
12983


66
R R W R F D A T L E I I V V M
8
12984


79
V M N S P S N D L N D S H V Q
8
12985


83
P S N D L N D S H V Q I S Y H
8
12986


92
V Q I S Y H S S H E P L P L A
8
12987


95
S Y H S S H E P L P L A Y A V
8
12988


101
E P L P L A Y A V L Y L T C V
8
12989


102
P L P L A Y A V L Y L T C V D
8
12990


105
L A Y A V L Y L T C V D I S L
8
12991


113
T C V D I S L D C D L N C E G
8
12992


116
D I S L D C D L N C E G R Q D
8
12993


129
Q D R N F V D K R Q W V W G P
8
12994


166
N C D Q H V H C L Q D L E D M
8
12995


170
H V H C L Q D L E D M S V M V
8
12996


184
V L R T Q G P A A L F D D H K
8
12997


187
T Q G P A A L F D D H K L V L
8
12998


198
K L V L H T S S Y D A K R A Q
8
12999


202
H T S S Y D A K R A Q V F H I
8
13000


206
Y D A K R A Q V F H I C G P E
8
13001


214
F H I C G P E D V C E A Y R H
8
13002


227
R H V L G Q D K V S Y E V P R
8
13003


238
E V P R L H G D E E R F F V E
8
13004


252
E G L S F P D A G F T G L I S
8
13005


271
L L D D S N E D F S A S P I F
8
13006


281
A S P I F T D T V V F R V A P
8
13007


299
T P S T L P P L E V Y V C R V
8
13008


315
N N T C F V D A V A E L A R K
8
13009


322
A V A E L A R K A G C K L T I
8
13010


332
C K L T I C P Q A E N R N D R
8
13011


341
E N R N D R W I Q D E M E L G
8
13012


342
N R N D R W I Q D E M E L G Y
8
13013


356
Y V Q A P H K T L P V V F D S
8
13014


366
V V F D S P R N G E L Q D F P
8
13015


370
S P R N G E L Q D F P Y K R I
8
13016


391
Y V T R E P R D R S V S G L D
8
13017


400
S V S G L D S F G N L E V S P
8
13018


403
G L D S F G N L E V S P P V V
8
13019


409
N L E V S P P V V A N G K E Y
8
13020


436
G S S G R R V T Q V V R D F L
8
13021


448
D F L H A Q K V Q P P V E L F
8
13022


458
P V E L F V D W L A V G H V D
8
13023


473
E F L S F V P A P D G K G F R
8
13024


498
K L F Q E K Q K C G H G R A L
8
13025


510
R A L L F Q G V V D D E Q V K
8
13026


528
I N Q V L S N K D L I N Y N K
8
13027


529
N Q V L S N K D L I N Y N K F
8
13028


534
N K D L I N Y N K F V Q S C I
8
13029


541
N K F V Q S C I D W N R E V L
8
13030


544
V Q S C I D W N R E V L K R E
8
13031


545
Q S C I D W N R E V L K R E L
8
13032


549
D W N R E V L K R E L G L A E
8
13033


559
L G L A E C D I I D I P Q L F
8
13034


576
E R K K A T A F F P D L V N M
8
13035


595
K H L G I P K P F G P I I N G
8
13036


597
L G I P K P F G P I I N G C C
8
13037


603
F G P I I N G C C C L E E K V
8
13038


640
L H G E V H C G T N V C R K P
8
13039


641
H G E V H C G T N V C R K P F
8
13040


647
G T N V C R K P F S F K W W N
8
13041


33
V P E G T E M F E V Y G T P G
7
13042


38
E M F E V Y G T P G V D I Y I
7
13043


44
G T P G V D I Y I S P N M E R
7
13044


50
I Y I S P N M E R G R E R A D
7
13045


57
E R G R E R A D T R R W R F D
7
13046


82
S P S N D L N D S H V Q I S Y
7
13047


111
Y L T C V D I S L D C D L N C
7
13048


249
F F V E G L S F P D A G F T G
7
13049


259
A G F T G L I S F H V T L L D
7
13050


267
F H V T L L D D S N E D F S A
7
13051


304
P P L E V Y V C R V R N N T C
7
13052


362
K T L P V V F D S P R N G E L
7
13053


371
P R N G E L Q D F P Y K R I L
7
13054


372
R N G E L Q D F P Y K R I L G
7
13055


411
E V S P P V V A N G K E Y P L
7
13056


453
Q K V Q P P V E L F V D W L A
7
13057


565
D I I D I P Q L F K T E R K K
7
13058


567
I D I P Q L F K T E R K K A T
7
13059


2
S L Q R I V R V S L E H P T S
6
13060


70
F D A T L E I I V V M N S P S
6
13061


86
D L N D S H V Q I S Y H S S H
6
13062


90
S H V Q I S Y H S S H E P L P
6
13063


96
Y H S S H E P L P L A Y A V L
6
13064


103
L P L A Y A V L Y L T C V D I
6
13065


110
L Y L T C V D I S L D C D L N
6
13066


151
L V N C D R D D P S C D V Q D
6
13067


157
D D P S C D V Q D N C D Q H V
6
13068


158
D P S C D V Q D N C D Q H V H
6
13069


165
D N C D Q H V H C L Q D L E D
6
13070


176
D L E D M S V M V L R T Q G P
6
13071


207
D A K R A Q V F H I C G P E D
6
13072


216
I C G P E D V C E A Y R H V L
6
13073


233
D K V S Y E V P R L H G D E E
6
13074


263
G L I S F H V T L L D D S N E
6
13075


275
S N E D F S A S P I F T D T V
6
13076


296
W I M T P S T L P P L E V Y V
6
13077


324
A E L A R K A G C K L T I C P
6
13078


392
V T R E P R D R S V S G L D S
6
13079


394
R E P R D R S V S G L D S F G
6
13080


430
I G G N L P G S S G R R V T Q
6
13081


440
R R V T Q V V R D F L H A Q K
6
13082


524
K T I S I N Q V L S N K D L I
6
13083


539
N Y N K F V Q S C I D W N R E
6
13084


542
K F V Q S C I D W N R E V L K
6
13085


560
G L A E C D I I D I P Q L F K
6
13086


593
L G K H L G I P K P F G P I I
6
13087


638
H M L H G E V H C G T N V C R
6
13088


34
P E G T E M F E V Y G T P G V
4
13089


237
Y E V P R L H G D E E R F F V
4
13090


626
L H C T F I D D F T P Y H M L
4
13091


54
P N M E R G R E R A D T R R W
3
13092


56
M E R G R E R A D T R R W R F
3
13093


58
R G R E R A D T R R W R F D A
3
13094


127
G R Q D R N F V D K R Q W V W
3
13095


321
D A V A E L A R K A G C K L T
3
13096


380
P Y K R I L G P D F G Y V T R
3
13097


530
Q V L S N K D L I N Y N K F V
3
13098


568
D I P Q L F K T E R K K A T A
3
13099


62
R A D T R R W R F D A T L E I
2
13100


124
N C E G R Q D R N F V D K R Q
2
13101


199
L V L H T S S Y D A K R A Q V
2
13102


204
S S Y D A K R A Q V F H I C G
2
13103


209
K R A Q V F H I C G P E D V C
2
13104


221
D V C E A Y R H V L G Q D K V
2
13105


298
M T P S T L P P L E V Y V C R
2
13106


334
L T I C P Q A E N R N D R W I
2
13107


339
Q A E N R N D R W I Q D E M E
2
13108


377
Q D F P Y K R I L G P D F G Y
2
13109


384
I L G P D F G Y V T R E P R D
2
13110


415
P V V A N G K E Y P L G R I L
2
13111


421
K E Y P L G R I L I G G N L P
2
13112


432
G N L P G S S G R R V T Q V V
2
13113


437
S S G R R V T Q V V R D F L H
2
13114


445
V V R D F L H A Q K V Q P P V
2
13115


546
S C I D W N R E V L K R E L G
2
13116


599
I P K P F G P I I N G C C C L
2
13117


632
D D F T P Y H M L H G E V H C
2
13118


635
T P Y H M L H G E V H C G T N
2
13119


649
N V C R K P F S F K W W N M V
2
13120


11
L E H P T S A V C V A G V E T
1
13121


19
C V A G V E T L V D I Y G S V
1
13122


60
R E R A D T R R W R F D A T L
1
13123


118
S L D C D L N C E G R Q D R N
1
13124


126
E G R Q D R N F V D K R Q W V
1
13125


159
P S C D V Q D N C D Q H V H C
1
13126


162
D V Q D N C D Q H V H C L Q D
1
13127


167
C D Q H V H C L Q D L E D M S
1
13128


175
Q D L E D M S V M V L R T Q G
1
13129


201
L H T S S Y D A K R A Q V F H
1
13130


230
L G Q D K V S Y E V P R L H G
1
13131


235
V S Y E V P R L H G D E E R F
1
13132


277
E D F S A S P I F T D T V V F
1
13133


285
F T D T V V F R V A P W I M T
1
13134


301
S T L P P L E V Y V C R V R N
1
13135


318
C F V D A V A E L A R K A G C
1
13136


326
L A R K A G C K L T I C P Q A
1
13137


338
P Q A E N R N D R W I Q D E M
1
13138


340
A E N R N D R W I Q D E M E L
1
13139


367
V F D S P R N G E L Q D F P Y
1
13140


383
R I L G P D F G Y V T R E P R
1
13141


397
R D R S V S G L D S F G N L E
1
13142


408
G N L E V S P P V V A N G K E
1
13143


476
S F V P A P D G K G F R M L L
1
13144


492
S P G A C F K L F Q E K Q K C
1
13145


513
L F Q G V V D D E Q V K T I S
1
13146


575
T E R K K A T A F F P D L V N
1
13147


578
K K A T A F F P D L V N M L V
1
13148


598
G I P K P F G P I I N G C C C
1
13149


616
K V R S L L E P L G L H C T F
1
13150


650
V C R K P F S F K W W N M V P
1
13151







184P1E2 v.2: HLA Peptide


Scoring Results DRB1*0101 15-mers SYFPEITHI










4
V A P W I M T P S T L A P L E
26
13152


3
R V A P W I M T P S T L A P L
23
13153


5
A P W I M T P S T L A P L E V
22
13154


8
I M T P S T L A P L E V Y V C
22
13155


1
V F R V A P W I M T P S T L A
17
13156


6
P W I M T P S T L A P L E V Y
17
13157


11
P S T L A P L E V Y V C R V R
17
13158


13
T L A P L E V Y V C R V R N N
14
13159


2
F R V A P W I M T P S T L A P
12
13160


14
L A P L E V Y V C R V R N N T
9
13161


7
W I M T P S T L A P L E V Y V
8
13162


10
T P S T L A P L E V Y V C R V
8
13163


15
A P L E V Y V C R V R N N T C
7
13164


9
M T P S T L A P L E V Y V C R
2
13165


12
S T L A P L E V Y V C R V R N
1
13166







184P1E2 v.3: HLA Peptide


Scoring Results DRB1*0101 15-mers SYFPEITHI










10
L S F V P V P D G K G F R M L
27
13167


12
F V P V P D G K G F R M L L A
24
13168


3
V G H V D E F L S F V P V P D
22
13169


9
F L S F V P V P D G K G F R M
18
13170


4
G H V D E F L S F V P V P D G
17
13171


6
V D E F L S F V P V P D G K G
16
13172


7
D E F L S F V P V P D G K G F
15
13173


13
V P V P D G K G F R M L L A S
10
13174


14
P V P D G K G F R M L L A S P
10
13175


1
L A V G H V D E F L S F V P V
9
13176


2
A V G H V D E F L S F V P V P
9
13177


5
H V D E F L S F V P V P D G K
9
13178


15
V P D G K G F R M L L A S P G
9
13179


8
E F L S F V P V P D G K G F R
8
13180


11
S F V P V P D G K G F R M L L
2
13181



















TABLE XLIX








SEQ.


Pos
1 2 3 4 5 6 7 8 9 0 1 2 3 4 5
score
ID NO.















184P1E2 v.1: HLA Peptide


Scoring Results DRB1 0301 15-mers SYFPEITHI










268
H V T L L D D S N E D F S A S
37
13182


190
P A A L F D D H K L V L H T S
35
13183


239
V P R L H G D E E R F F V E G
35
13184


514
F Q G V V D D E Q V K T I S I
30
13185


580
A T A F F P D L V N M L V L G
29
13186


116
D I S L D C D L N C E G R Q D
27
13187


226
Y R H V L G Q D K V S Y E V P
27
13188


610
C C C L E E K V R S L L E P L
26
13189


219
P E D V C E A Y R H V L G Q D
25
13190


345
D R W I Q D E M E L G Y V Q A
25
13191


414
P P V V A N G K E Y P L G R I
25
13192


439
G R R V T Q V V R D F L H A Q
25
13193


114
C V D I S L D C D L N C E G R
23
13194


363
T L P V V F D S P R N G E L Q
23
13195


365
P V V F D S P R N G E L Q D F
23
13196


182
V M V L R T Q G P A A L F D D
22
13197


442
V T Q V V R D F L H A Q K V Q
22
13198


486
F R M L L A S P G A C F K L F
22
13199


100
H E P L P L A Y A V L Y L T C
21
13200


227
R H V L G Q D K V S Y E V P R
21
13201


261
F T G L I S F H V T L L D D S
21
13202


281
A S P I F T D T V V F R V A P
21
13203


382
K R I L G P D F G Y V T R E P
21
13204


399
R S V S G L D S F G N L E V S
21
13205


458
P V E L F V D W L A V G H V D
21
13206


562
A E C D I I D I P Q L F K T E
21
13207


76
I I V V M N S P S N D L N D S
20
13208


149
I L L V N C D R D D P S C D V
20
13209


215
H I C G P E D V C E A Y R H V
20
13210


295
P W I M T P S T L P P L E V Y
20
13211


425
L G R I L I G G N L P G S S G
20
13212


466
L A V G H V D E F L S F V P A
20
13213


587
L V N M L V L G K H L G I P K
20
13214


614
E E K V R S L L E P L G L H C
20
13215


15
T S A V C V A G V E T L V D I
19
13216


77
I V V M N S P S N D L N D S H
19
13217


112
L T C V D I S L D C D L N C E
19
13218


170
H V H C L Q D L E D M S V M V
19
13219


174
L Q D L E D M S V M V L R T Q
19
13220


315
N N T C F V D A V A E L A R K
19
13221


323
V A E L A R K A G C K L T I C
19
13222


331
G C K L T I C P Q A E N R N D
19
13223


401
V S G L D S F G N L E V S P P
19
13224


529
N Q V L S N K D L I N Y N K F
19
13225


563
E C D I I D I P Q L F K T E R
19
13226


627
H C T F I D D F T P Y H M L H
19
13227


4
Q R I V R V S L E H P T S A V
18
13228


120
D C D L N C E G R Q D R N F V
18
13229


129
Q D R N F V D K R Q W V W G P
18
13230


180
M S V M V L R T Q G P A A L F
18
13231


317
T C F V D A V A E L A R K A G
18
13232


320
V D A V A E L A R K A G C K L
18
13233


333
K L T I C P Q A E N R N D R W
18
13234


364
L P V V F D S P R N G E L Q D
18
13235


389
F G Y V T R E P R D R S V S G
18
13236


413
S P P V V A N G K E Y P L G R
18
13237


452
A Q K V Q P P V E L F V D W L
18
13238


463
V D W L A V G H V D E F L S F
18
13239


475
L S F V P A P D G K G F R M L
18
13240


476
S F V P A P D G K G F R M L L
18
13241


513
L F Q G V V D D E Q V K T I S
18
13242


545
Q S C I D W N R E V L K R E L
18
13243


569
I P Q L F K T E R K K A T A F
18
13244


623
P L G L H C T F I D D F T P Y
18
13245


626
L H C T F I D D F T P Y H M L
18
13246


30
Y G S V P E G T E M F E V Y G
17
13247


47
G V D I Y I S P N M E R G R E
17
13248


49
D I Y I S P N M E R G R E R A
17
13249


53
S P N M E R G R E R A D T R R
17
13250


58
R G R E R A D T R R W R F D A
17
13251


73
T L E I I V V M N S P S N D L
17
13252


80
M N S P S N D L N D S H V Q I
17
13253


124
N C E G R Q D R N F V D K R Q
17
13254


148
G I L L V N C D R D D P S C D
17
13255


156
R D D P S C D V Q D N C D Q H
17
13256


201
L H T S S Y D A K R A Q V F H
17
13257


253
G L S F P D A G F T G L I S F
17
13258


307
E V Y V C R V R N N T C F V D
17
13259


354
L G Y V Q A P H K T L P V V F
17
13260


446
V R D F L H A Q K V Q P P V E
17
13261


465
W L A V G H V D E F L S F V P
17
13262


496
C F K L F Q E K Q K C G H G R
17
13263


502
E K Q K C G H G R A L L F Q G
17
13264


534
N K D L I N Y N K F V Q S C I
17
13265


548
I D W N R E V L K R E L G L A
17
13266


550
W N R E V L K R E L G L A E C
17
13267


641
H G E V H C G T N V C R K P F
17
13268


238
E V P R L H G D E E R F F V E
16
13269


269
V T L L D D S N E D F S A S P
16
13270


494
G A C F K L F Q E K Q K C G H
16
13271


521
E Q V K T I S I N Q V L S N K
16
13272


543
F V Q S C I D W N R E V L K R
16
13273


566
I I D I P Q L F K T E R K K A
16
13274


570
P Q L F K T E R K K A T A F F
16
13275


67
R W R F D A T L E I I V V M N
15
13276


85
N D L N D S H V Q I S Y H S S
15
13277


130
D R N F V D K R Q W V W G P S
15
13278


308
V Y V C R V R N N T C F V D A
15
13279


479
P A P D G K G F R M L L A S P
15
13280


485
G F R M L L A S P G A C F K L
15
13281


497
F K L F Q E K Q K C G H G R A
15
13282


526
I S I N Q V L S N K D L I N Y
15
13283


528
I N Q V L S N K D L I N Y N K
15
13284


544
V Q S C I D W N R E V L K R E
15
13285


581
T A F F P D L V N M L V L G K
15
13286


600
P K P F G P I I N G C C C L E
15
13287


617
V R S L L E P L G L H C T F I
15
13288


644
V H C G T N V C R K P F S F K
15
13289


59
G R E R A D T R R W R F D A T
14
13290


75
E I I V V M N S P S N D L N D
14
13291


83
P S N D L N D S H V Q I S Y H
14
13292


173
C L Q D L E D M S V M V L R T
14
13293


267
F H V T L L D D S N E D F S A
14
13294


343
R N D R W I Q D E M E L G Y V
14
13295


374
G E L Q D F P Y K R I L G P D
14
13296


386
G P D F G Y V T R E P R D R S
14
13297


509
G R A L L F Q G V V D D E Q V
14
13298


551
N R E V L K R E L G L A E C D
14
13299


552
R E V L K R E L G L A E C D I
14
13300


588
V N M L V L G K H L G I P K P
14
13301


106
A Y A V L Y L T C V D I S L D
13
13302


146
Y G G I L L V N C D R D D P S
13
13303


181
S V M V L R T Q G P A A L F D
13
13304


189
G P A A L F D D H K L V L H T
13
13305


197
H K L V L H T S S Y D A K R A
13
13306


250
F V E G L S F P D A G F T G L
13
13307


266
S F H V T L L D D S N E D F S
13
13308


344
N D R W I Q D E M E L G Y V Q
13
13309


349
Q D E M E L G Y V Q A P H K T
13
13310


372
R N G E L Q D F P Y K R I L G
13
13311


409
N L E V S P P V V A N G K E Y
13
13312


443
T Q V V R D F L H A Q K V Q P
13
13313


520
D E Q V K T I S I N Q V L S N
13
13314


525
T I S I N Q V L S N K D L I N
13
13315


584
F P D L V N M L V L G K H L G
13
13316


636
P Y H M L H G E V H C G T N V
13
13317


3
L Q R I V R V S L E H P T S A
12
13318


17
A V C V A G V E T L V D I Y G
12
13319


22
G V E T L V D I Y G S V P E G
12
13320


24
E T L V D I Y G S V P E G T E
12
13321


39
M F E V Y G T P G V D I Y I S
12
13322


65
T R R W R F D A T L E I I V V
12
13323


96
Y H S S H E P L P L A Y A V L
12
13324


99
S H E P L P L A Y A V L Y L T
12
13325


109
V L Y L T C V D I S L D C D L
12
13326


147
G G I L L V N C D R D D P S C
12
13327


152
V N C D R D D P S C D V Q D N
12
13328


183
M V L R T Q G P A A L F D D H
12
13329


196
D H K L V L H T S S Y D A K R
12
13330


198
K L V L H T S S Y D A K R A Q
12
13331


213
V F H I C G P E D V C E A Y R
12
13332


262
T G L I S F H V T L L D D S N
12
13333


272
L D D S N E D F S A S P I F T
12
13334


294
A P W I M T P S T L P P L E V
12
13335


300
P S T L P P L E V Y V C R V R
12
13336


381
Y K R I L G P D F G Y V T R E
12
13337


407
F G N L E V S P P V V A N G K
12
13338


447
R D F L H A Q K V Q P P V E L
12
13339


456
Q P P V E L F V D W L A V G H
12
13340


468
V G H V D E F L S F V P A P D
12
13341


471
V D E F L S F V P A P D G K G
12
13342


474
F L S F V P A P D G K G F R M
12
13343


487
R M L L A S P G A C F K L F Q
12
13344


530
Q V L S N K D L I N Y N K F V
12
13345


535
K D L I N Y N K F V Q S C I D
12
13346


541
N K F V Q S C I D W N R E V L
12
13347


556
K R E L G L A E C D I I D I P
12
13348


585
P D L V N M L V L G K H L G I
12
13349


589
N M L V L G K H L G I P K P F
12
13350


603
F G P I I N G C C C L E E K V
12
13351


609
G C C C L E E K V R S L L E P
12
13352


618
R S L L E P L G L H C T F I D
12
13353


6
I V R V S L E H P T S A V C V
11
13354


8
R V S L E H P T S A V C V A G
11
13355


19
C V A G V E T L V D I Y G S V
11
13356


23
V E T L V D I Y G S V P E G T
11
13357


29
I Y G S V P E G T E M F E V Y
11
13358


36
G T E M F E V Y G T P G V D I
11
13359


45
T P G V D I Y I S P N M E R G
11
13360


71
D A T L E I I V V M N S P S N
11
13361


74
L E I I V V M N S P S N D L N
11
13362


84
S N D L N D S H V Q I S Y H S
11
13363


91
H V Q I S Y H S S H E P L P L
11
13364


94
I S Y H S S H E P L P L A Y A
11
13365


101
E P L P L A Y A V L Y L T C V
11
13366


102
P L P L A Y A V L Y L T C V D
11
13367


107
Y A V L Y L T C V D I S L D C
11
13368


110
L Y L T C V D I S L D C D L N
11
13369


131
R N F V D K R Q W V W G P S G
11
13370


137
R Q W V W G P S G Y G G I L L
11
13371


159
P S C D V Q D N C D Q H V H C
11
13372


160
S C D V Q D N C D Q H V H C L
11
13373


162
D V Q D N C D Q H V H C L Q D
11
13374


168
D Q H V H C L Q D L E D M S V
11
13375


171
V H C L Q D L E D M S V M V L
11
13376


179
D M S V M V L R T Q G P A A L
11
13377


191
A A L F D D H K L V L H T S S
11
13378


231
G Q D K V S Y E V P R L H G D
11
13379


232
Q D K V S Y E V P R L H G D E
11
13380


247
E R F F V E G L S F P D A G F
11
13381


248
R F F V E G L S F P D A G F T
11
13382


252
E G L S F P D A G F T G L I S
11
13383


260
G F T G L I S F H V T L L D D
11
13384


286
T D T V V F R V A P W I M T P
11
13385


290
V F R V A P W I M T P S T L P
11
13386


298
M T P S T L P P L E V Y V C R
11
13387


299
T P S T L P P L E V Y V C R V
11
13388


305
P L E V Y V C R V R N N T C F
11
13389


310
V C R V R N N T C F V D A V A
11
13390


339
Q A E N R N D R W I Q D E M E
11
13391


368
F D S P R N G E L Q D F P Y K
11
13392


373
N G E L Q D F P Y K R I L G P
11
13393


380
P Y K R I L G P D F G Y V T R
11
13394


392
V T R E P R D R S V S G L D S
11
13395


422
E Y P L G R I L I G G N L P G
11
13396


426
G R I L I G G N L P G S S G R
11
13397


427
R I L I G G N L P G S S G R R
11
13398


431
G G N L P G S S G R R V T Q V
11
13399


450
L H A Q K V Q P P V E L F V D
11
13400


451
H A Q K V Q P P V E L F V D W
11
13401


459
V E L F V D W L A V G H V D E
11
13402


472
D E F L S F V P A P D G K G F
11
13403


483
G K G F R M L L A S P G A C F
11
13404


495
A C F K L F Q E K Q K C G H G
11
13405


523
V K T I S I N Q V L S N K D L
11
13406


532
L S N K D L I N Y N K F V Q S
11
13407


558
E L G L A E C D I I D I P Q L
11
13408


564
C D I I D I P Q L F K T E R K
11
13409


590
M L V L G K H L G I P K P F G
11
13410


594
G K H L G I P K P F G P I I N
11
13411


604
G P I I N G C C C L E E K V R
11
13412


615
E K V R S L L E P L G L H C T
11
13413


621
L E P L G L H C T F I D D F T
11
13414


631
I D D F T P Y H M L H G E V H
11
13415


647
G T N V C R K P F S F K W W N
11
13416


20
V A G V E T L V D I Y G S V P
10
13417


26
L V D I Y G S V P E G T E M F
10
13418


43
Y G T P G V D I Y I S P N M E
10
13419


63
A D T R R W R F D A T L E I I
10
13420


89
D S H V Q I S Y H S S H E P L
10
13421


108
A V L Y L T C V D I S L D C D
10
13422


140
V W G P S G Y G G I L L V N C
10
13423


151
L V N C D R D D P S C D V Q D
10
13424


177
L E D M S V M V L R T Q G P A
10
13425


188
Q G P A A L F D D H K L V L H
10
13426


195
D D H K L V L H T S S Y D A K
10
13427


210
R A Q V F H I C G P E D V C E
10
13428


236
S Y E V P R L H G D E E R F F
10
13429


245
D E E R F F V E G L S F P D A
10
13430


251
V E G L S F P D A G F T G L I
10
13431


280
S A S P I F T D T V V F R V A
10
13432


287
D T V V F R V A P W I M T P S
10
13433


288
T V V F R V A P W I M T P S T
10
13434


303
L P P L E V Y V C R V R N N T
10
13435


316
N T C F V D A V A E L A R K A
10
13436


351
E M E L G Y V Q A P H K T L P
10
13437


353
E L G Y V Q A P H K T L P V V
10
13438


357
V Q A P H K T L P V V F D S P
10
13439


361
H K T L P V V F D S P R N G E
10
13440


378
D F P Y K R I L G P D F G Y V
10
13441


393
T R E P R D R S V S G L D S F
10
13442


398
D R S V S G L D S F G N L E V
10
13443


423
Y P L G R I L I G G N L P G S
10
13444


455
V Q P P V E L F V D W L A V G
10
13445


460
E L F V D W L A V G H V D E F
10
13446


488
M L L A S P G A C F K L F Q E
10
13447


501
Q E K Q K C G H G R A L L F Q
10
13448


503
K Q K C G H G R A L L F Q G V
10
13449


510
R A L L F Q G V V D D E Q V K
10
13450


515
Q G V V D D E Q V K T I S I N
10
13451


522
Q V K T I S I N Q V L S N K D
10
13452


559
L G L A E C D I I D I P Q L F
10
13453


571
Q L F K T E R K K A T A F F P
10
13454


572
L F K T E R K K A T A F F P D
10
13455


573
F K T E R K K A T A F F P D L
10
13456


582
A F F P D L V N M L V L G K H
10
13457


586
D L V N M L V L G K H L G I P
10
13458


596
H L G I P K P F G P I I N G C
10
13459


602
P F G P I I N G C C C L E E K
10
13460


613
L E E K V R S L L E P L G L H
10
13461


619
S L L E P L G L H C T F I D D
10
13462


628
C T F I D D F T P Y H M L H G
10
13463


637
Y H M L H G E V H C G T N V C
10
13464


646
C G T N V C R K P F S F K W W
10
13465


33
V P E G T E M F E V Y G T P G
9
13466


37
T E M F E V Y G T P G V D I Y
9
13467


40
F E V Y G T P G V D I Y I S P
9
13468


54
P N M E R G R E R A D T R R W
9
13469


92
V Q I S Y H S S H E P L P L A
9
13470


122
D L N C E G R Q D R N F V D K
9
13471


135
D K R Q W V W G P S G Y G G I
9
13472


139
W V W G P S G Y G G I L L V N
9
13473


166
N C D Q H V H C L Q D L E D M
9
13474


167
C D Q H V H C L Q D L E D M S
9
13475


178
E D M S V M V L R T Q G P A A
9
13476


203
T S S Y D A K R A Q V F H I C
9
13477


211
A Q V F H I C G P E D V C E A
9
13478


233
D K V S Y E V P R L H G D E E
9
13479


246
E E R F F V E G L S F P D A G
9
13480


256
F P D A C F T C L I S F H V T
9
13481


258
D A G F T G L I S F H V T L L
9
13482


264
L I S F H V T L L D D S N E D
9
13483


276
N E D F S A S P I F T D T V V
9
13484


282
S P I F T D T V V F R V A P W
9
13485


284
I F T D T V V F R V A P W I M
9
13486


306
L E V Y V C R V R N N T C F V
9
13487


350
D E M E L G Y V Q A P H K T L
9
13488


370
S P R N G E L Q D F P Y K R I
9
13489


375
E L Q D F P Y K R I L G P D F
9
13490


376
L Q D F P Y K R I L G P D F G
9
13491


396
P R D R S V S G L D S F G N L
9
13492


404
L D S F G N L E V S P P V V A
9
13493


412
V S P P V V A N G K E Y P L G
9
13494


418
A N G K E Y P L G R I L I G G
9
13495


419
N G K E Y P L G R I L I G G N
9
13496


438
S G R R V T Q V V R D F L H A
9
13497


464
D W L A V G H V D E F L S F V
9
13498


478
V P A P D G K G F R M L L A S
9
13499


489
L L A S P G A C F K L F Q E K
9
13500


527
S I N Q V L S N K D L I N Y N
9
13501


540
Y N K F V Q S C I D W N R E V
9
13502


568
D I P Q L F K T E R K K A T A
9
13503


592
V L G K H L G I P K P F G P I
9
13504


593
L G K H L G I P K P F G P I I
9
13505


608
N G C C C L E E K V R S L L E
9
13506


629
T F I D D F T P Y H M L H G E
9
13507


635
T P Y H M L H G E V H C G T N
9
13508


16
S A V C V A G V E T L V D I Y
8
13509


27
V D I Y G S V P E G T E M F E
8
13510


32
S V P E G T E M F E V Y G T P
8
13511


52
I S P N M E R G R E R A D T R
8
13512


93
Q I S Y H S S H E P L P L A Y
8
13513


145
G Y G G I L L V N C D R D D P
8
13514


163
V Q D N C D Q H V H C L Q D L
8
13515


202
H T S S Y D A K R A Q V F H I
8
13516


212
Q V F H I C G P E D V C E A Y
8
13517


224
E A Y R H V L G Q D K V S Y E
8
13518


230
L G Q D K V S Y E V P R L H G
8
13519


243
H G D E E R F F V E G L S F P
8
13520


244
G D E E R F F V E G L S F P D
8
13521


274
D S N E D F S A S P I F T D T
8
13522


292
R V A P W I M T P S T L P P L
8
13523


304
P P L E V Y V C R V R N N T C
8
13524


321
D A V A E L A R K A G C K L T
8
13525


325
E L A R K A G C K L T I C P Q
8
13526


334
L T I C P Q A E N R N D R W I
8
13527


367
V F D S P R N G E L Q D F P Y
8
13528


403
G L D S F G N L E V S P P V V
8
13529


432
G N L P G S S G R R V T Q V V
8
13530


433
N L P G S S G R R V T Q V V R
8
13531


436
G S S G R R V T Q V V R D F L
8
13532


511
A L L F Q G V V D D E Q V K T
8
13533


531
V L S N K D L I N Y N K F V Q
8
13534


533
S N K D L I N Y N K F V Q S C
8
13535


549
D W N R E V L K R E L G L A E
8
13536


555
L K R E L G L A E C D I I D I
8
13537


561
L A E C D I I D I P Q L F K T
8
13538


576
E R K K A T A F F P D L V N M
8
13539


606
I I N G C C C L E E K V R S L
8
13540


50
I Y I S P N M E R G R E R A D
7
13541


61
E R A D T R R W R F D A T L E
7
13542


118
S L D C D L N C E G R Q D R N
7
13543


121
C D L N C E G R Q D R N F V D
7
13544


123
L N C E G R Q D R N F V D K R
7
13545


204
S S Y D A K R A Q V F H I C G
7
13546


216
I C G P E D V C E A Y R H V L
7
13547


240
P R L H G D E E R F F V E G L
7
13548


335
T I C P Q A E N R N D R W I Q
7
13549


336
I C P Q A E N K N D R W I Q D
7
13550


369
D S P R N G E L Q D F P Y K R
7
13551


391
Y V T R E P R D R S V S G L D
7
13552


411
E V S P P V V A N G K E Y P L
7
13553


445
V V R D F L H A Q K V Q P P V
7
13554


490
L A S P G A C F K L F Q E K Q
7
13555


493
P G A C F K L F Q E K Q K C G
7
13556


507
G H G R A L L F Q G V V D D E
7
13557


516
G V V D D E Q V K T I S I N Q
7
13558


607
I N G C C C L E E K V R S L L
7
13559


128
R Q D R N F V D K R Q W V W G
6
13560


158
D P S C D V Q D N C D Q H V H
6
13561


161
C D V Q D N C D Q H V H C L Q
6
13562


338
P Q A E N R N D R W I Q D E M
6
13563


341
E N R N D R W I Q D E M E L G
6
13564


387
P D F G Y V T R E P R D R S V
6
13565


448
D F L H A Q K V Q P P V E L F
6
13566


537
L I N Y N K F V Q S C I D W N
6
13567


645
H C G T N V C R K P F S F K W
6
13568


650
V C R K P F S F K W W N M V P
6
13569


7
V R V S L E H P T S A V C V A
5
13570


408
G N L E V S P P V V A N G K E
5
13571


424
P L G R I L I G G N L P G S S
5
13572


616
K V R S L L E P L G L H C T F
5
13573


11
L E H P T S A V C V A G V E T
4
13574


70
F D A T L E I I V V M N S P S
4
13575


199
L V L H T S S Y D A K R A Q V
4
13576


293
V A P W I M T P S T L P P L E
4
13577


330
A G C K L T I C P Q A E N R N
4
13578


395
E P R D R S V S G L D S F G N
4
13579


400
S V S G L D S F G N L E V S P
4
13580


406
S F G N L E V S P P V V A N G
4
13581


430
I G G N L P G S S G R R V T Q
4
13582


457
P P V E L F V D W L A V G H V
4
13583


462
F V D W L A V G H V D E F L S
4
13584


508
H G R A L L F Q G V V D D E Q
4
13585


553
E V L K R E L G L A F C D I I
4
13586


583
F F P D L V N M L V L G K H L
4
13587


5
R I V R V S L E H P T S A V C
3
13588


9
V S L E H P T S A V C V A G V
3
13589


46
P G V D I Y I S P N M E R G R
3
13590


69
R F D A T L E I I V V M N S P
3
13591


78
V V M N S P S N D L N D S H V
3
13592


97
H S S H E P L P L A Y A V L Y
3
13593


98
S S H E P L P L A Y A V L Y L
3
13594


105
L A Y A V L Y L T C V D I S L
3
13595


113
T C V D I S L D C D L N C E G
3
13596


115
V D I S L D C D L N C E G R Q
3
13597


119
L D C D L N C E G R Q D R N F
3
13598


141
W G P S G Y G G I L L V N C D
3
13599


143
P S G Y G G I L L V N C D R D
3
13600


175
Q D L E D M S V M V L R T Q G
3
13601


194
F D D H K L V L H T S S Y D A
3
13602


223
C E A Y R H V L G Q D K V S Y
3
13603


235
V S Y E V P R L H G D E E R F
3
13604


237
Y E V P R L H G D E E R F F V
3
13605


296
W I M T P S T L P P L E V Y V
3
13606


302
T L P P L E V Y V C R V R N N
3
13607


319
F V D A V A E L A R K A G C K
3
13608


322
A V A E L A R K A G C K L T I
3
13609


324
A E L A R K A G C K L T I C P
3
13610


348
I Q D E M E L G Y V Q A P H K
3
13611


355
G Y V Q A P H K T L P V V F D
3
13612


358
Q A P H K T L P V V F D S P R
3
13613


360
P H K T L P V V F D S P R N G
3
13614


397
R D R S V S G L D S F G N L E
3
13615


417
V A N G K E Y P L G R I L I G
3
13616


480
A P D G K G F R M L L A S P G
3
13617


506
C G H G R A L L F Q G V V D D
3
13618


546
S C I D W N R E V L K R E L G
3
13619


565
D I I D I P Q L F K T E R K K
3
13620


591
L V L G K H L C I P K P F G P
3
13621


595
K H L G I P K P F G P I I N G
3
13622


597
L G I P K P F G P I I N G C C
3
13623


612
C L E E K V R S L L E P L G L
3
13624


640
L H G E V H C G T N V C R K P
3
13625


649
N V C R K P F S F K W W N M V
3
13626


1
M S L Q R I V R V S L E H P T
2
13627


2
S L Q R I V R V S L E H P T S
2
13628


14
P T S A V C V A G V E T L V D
2
13629


21
A G V E T L V D I Y G S V P E
2
13630


25
T L V D I Y G S V P E G T E M
2
13631


34
P E G T E M F E V Y G T P G V
2
13632


41
E V Y G T P G V D I Y I S P N
2
13633


48
V D I Y I S P N M E R G R E R
2
13634


56
M E R G R E R A D T R R W R F
2
13635


62
R A D T R R W R F D A T L E I
2
13636


72
A T L E I I V V M N S P S N D
2
13637


79
V M N S P S N D L N D S H V Q
2
13638


88
N D S H V Q I S Y H S S H E P
2
13639


90
S H V Q I S Y H S S H E P L P
2
13640


95
S Y H S S H E P L P L A Y A V
2
13641


103
L P L A Y A V L Y L T C V D I
2
13642


111
Y L T C V D I S L D C D L N C
2
13643


136
K R Q W V W G P S G Y G G I L
2
13644


138
Q W V W G P S G Y G G I L L V
2
13645


144
S G Y G G I L L V N C D R D D
2
13646


154
C D R D D P S C D V Q D N C D
2
13647


155
D R D D P S C D V Q D N C D Q
2
13648


176
D L E D M S V M V L R T Q G P
2
13649


192
A L F D D H K L V L H T S S Y
2
13650


220
E D V C E A Y R H V L G Q D K
2
13651


225
A Y R H V L G Q D K V S Y E V
2
13652


242
L H G D E E R F F V E G L S F
2
13653


249
F F V E G L S F P D A G F T G
2
13654


259
A G F T G L I S F H V T L L D
2
13655


265
I S F H V T L L D D S N E D F
2
13656


270
T L L D D S N E D F S A S P I
2
13657


275
S N E D F S A S P I F T D T V
2
13658


285
F T D T V V F R V A P W I M T
2
13659


297
I M T P S T L P P L E V Y V C
2
13660


301
S T L P P L E V Y V C R V R N
2
13661


313
V R N N T C F V D A V A E L A
2
13662


314
R N N T C F V D A V A E L A R
2
13663


318
C F V D A V A E L A R K A G C
2
13664


329
K A G C K L T I C P Q A E N R
2
13665


332
C K L T I C P Q A E N R N D R
2
13666


346
R W I Q D E M E L G Y V Q A P
2
13667


347
W I Q D E M E L G Y V Q A P H
2
13668


359
A P H K T L P V V F D S P R N
2
13669


366
V V F D S P R N G E L Q D F P
2
13670


379
F P Y K R I L G P D F G Y V T
2
13671


383
R I L G P D F G Y V T R E P R
2
13672


388
D F G Y V T R E P R D R S V S
2
13673


390
G Y V T R E P R D R S V S G L
2
13674


410
L E V S P P V V A N G K E Y P
2
13675


415
P V V A N G K E Y P L G R I L
2
13676


420
G K E Y P L G R I L I G G N L
2
13677


421
K E Y P L G R I L I G G N L P
2
13678


429
L I G G N L P G S S G R R V T
2
13679


437
S S G R R V T Q V V R D F L H
2
13680


441
R V T Q V V R D F L H A Q K V
2
13681


444
Q V V R D F L H A Q K V Q P P
2
13682


453
Q K V Q P P V E L F V D W L A
2
13683


454
K V Q P P V E L F V D W L A V
2
13684


469
G H V D E F L S F V P A P D G
2
13685


470
H V D E F L S F V P A P D G K
2
13686


481
P D G K G F R M L L A S P G A
2
13687


482
D G K G F R M L L A S P G A C
2
13688


492
S P G A C F K L F Q E K Q K C
2
13689


519
D D E Q V K T I S I N Q V L S
2
13690


524
K T I S I N Q V L S N K D L I
2
13691


554
V L K R E L G L A E C D I I D
2
13692


557
R E L G L A E C D I I D I P Q
2
13693


567
I D I P Q L F K T E R K K A T
2
13694


577
R K K A T A F F P D L V N M L
2
13695


578
K K A T A F F P D L V N M L V
2
13696


579
K A T A F F P D L V N N L V L
2
13697


598
G I P K P F G P I I N G C C C
2
13698


611
C C L E E K V R S L L E P L G
2
13699


620
L L E P L G L H C T F I D D F
2
13700


622
E P L G L H C T F I D D F T P
2
13701


648
T N V C R K P F S F K W W N M
2
13702


12
E H P T S A V C V A G V E T L
1
13703


18
V C V A G V E T L V D I Y G S
1
13704


28
D I Y G S V P E G T E M F E V
1
13705


31
G S V P E G T E M F E V Y G T
1
13706


38
E M F E V Y G T P G V D I Y I
1
13707


44
G T P G V D I Y I S P N M E R
1
13708


51
Y I S P N M E R G R E R A D T
1
13709


64
D T R R W R F D A T L E I I V
1
13710


68
W R F D A T L E I I V V M N S
1
13711


82
S P S N D L N D S H V Q I S Y
1
13712


125
C E G R Q D R N F V D K R Q W
1
13713


126
E G R Q D R N F V D K R Q W V
1
13714


127
G R Q D R N F V D K R Q W V W
1
13715


132
N F V D K R Q W V W G P S G Y
1
13716


133
F V D K R Q W V W G P S G Y G
1
13717


142
G P S G Y G G I L L V N C D R
1
13718


153
N C D R D D P S C D V Q D N C
1
13719


157
D D P S C D V Q D N C D Q H V
1
13720


164
Q D N C D Q H V H C L Q D L E
1
13721


165
D N C D Q H V H C L Q D L E D
1
13722


169
Q H V H C L Q D L E D M S V M
1
13723


172
H C L Q D L E D M S V M V L R
1
13724


184
V L R T Q G P A A L F D D H K
1
13725


185
L R T Q G P A A L F D D H K L
1
13726


186
R T Q G P A A L F D D H K L V
1
13727


200
V L H T S S Y D A K R A Q V F
1
13728


205
S Y D A K R A Q V F H I C G P
1
13729


206
Y D A K R A Q V F H I C G P E
1
13730


209
K R A Q V F H I C G P E D V C
1
13731


214
F H I C G P E D V C E A Y R H
1
13732


217
C G P E D V C E A Y R H V L G
1
13733


218
G P E D V C E A Y R H V L G Q
1
13734


221
D V C E A Y R H V L G Q D K V
1
13735


229
V L G Q D K V S Y E V P R L H
1
13736


241
R L H G D E E R F F V E G L S
1
13737


254
L S F P D A G F T G L I S F H
1
13738


255
S F P D A G F T G L I S F H V
1
13739


257
P D A G F T G L I S F H V T L
1
13740


263
G L I S F H V T L L D D S N E
1
13741


271
L L D D S N E D F S A S P I F
1
13742


273
D D S N E D F S A S P I F T D
1
13743


277
E D F S A S P I F T D T V V F
1
13744


278
D F S A S P I F T D T V V F R
1
13745


289
V V F R V A P W I M T P S T L
1
13746


309
Y V C R V R N N T C F V D A V
1
13747


311
C R V R N N T C F V D A V A E
1
13748


312
R V R N N T C F V D A V A E L
1
13749


326
L A R K A G C K L T I C P Q A
1
13750


327
A R K A G C K L T I C P Q A E
1
13751


340
A E N R N D R W I Q D E M E L
1
13752


342
N R N D R W I Q D E M E L G Y
1
13753


352
M E L G Y V Q A P H K T L P V
1
13754


362
K T L P V V F D S P R N G E L
1
13755


371
P R N G E L Q D F P Y K R I L
1
13756


394
R E P R D R S V S G L D S F G
1
13757


405
D S F G N L E V S P P V V A N
1
13758


416
V V A N G K E Y P L G R I L I
1
13759


428
I L I G G N L P G S S G R R V
1
13760


435
P G S S G R R V T Q V V R D F
1
13761


440
R R V T Q V V R D F L H A Q K
1
13762


461
L F V D W L A V G H V D E F L
1
13763


467
A V G H V D E F L S F V P A P
1
13764


477
F V P A P D G K G F R M L L A
1
13765


484
K C F R M L L A S P G A C F K
1
13766


491
A S P G A C F K L F Q E K Q K
1
13767


498
K L F Q E K Q K C G H G R A L
1
13768


499
L F Q E K Q K C G H G R A L L
1
13769


500
F Q E K Q K C G H G R A L L F
1
13770


504
Q K C G H G R A L L F Q G V V
1
13771


505
K C G H G R A L L F Q G V V D
1
13772


517
V V D D E Q V K T I S I N Q V
1
13773


536
D L I N Y N K F V Q S C I D W
1
13774


538
I N Y N K F V Q S C I D W N R
1
13775


539
N Y N K F V Q S C I D W N R E
1
13776


542
K F V Q S C I D W N R E V L K
1
13777


574
K T E R K K A T A F F P D L V
1
13778


575
T E R K K A T A F F P D L V N
1
13779


599
I P K P F G P I I N G C C C L
1
13780


625
G L H C T F I D D F T P Y H M
1
13781


630
F I D D F T P Y H M L H G E V
1
13782


633
D F T P Y H M L H G E V H C G
1
13783


634
F T P Y H M L H G E V H C G T
1
13784


638
H M L H G E V H C G T N V C R
1
13785


639
M L H G E V H C G T N V C R K
1
13786


643
E V H C G T N V C R K P F S F
1
13787







184P1E2 v.2: HLA Peptide


Scoring Results DRB1 0301 15-mers SYFPEITHI










6
P W I M T P S T L A P L E V Y
21
13788


5
A P W I M T P S T L A P L E V
13
13789


11
P S T L A P L E V Y V C R V R
12
13790


1
V F R V A P W I M T P S T L A
11
13791


9
M T P S T L A P L E V Y V C R
11
13792


10
T P S T L A P L E V Y V C R V
10
13793


14
L A P L E V Y V C R V R N N T
10
13794


3
R V A P W I M T P S T L A P L
8
13795


15
A P L E V Y V C R V R N N T C
8
13796


4
V A P W I M T P S T L A P L E
4
13797


7
W I M T P S T L A P L E V Y V
3
13798


13
T L A P L E V Y V C R V R N N
3
13799


12
S T L A P L E V Y V C R V R N
2
13800


8
I M T P S T L A P L E V Y V C
1
13801







184P1E2 v.3: HLA Peptide


Scoring Results DRB1 0301 15-mers SYFPEITHI










sq
L A V G H V D E F L S F V P V
20
13802


11
S F V P V P D G K G F R M L L
19
13803


10
L S F V P V P D G K G F R M L
18
13804


14
P V P D G K G F R M L L A S P
15
13805


3
V G H V D E F L S F V P V P D
12
13806


6
V D E F L S F V P V P D G K G
12
13807


9
F L S F V P V P D G K G F R M
12
13808


7
D E F L S F V P V P D G K G F
11
13809


12
F V P V P D G K G F R M L L A
11
13810


13
V P V P D G K G F R M L L A S
9
13811


15
V P D G K G F R M L L A S P G
3
13812


4
G H V D E F L S F V P V P D G
2
13813


2
A V G H V D E F L S F V P V P
1
13814


5
H V D E F L S F V P V P D G K
1
13815



















TABLE L








SEQ.





ID


Pos
1 2 3 4 5 6 7 8 9 0 1 2 3 4 5
score
NO.















184P1E2 v.1: HLA Peptide Scoring Results DRBI 0401


15-mers SYFPEITHI










65
T R R W R F D A T L E I I V V
28
13816


258
D A G F T G L I S F H V T L L
28
13817


353
E L G Y V Q A P H K T L P V V
28
13818


537
L I N Y N K F V Q S C I D W N
28
13819


581
T A F F P D L V N M L V L G K
28
13820


73
T L E I I V V M N S P S N D L
26
13821


219
P E D V C E A Y R H V L G Q D
26
13822


389
F G Y V T R E P R D R S V S G
26
13823


398
D R S V S G L D S F G N L E V
26
13824


401
V S G L D S F G N L E V S P P
26
13825


439
G R R V T Q V V R D F L H A Q
26
13826


443
T Q V V R D F L H A Q K V Q P
26
13827


514
F Q G V V D D E Q V K T I S I
26
13828


563
E C D I I D I P Q L F K T E R
26
13829


637
Y H M L H G E V H C G T N V C
26
13830


37
T E M F E V Y G T P G V D I Y
22
13831


104
P L A Y A V L Y L T C V D I S
22
13832


223
C E A Y R H V L G Q D K V S Y
22
13833


253
G L S F P D A C F T C L I S F
22
13834


288
T V V F R V A P W I M T P S T
22
13835


316
N T C F V D A V A E L A R K A
22
13836


365
P V V F D S P R N G E L Q D F
22
13837


386
G P D F G Y V T R E P R D R S
22
13838


462
F V D W L A V G H V D E F L S
22
13839


494
G A C F K L F Q E K Q K C G H
22
13840


511
A L L F Q G V V D D E Q V K T
22
13841


570
P Q L F K T E R K K A T A F F
22
13842


580
A T A F F P D L V N M L V L G
22
13843


600
P K P F G P I I N G C C C L E
22
13844


627
H C T F I D D F T P Y H M L H
22
13845


6
I V R V S L E H P T S A V C V
20
13846


15
T S A V C V A C V E T L V D I
20
13847


17
A V C V A G V E T L V D I Y C
20
13848


45
T P C V D I Y I S P N M E R G
20
13849


71
D A T L E I I V V M N S P S N
20
13850


74
L F I I V V M N S P S N D L N
20
13851


76
I I V V M N S P S N D L N D S
20
13852


77
I V V M N S P S N D L N D S H
20
13853


102
P L P L A Y A V L Y L T C V D
20
13854


106
A Y A V L Y L T C V D I S L D
20
13855


116
D I S L D C D L N C E G R Q D
20
13856


120
D C D L N C E G R Q D R N F V
20
13857


149
I L L V N C D R D D P S C D V
20
13858


171
V H C L Q D L E D N S V N V B
20
13859


179
D M S V M V L R T Q G P A A L
20
13860


190
P A A L F D D H K L V L H T S
20
13861


197
H K L V L H T S S Y D A K R A
20
13862


248
R F F V E G L S F P D A G F T
20
13863


261
F T G L I S F H V T L L D D S
20
13864


268
H V T L L D D S N E D F S A S
20
13865


287
D T V V F R V A P W I M T P S
20
13866


307
E V Y V C R V R N N T C F V D
20
13867


320
V D A V A E L A R K A G C K L
20
13868


331
G C K L T I C P Q A E N R N D
20
13869


351
E M E L G Y V Q A P H K T L P
20
13870


373
N G E L Q D F P Y K R I L G P
20
13871


382
K R I L G P D F G Y V T R E P
20
13872


422
E Y P L G R I L I C G N L P C
20
13873


442
V T Q V V R D F L H A Q K V Q
20
13874


456
Q P P V E L F V D W L A V G H
20
13875


460
E L F V D W L A V G H V D E F
20
13876


465
W L A V C H V D E F L S F V P
20
13877


468
V C H V D E F L S F V P A P D
20
13878


496
C F K L F Q E K Q K C G H G R
20
13879


515
Q G V V D D E Q V K T I S I N
20
13880


520
D E Q V K T I S I N Q V L S N
20
13881


525
T I S I N Q V L S N K D L I N
20
13882


556
K R S L G L A E C D I I D I P
20
13883


569
I P Q L F K T E R K K A T A F
20
13884


588
V N M L V L G K H L G I P K P
20
13885


618
R S L L S P L C L H C T F I D
20
13886


628
C T F I D D F T P Y H M L H G
20
13887


5
R I V R V S L E H P T S A V C
18
13888


83
P S N D L N D S H V Q I S Y H
18
13889


129
Q D R N F V D K R Q W V W C P
18
13890


152
V N C D R D D P S C D V Q D N
18
13891


173
C L Q D L E D M S V M V L R T
18
13892


189
G P A A L F D D H K L V L H T
18
13893


194
F D D H K L V L H T S S Y D A
18
13894


202
H P 5 S Y D A K R A Q V F H I
18
13895


224
E A Y R H V L C Q D K V S Y E
18
13896


240
P R L H G D E E R F F V E C L
18
13897


278
D F S A S P I F T D T V V F R
18
13898


291
F R V A P W I M T P S T L P P
18
13899


304
P P L E V Y V C R V R N N T C
18
13900


362
K T L P V V F D S P R N C E L
18
13901


405
D S F G N L E V S P P V V A N
18
13902


419
N G K E Y P L G R I L I G G N
18
13903


484
K G F R M L L A S P G A C F K
18
13904


493
P C A C F K L F Q E K Q K C G
18
13905


507
G H G R A L L F Q G V V D D E
18
13906


517
V V D D S Q V K T I S I N Q V
18
13907


522
Q V K T I S I N Q V L S N K D
18
13908


526
I S I N Q V L S N K D L I N Y
18
13909


531
V L S N K D L I N Y N K F V Q
18
13910


543
F V Q S C I D W N R E V L K R
18
13911


567
I D I P Q L F K T E R K K A T
18
13912


306
L E V Y V C R V R N N T C F V
17
13913


27
V D I Y G S V P E G T E M F E
16
13914


67
R W R F D A T L E I I V V M N
16
13915


108
A V L Y L T C V D I S L D C D
16
13916


136
K R Q W V W G P S G Y G G I L
16
13917


138
Q W V W G P S G Y G G I L L V
16
13918


143
P S G Y G G I L L V N C D R D
16
13919


211
A Q V F H I C G P E D V C E A
16
13920


246
E E R F F V E G L S F P D A G
16
13921


247
E R F F V E G L S F P D A G F
16
13922


264
L I S F H V T L L D D S N E D
16
13923


282
S P I F T D T V V F R V A P W
16
13924


293
V A P W I M T P S T L P P L E
16
13925


344
N D R W I Q D E M E L G Y V Q
16
13926


378
D F P Y K R I L G P D F G Y V
16
13927


404
L D S F G N L E V S P P V V A
16
13928


446
V R D F L H A Q K V Q P P V E
16
13929


471
V D E F L S F V P A P D G K C
16
13930


474
F L S F V P A P D G K G F R M
16
13931


483
G K G F R M L L A S P G A C F
16
13932


540
Y N K F V Q S C I D W N R E V
16
13933


547
C I D W N R E V L K R E L G L
16
13934


634
F T P Y H M L H G E V H C G T
16
13935


180
M S V M V L R T Q G P A A L F
15
13936


610
C C C L E E K V R S L L E P L
15
13937


3
L Q R I V R V S L E H P T S A
14
13938


4
Q R I V R V S L E H P T S A V
14
13939


8
R V S L E H P T S A V C V A G
14
13940


20
V A G V E T L V D I Y G S V P
14
13941


23
V E T L V D I Y G S V P E G T
14
13942


24
E T L V D I Y G S V P E G T E
14
13943


26
L V D I Y G S V P E G T E M F
14
13944


36
G T E M F E V Y G T P G V D I
14
13945


39
M F E V Y G T P G V D I Y I S
14
13946


47
G V D I Y I S P N M E R G R E
14
13947


84
S N D L N D S H V Q I S Y H S
14
13948


89
D S H V Q I S Y H S S H E P L
14
13949


91
H V Q I S Y H S S H E P L P L
14
13950


100
H E P L P L A Y A V L Y L T C
14
13951


107
Y A V L Y L T C V D I S L D C
14
13952


109
V L Y L T C V D I S B D C D L
14
13953


112
L T C V D I S L D C D L N C E
14
13954


114
C V D I S L D C D L N C E G R
14
13955


146
Y G G I L L V N C D R D D P S
14
13956


148
G I L L V N C D R D D P S C D
14
13957


160
S C D V Q D N C D Q H V H C L
14
13958


168
D Q H V H C L Q D L E D M S V
14
13959


174
L Q D L E D M S V M V L R T Q
14
13960


177
L E D M S V M V L R T Q G P A
14
13961


181
S V M V L R T Q G P A A L F D
14
13962


182
V M V L R T Q G P A A L F D D
14
13963


196
D H K L V L H T S S Y D A K R
14
13964


198
K L V L H T S S Y D A K R A Q
14
13965


210
R A Q V F H I C G P E D V C E
14
13966


227
R H V L G Q D K V S Y E V P R
14
13967


232
Q D K V S Y E V P R L H G D E
14
13968


236
S Y E V P R L H G D E E R F F
14
13969


239
V P R L H G D E E R F F V E G
14
13970


251
V E G L S F P D A G F T G L I
14
13971


262
T G L I S F H V T L L D D S N
14
13972


266
S F H V T L L D D S N E D F S
14
13973


269
V T L L D D S N E D F S A S P
14
13974


281
A S P I F T D T V V F R V A P
14
13975


290
V F R V A P W I H T P S T L P
14
13976


294
A P W I M T P S T L P P L E V
14
13977


295
P W I N T P S T L P P L E V Y
14
13978


300
P S T L P P L E V Y V C R V R
14
13979


305
P L E V Y V C R V R N N T C F
14
13980


317
T C F V D A V A E L A R K A G
14
13981


333
K L T I C P Q A E N R N D R W
14
13982


345
D R W I Q D E M E L G Y V Q A
14
13983


361
H K T L P V V F D S P K N G E
14
13984


363
T L P V V F D S P R N G E L Q
14
13985


364
L P V V F D S P R N G E L Q D
14
13986


381
Y K R I L G P D F G Y V T R E
14
13987


407
F G N L E V S P P V V A N C K
14
13988


414
P P V V A N G K E Y P L C K I
14
13989


425
L G R I L I G G N L P G S S G
14
13990


426
G K I L I G G N L P G S S G R
14
13991


427
R I L I C G N L P C S S G R R
14
13992


431
G G N L P G S S G R R V T Q V
14
13993


452
A Q K V Q P P V E L F V D W L
14
13994


458
P V E L F V D W L A V C H V D
14
13995


472
D E F L S F V P A P D G K G F
14
13996


485
G F R M L L A S P C A C F K L
14
13997


487
R M L L A S P C A C F K L F Q
14
13998


523
V K T I S I N Q V L S N K D B
14
13999


528
I N Q V L S N K D L I N Y N K
14
14000


534
N K D L I N Y N K F V Q S C I
14
14001


535
K D L I N Y N K F V Q S C I D
14
14002


545
Q S C I D W N R E V L K R E L
14
14003


552
R E V L K R E L G L A E C D I
14
14004


558
E L G L A E C D I I D I P Q L
14
14005


564
C D I I D I P Q L F K T E R K
14
14006


566
I T D I P Q L F K T E R K K A
14
14007


584
F P D L V N M L V L G K H L G
14
14008


585
P D L V N M L V L G K H L G I
14
14009


587
L V N M L V L G K H L G I P K
14
14010


590
M L V L G K H L G I P K P F G
14
14011


596
H L G I P K P F G P I I N C C
14
14012


614
E E K V R S L L E P L G L H C
14
14013


621
L E P L G L H C T F I D D F T
14
14014


641
H G E V H C G T N V C R K P F
14
14015


7
V R V S L E H P T S A V C V A
12
14016


12
E H P T S A V C V A G V E T L
12
14017


14
P T S A V C V A G V E T L V D
12
14018


18
V C V A G V E T L V D I Y G S
12
14019


21
A G V E T L V D I Y G S V P E
12
14020


22
G V E T L V D I Y G S V P E G
12
14021


29
I Y G S V P E G T E M F E V Y
12
14022


34
P E G T E M F E V Y G T P G V
12
14023


42
V Y G T P G V D I Y I S P N M
12
14024


44
G T P G V D I Y I S P N M E R
12
14025


57
E R G R E R A D T R R W R F D
12
14026


58
R G R E R A D T R R W R F D A
12
14027


66
R R W R F D A T L E I I V V M
12
14028


68
W R F D A T L E I I V V M N S
12
14029


69
R F D A T L E I I V V M N S P
12
14030


70
F D A T L E I I V V N N S P S
12
14031


80
N N S P S N D L N D S H V Q I
12
14032


81
N S P S N D L N D S H V Q I S
12
14033


82
S P S N D L N D S H V Q I S Y
12
14034


87
L N D S H V Q I S Y H S S H E
12
14035


88
N D S H V Q I S Y H S S H E P
12
14036


92
V Q I S Y H S S H E P L P L A
12
14037


95
S Y H S S H E P L P L A Y A V
12
14038


97
H S S H E P L P L A Y A V L Y
12
14039


99
S H E P L P L A Y A V L Y L T
12
14040


110
L Y L T C V D I S L D C D L N
12
14041


113
T C V D I S L D C D L N C F C
12
14042


121
C D L N C E G R Q D R N F V D
12
14043


123
L N C E G R Q D R N F V D K R
12
14044


124
N C E G R Q D R N F V D K R Q
12
14045


127
G R Q D R N F V D K R Q W V W
12
14046


145
G Y G G I L L V N C D R D D P
12
14047


156
R D D P S C D V Q D N C D Q H
12
14048


158
D P S C D V Q D N C D Q H V H
12
14049


162
D V Q D N C D Q H V H C L Q D
12
14050


164
Q D N C D Q H V H C L Q D L E
12
14051


170
H V H C L Q D L E D M S V M V
12
14052


172
H C L Q D L E D M S V M V L R
12
14053


176
D L E D M S V M V L R T Q G P
12
14054


178
E D M S V M V L R T Q G P A A
12
14055


187
T Q G P A A L F D D H K L V L
12
14056


188
Q G P A A L F D D H K L V L H
12
14057


193
L F D D H K L V L H T S S Y D
12
14058


195
D D H K L V L H T S S Y D A K
12
14059


201
L H T S S Y D A K R A Q V F H
12
14060


205
S Y D A K R A Q V F H I C G P
12
14061


207
D A K R A Q V F H I C G P E D
12
14062


214
F H I C G P E D V C E A Y R H
12
14063


218
G P E D V C E A Y R H V L G Q
12
14064


233
D K V S Y E V P R L H G D E E
12
14065


243
H G D E E R F F V E G L S F P
12
14066


244
G D E E R F F V E G L S F P D
12
14067


254
L S F P D A G F T G L I S F H
12
14068


255
S F P D A G F T G L I S F H V
12
14069


259
A G F T G L I S F H V T L L D
12
14070


260
G F T G L I S F H V T L L D D
12
14071


265
I S F H V T L L D D S N E D F
12
14072


267
F H V T L L D D S N E D F S A
12
14073


271
L L D D S N E D F S A S P I F
12
14074


272
L D D S N E D F S A S P I F T
12
14075


273
D D S N E D F S A S P T F T D
12
14076


275
S N E D F S A S P I F T D T V
12
14077


279
F S A S P I F T D T V V F R V
12
14078


283
P I F T D T V V F R V A P W I
12
14079


284
I F T D T V V F R V A P W I M
12
14080


297
I M T P S T L P P L E V Y V C
12
14081


302
T L P P L E V Y V C R V R N N
12
14082


309
Y V C R V R N N T C F V D A V
12
14083


313
V R N N T C F V D A V A E L A
12
14084


315
N N T C F V D A V A E L A R K
12
14085


318
C F V D A V A E L A R K A G C
12
14086


319
F V D A V A E L A R K A G C K
12
14087


324
A F L A R K A G C K L T I C P
12
14088


328
R K A G C K L T I C P Q A F N
12
14089


334
L T I C P Q A E N R N D R W I
12
14090


335
T I C P Q A E N R N D R W I Q
12
14091


338
P Q A E N R N D R W I Q D E M
12
14092


341
E N R N D R W I Q D E M E L G
12
14093


347
W I Q D E M E L G Y V Q A P H
12
14094


348
I Q D E N E L G Y V Q A P H K
12
14095


355
G Y V Q A P H K T L P V V F D
12
14096


369
D S P R N G E L Q D F P Y K R
12
14097


392
V T R E P R D R S V S G L D S
12
14098


395
E P R D R S V S G L D S F G N
12
14099


399
R S V S G L D S F G N L E V S
12
14100


406
S F G N L E V S P P V V A N G
12
14101


410
L S V S P P V V A N G K S Y P
12
14102


411
E V S P P V V A N G K E Y P L
12
14103


423
Y P L G R I L I G C N L P G S
12
14104


432
G N L P G S S G R R V T Q V V
12
14105


433
N L P G S S G R R V T Q V V R
12
14106


436
G S S G R R V T Q V V R D F L
12
14107


444
Q V V R D F L H A Q K V Q P P
12
14108


445
V V R D F L H A Q K V Q P P V
12
14109


455
V Q P P V E L F V D W L A V G
12
14110


457
P P V E L F V D W L A V G H V
12
14111


467
A V G H V D E F L S F V P A P
12
14112


469
G H V D E F L S F V P A P D G
12
14113


479
P A P D G K G F R M L L A S P
12
14114


480
A P D G K G F R M L L A S P G
12
14115


500
F Q E K Q K C G H G R A L L F
12
14116


502
E K Q K C G H G R A L L F Q G
12
14117


506
C G H G R A L L F Q G V V D D
12
14118


512
L L F Q G V V D D E Q V K T I
12
14119


513
L F Q G V V D D E Q V K T I S
12
14120


519
D D E Q V K T I S I N Q V L S
12
14121


521
E Q V K T I S I N Q V L S N K
12
14122


530
Q V L S N K D L I N Y N K F V
12
14123


533
S N K D L I N Y N K F V Q S C
12
14124


538
I N Y N K F V Q S C I D W N R
12
14125


544
V Q S C I D W N R E V L K R E
12
14126


548
I D W N R E V L K R E L G L A
12
14127


549
D W N R E V L K R E L G L A S
12
14128


555
L K R E L G L A E C D I I D I
12
14129


560
G L A E C D I I D I P Q L F K
12
14130


561
L A E C D I I D I P Q L F K T
12
14131


576
E R K K A T A F F P D L V N M
12
14132


577
R K K A T A F F P D L V N M L
12
14133


578
K K A T A F F P D L V N M L V
12
14134


582
A F F P D L V N M L V L G K H
12
14135


586
D L V N M L V L G K H L C I P
12
14136


597
L G I P K P F G P I I N G C C
12
14137


608
N G C C C L E E K V R S L L E
12
14138


609
G C C C L E E K V R S L L E P
12
14139


611
C C L E E K V R S L L E P L G
12
14140


615
E K V R S L L E P L G L H C T
12
14141


620
L L E P L G L H C T F I D D F
12
14142


624
L G L H C T F I D D F T P Y H
12
14143


633
D F T P Y H M L H G E V H C G
12
14144


638
H M L H G E V H C G T N V C R
12
14145


644
V H C G T N V C R K P F S F K
12
14146


130
D R N F V D K R Q W V W G P S
11
14147


40
F E V Y G T P G V D I Y I S P
10
14148


48
V D I Y I S P N M E R G R E R
10
14149


93
Q I S Y H S S H E P L P L A Y
10
14150


191
A A L F D D H K L V L H T S S
10
14151


234
K V S Y S V P R L H G D E E R
10
14152


276
N S D F S A S P I F T D T V V
10
14153


420
G K E Y P L G R I L I G G N L
10
14154


459
V E L F V D W L A V G H V D E
10
14155


631
I D D F T P Y H M L H G E V H
10
14156


286
T D T V V F R V A P W I M T P
9
14157


323
V A E L A R K A G C K L T I C
9
14158


551
N R E V L K R E L G L A E C D
9
14159


647
G T N V C R K P F S F K W W N
9
14160


30
Y G S V P E G T E M F E V Y G
8
14161


49
D I Y I S P N M E R G R E R A
8
14162


53
S P N M E R G R E R A D T R R
8
14163


75
E I I V V M N S P S N D L N D
8
14164


137
R Q W V W G P S G Y G G I L L
8
14165


147
G G I L L V N C D R D D P S C
8
14166


213
V F H I C G P E D V C E A Y R
8
14167


226
Y R H V L G Q D K V S Y E V P
8
14168


303
L P P L E V Y V C R V R N N T
8
14169


310
V C R V R N N T C F V D A V A
8
14170


349
Q D E M E L G Y V Q A P H K T
8
14171


354
L G Y V Q A P H K T L P V V F
8
14172


409
N L E V S P P V V A N G K E Y
8
14173


413
S P P V V A N G K E Y P L G R
8
14174


447
R D F L H A Q K V Q P P V E L
8
14175


463
V D W L A V G H V D E F L S F
8
14176


475
L S F V P A P D G K G F R M L
8
14177


486
F R M L L A S P G A C F K L F
8
14178


509
G R A L L F Q G V V D D E Q V
8
14179


510
R A L L F Q G V V D D E Q V K
8
14180


541
N K F V Q S C I D W N R E V L
8
14181


594
G K H L G I P K P F G P I I N
8
14182


603
F C P I I N G C C C L S E K V
8
14183


604
G P I I N G C C C L E E K V R
8
14184


617
V R S L L E P L G L H C T F I
8
14185


623
P L G L H C T F I D D F T P Y
8
14186


636
P Y H M L H G S V H C G T N V
8
14187


52
I S P N M E R G R S R A D T R
7
14188


54
P N M E R G R E R A D T R R W
7
14189


61
E R A D T R R W R F D A T L E
7
14190


228
H V L G Q D K V S Y S V P R L
7
14191


391
Y V T R E P R D R S V S G L D
7
14192


435
P G S S G R R V T Q V V R D F
7
14193


448
D F L H A Q K V Q p p v S L F
7
14194


1
M S L Q R I V R V S L E H P T
6
14195


9
V S L E H P T S A V C V A G V
6
14196


10
S L E H P T S A V C V A G V E
6
14197


11
L E H P T S A V C V A G V E T
6
14198


16
S A V C V A G V E T L V D I Y
6
14199


28
D I Y G S V P 5 G T E M F E V
6
14200


31
G S V P E G T E M F E V Y G T
6
14201


32
S V P E G T E M F E V Y G T P
6
14202


35
E G T E M F E V Y G T P G V D
6
14203


41
E V Y G T P G V D I Y I S P N
6
14204


43
Y G T P G V D I Y I S P N M E
6
14205


46
P G V D I Y I S P N M E R G R
6
14206


50
I Y I S P N M E R G R E R A D
6
14207


51
Y I S P N M E R G R E R A D T
6
14208


55
N M E R G R E R A D T R R W R
6
14209


59
G R E R A D T R R W R F D A T
6
14210


62
R A D T R R W R F D A T L E I
6
14211


64
D T R R W R F D A T L B I I V
6
14212


72
A T L E I I V V M N S P S N D
6
14213


79
V M N S P S N D L N D S H V Q
6
14214


85
N D L N D S H V Q I S Y H S S
6
14215


86
D L N D S H V Q I S Y H S S H
6
14216


90
S H V Q I S Y H S S H E P L P
6
14217


94
I S Y H S S H E P L P L A Y A
6
14218


98
S S H E P L P L A Y A V L Y L
6
14219


101
E P L P L A Y A V L Y L T C V
6
14220


103
L P L A Y A V L Y L T C V D I
6
14221


105
L A Y A V L Y L T C V D I S L
6
14222


111
Y L T C V D I S L D C D L N C
6
14223


117
I S L D C D L N C E G R Q D R
6
14224


118
S L D C D L N C E G R Q D R N
6
14225


128
R Q D R N F V D K R Q W V W G
6
14226


133
F V D K R Q W V W G P S G Y G
6
14227


134
V D K R Q W V W G P S G Y G G
6
14228


139
W V W G P S G Y G G I L L V N
6
14229


140
V W G P S G Y G G I L L V N C
6
14230


141
W G P S G Y G G I L L V N C D
6
14231


142
G P S G Y G G I L L V N C D R
6
14232


144
S G Y G G I L L V N C D R D D
6
14233


151
L V N C D R D D P S C D V Q D
6
14234


153
N C D R D D P S C D V Q D N C
6
14235


155
D R D D P S C D V Q D N C D Q
6
14236


157
D D P S C D V Q D N C D Q H V
6
14237


159
P S C D V Q D N C D Q H V H C
6
14238


161
C D V Q D N C D Q H V H C L Q
6
14239


163
V Q D N C D Q H V H C L Q D L
6
14240


165
D N C D Q H V H C L Q D L E D
6
14241


166
N C D Q H V H C L Q D L E D M
6
14242


167
C D Q H V H C L Q D L E D M S
6
14243


169
Q H V H C L Q D L E D M S V M
6
14244


185
L R T Q G P A A L F D D H K L
6
14245


186
R T Q G P A A L F D D H K L V
6
14246


200
V L H T S S Y D A K R A Q V F
6
14247


208
A K R A Q V F H I C G P E D V
6
14248


215
H I C G P E D V C E A Y R H V
6
14249


216
I C G P E D V C E A Y R H V L
6
14250


220
E D V C E A Y R H V L G Q D K
6
14251


225
A Y R H V L G Q D K V S Y E V
6
14252


229
V L G Q D K V S Y E V P R L H
6
14253


231
G Q D K V S Y E V P R L H G D
6
14254


241
R L H G D E E R F F V E G L S
6
14255


245
D E E R F F V E G L S F P D A
6
14256


250
F V E G L S F P D A G F T G L
6
14257


252
E G L S F P D A G F T G L I S
6
14258


263
G L I S F H V T L L D D S N E
6
14259


270
T L L D D S N E D F S A S P I
6
14260


274
D S N E D F S A S P I F T D T
6
14261


277
E D F S A S P I F T D T V V F
6
14262


280
S A S P I F T D T V V F R V A
6
14263


285
F T D T V V F R V A P W I M T
6
14264


292
R V A P W I M T P S T L P P L
6
14265


299
T P S T L P P L E V Y V C R V
6
14266


301
S T L P P L E V Y V C R V R N
6
14267


311
C R V R N N T C F V D A V A E
6
14268


312
R V R N N T C F V D A V A E L
6
14269


314
R N N T C F V D A V A E L A R
6
14270


321
D A V A E L A R K A G C K L T
6
14271


325
E L A R K A G C K L T I C P Q
6
14272


329
K A G C K L T I C P Q A E N R
6
14273


330
A G C K L T I C P Q A E N R N
6
14274


336
I C P Q A E N R N D R W I Q D
6
14275


339
Q A E N R N D R W I Q D E M E
6
14276


342
N R N D R W I Q D E M E L G Y
6
14277


343
R N D R W I Q D E M E L G Y V
6
14278


346
R W I Q D E M E L G Y V Q A P
6
14279


350
D E M E L G Y V Q A P H K T L
6
14280


352
M B L G Y V Q A P H K T L P V
6
14281


358
Q A P H K T L P V V F D S P R
6
14282


359
A P H K T L P V V F D S P R N
6
14283


360
P H K T L P V V F D S P R N C
6
14284


370
S P R N C E L Q D F P Y K R I
6
14285


371
P R N G E L Q D F P Y K R I L
6
14286


372
R N G E L Q D F P Y K R I L G
6
14287


374
G F L Q D F P Y K R I L G P D
6
14288


375
E L Q D F P Y K R I L G P D F
6
14289


379
F P Y K R I L G P D F G Y V T
6
14290


383
R I L G P D F G Y V T R E P R
6
14291


385
L G P D F G Y V T R E P R D R
6
14292


390
G Y V T R E P R D R S V S G L
6
14293


394
R E P R D R S V S G L D S F G
6
14294


402
S G L D S F G N L E V S P P V
6
14295


408
G N L E V S P P V V A N G K E
6
14296


412
V S P P V V A N G K E Y P L G
6
14297


416
V V A N G K E Y P L G R I L I
6
14298


417
V A N G K E Y P L G R I L I G
6
14299


418
A N G K E Y P L G R I L I G G
6
14300


424
P L G R I L I G G N L P G S S
6
14301


428
I L I G G N L P G S S G R R V
6
14302


429
L I G G N L P G S S G R R V T
6
14303


430
I G G N L P G S S G R R V T Q
6
14304


440
R R V T Q V V R D F L H A Q K
6
14305


449
F L H A Q K V Q P P V E L F V
6
14306


450
L H A Q K V Q P P V E L F V D
6
14307


453
Q K V Q P P V E L F V D W L A
6
14308


454
K V Q P P V E L F V D W L A V
6
14309


461
L F V D W L A V G H V D E F L
6
14310


464
D W L A V G H V D E F L S F V
6
14311


466
L A V G H V D E F L S F V P A
6
14312


473
E F L S F V P A P D G K G F R
6
14313


476
S F V P A P D G K G F R M L L
6
14314


477
F V P A P D G K G F R M L L A
6
14315


488
M L L A S P G A C F K L F Q E
6
14316


489
L L A S P G A C F K L F Q E K
6
14317


490
L A S P G A C F K L F Q E K Q
6
14318


491
A S P G A C F K L F Q E K Q K
6
14319


501
Q E K Q K C G H G R A L L F Q
6
14320


505
K C G H G R A L L F Q G V V D
6
14321


508
H G R A L L F Q G V V D D E Q
6
14322


516
G V V D D E Q V K T I S I N Q
6
14323


524
K T I S I N Q V L S N K D L I
6
14324


527
S I N Q V L S N K D L I N Y N
6
14325


532
L S N K D L I N Y N K F V Q S
6
14326


542
K F V Q S C I D W N R E V L K
6
14327


553
E V L K R E L G L A E C D I I
6
14328


557
R E L G L A E C D I I D I P Q
6
14329


559
L C L A E C D I I D I P Q L F
6
14330


562
A E C D I I D I P Q L F K T E
6
14331


574
K T E R K K A T A F F P D L V
6
14332


579
K A T A F F P D L V N M L V L
6
14333


591
L V L G K H L G I P K P F G P
6
14334


593
L G K H L G I P K P F G P I I
6
14335


601
K P F G P I I N G C C C L E E
6
14336


602
P F G P I I N G C C C L E E K
6
14337


605
P I I N G C C C L E E K V R S
6
14338


606
I I N G C C C L E E K V R S L
6
14339


607
I N G C C C L E E K V R S L L
6
14340


613
L E E K V R S L L E P L G L H
6
14341


616
K V R S L L E P L G L H C T F
6
14342


619
S L L E P L G L H C T F I D D
6
14343


625
G L H C T F I D D F T P Y H M
6
14344


626
L H C T F I D D F T P Y H M L
6
14345


629
T F I D D F T P Y H M L H G E
6
14346


630
F I D D F T P Y H M L H G E V
6
14347


639
M L H G E V H C G T N V C R K
6
14348


640
L H G E V H C G T N V C R K P
6
14349


642
G E V H C G T N V C R K P F S
6
14350


645
H C G T N V C R K P F S F K W
6
14351


648
T N V C R K P F S F K W W N M
6
14352


649
N V C R K P F S F K W W N M V
6
14353


650
V C R K P F S F K W W N M V P
6
14354


203
T S S Y D A K R A Q V F H I C
5
14355


376
L Q D F P Y K R I L C P D F G
5
14356


388
D F G Y V T R E P R D R S V S
5
14357


497
F K L F Q E K Q K C G H G R A
5
14358


131
R N F V D K R Q W V W G P S G
3
14359


529
N Q V L S N K D L I N Y N K F
3
14360


589
N M L V L G K H L G I P K P F
3
14361


2
S L Q R I V R V S L E H P T S
1
14362


63
A D T R R W R F D A T L E I I
1
14363


125
C E G R Q D R N F V D K R Q W
1
14364


192
A L F D D H K L V L H T S S Y
1
14365


204
S S Y D A K R A Q V F H I C G
1
14366


221
D V C E A Y R H V L G Q D K V
1
14367


235
V S Y E V P R L H G D E E R F
1
14368


308
V Y V C R V R N N T C F V D A
1
14369


322
A V A E L A R K A G C K L T I
1
14370


327
A R K A C C K L T I C P Q A E
1
14371


337
C P Q A E N R N D R W I Q D E
1
14372


356
Y V Q A P H K T L P V V F D S
1
14373


366
V V F D S P R N G E L Q D F P
1
14374


393
T R E P R D R S V S G L D S F
1
14375


434
L P G S S G R R V T Q V V R D
1
14376


441
R V T Q V V R D F L H A Q K V
1
14377


478
V P A P D G K G F R M L L A S
1
14378


481
P D C K G F R M L L A S P G A
1
14379


492
S P G A C F K L F Q E K Q K C
1
14380


504
Q K C G H G R A L L F Q G V V
1
14381


518
V D D E Q V K T I S I N Q V L
1
14382


550
W N R E V L K R E L G L A E C
1
14383


568
D I P Q L F K T E R K K A T A
1
14384


571
Q L F K T E R K K A T A F F P
1
14385


573
F K T E R K K A T A F F P D L
1
14386


595
K H L G I P K P F G P I I N G
1
14387


56
M E R G R E R A D T R R W R F
−5
14388


60
R E R A D T R R W R F D A T L
−5
14389


122
D L N C E G R Q D R N F V D K
−5
14390


150
L L V N C D R D D P S C D V Q
−5
14391


242
L H G D E E R F F V E G L S F
−5
14392


340
A E N R N D R W I Q D E M E L
−5
14393


377
Q D F P Y K R T L G P D F G Y
−5
14394


415
P V V A N G K E Y P L G R I L
−5
14395


421
K E Y P L G R I L I G G N L P
−5
14396


499
L F Q E K Q K C G H G R A L L
−5
14397


536
D L I N Y N K F V Q S C I D W
−5
14398


546
S C I D W N R E V L K R E L G
−5
14399


572
L F K T E R K K A T A F F P D
−5
14400


612
C L E E K V R S L L E P L G L
−5
14401


646
C G T N V C R K P F S F K W W
−5
14402







184P1E2 v.2: HLA Peptide Scoring Results DRB1 0401


15-mers SYFPE1THI










4
V A P W I M T P S T L A P L E
22
14403


2
F R V A P W I M T P S T L A P
18
14404


15
A P L E V Y V C R V R N N T C
18
14405


1
V F R V A P W I M T P S T L A
14
14406


5
A P W I M T P S T L A P L E V
14
14407


6
P W I M T P S T L A P L E V Y
14
14408


11
P S T L A P L E V Y V C R V R
14
14409


8
I M T P S T L A P L E V Y V C
12
14410


13
T L A P L E V Y V C R V R N N
12
14411


14
L A P L E V Y V C R V R N N T
8
14412


3
R V A P W I M T P S T L A P L
6
14413


9
M T P S T L A P L E V Y V C R
6
14414


10
T P S T L A P L E V Y V C R V
6
14415


12
S T L A P L E V Y V C R V R N
6
14416







184P1E2 v.3: HLA Peptide Scoring Results DRB1 0401


15-mers SYFPE1THI










3
V G H V D E F L S F V P V P D
20
14417


6
V D E F L S F V P V P D G K G
16
14418


9
F L S F V P V P D G K G F R M
16
14419


7
D E F L S F V P V P D G K G F
14
14420


12
F V P V P D G K G F R N L L A
14
14421


2
A V G H V D E F L S F V P V P
12
14422


4
G H V D E F L S F V P V P D G
12
14423


14
P V P D G K G F R M L L A S P
12
14424


15
V P D G K G F R M L L A S P G
12
14425


10
L S F V P V P D G K G F R M L
8
14426


1
L A V G H V D E F L S F V P V
6
14427


8
E F L S F V P V P D G K G F R
6
14428


11
S F V P V P D G K G F R M L L
6
14429


13
V P V P D G K G F R M L L A S
1
14430



















TABLE LI








SEQ.





ID


Pos
1 2 3 4 5 6 7 8 9 0 1 2 3 4 5
score
NO.















184P1E 2 v.1: HLA Peptide Scoring Results DRB1 0401


15-mers SYFPEITHI










587
L V N M L V L G K H L G I P K
28
14431


320
V D A V A E L A R K A G C K L
26
14432


386
G P D F G Y V T R E P R D R S
24
14433


389
F G Y V T R E P R D R S V S G
24
14434


439
G R R V T Q V V R D F L H A Q
24
14435


446
V R D F L H A Q K V Q P P V E
24
14436


462
F V D W L A V G H V D E F L S
24
14437


284
I F T D T V V F R V A P W I M
23
14438


404
L D S F G N L E V S P P V V A
22
14439


483
G K G F R M L L A S P G A C F
22
14440


549
D W N R E V L K R E L G L A E
22
14441


569
I P Q L F K T E R K K A T A F
22
14442


354
L G Y V Q A P H K T L P V V F
21
14443


5
R I V R V S L E H P T S A V C
20
14444


20
V A G V E T L V D I Y G S V P
20
14445


71
D A T L E I I V V N N S P S N
20
14446


148
G I L L V N C D R D D P S C D
20
14447


178
E D M S V M V L R T Q G P A A
20
14448


194
F D D H K L V L H T S S Y D A
20
14449


207
D A K R A Q V F H I C G P E D
20
14450


233
D K V S Y E V P R L H G D E E
20
14451


306
L E V Y V C R V R N N T C F V
20
14452


413
S P P V V A N G K E Y P L G R
20
14453


566
I I D I P Q L F K T E R K K A
20
14454


593
L G K H L G I P K P F G P I I
20
14455


23
V S T L V D I Y G S V P E C T
19
14456


36
G T E M F E V Y G T P G V D I
19
14457


73
T L S I I V V M N S P S N D L
19
14458


179
D M S V M V L R T Q G P A A L
19
14459


316
N T C F V D A V A E L A R K A
19
14460


465
W L A V G H V D E F L S F V P
19
14461


581
T A F F P D L V N M L V L G K
19
14462


614
E E K V R S L L E P L G L H C
19
14463


3
L Q R I V R V S L E H P T S A
18
14464


48
V D I Y I S P N M S R C R S R
18
14465


246
E S R F F V E G L S F P D A G
18
14466


248
R F F V S G L S F P D A G F T
18
14467


258
D A G F T G L I S F H V T L L
18
14468


353
E L G Y V Q A P H K T L P V V
18
14469


472
D S F L S F V P A P D G K G F
18
14470


497
F K L F Q E K Q K C G H G R A
18
14471


511
A L L F Q G V V D D E Q V K T
18
14472


570
P Q L F K T F R K K A T A F F
18
14473


631
I D D F T P Y H M L H G E V H
18
14474


93
Q I S Y H S S H E P L P L A Y
17
14475


104
P L A Y A V L Y L T C V D I S
17
14476


264
L I S F H V T L L D D S N E D
17
14477


293
V A P W I M T P S T L P P L E
17
14478


344
N D R W I Q D E M E L G Y V Q
17
14479


420
G K E Y P L G R I L I G G N L
17
14480


474
F L S F V P A P D G K G F R M
17
14481


516
G V V D D E Q V K T I S I N Q
17
14482


610
C C C L E E K V R S L L E P L
17
14483


634
F T P Y H M L H G E V H C G T
17
14484


27
V D I Y G S V P E G T E M F E
16
14485


50
I Y I S P N M E R G R E R A D
16
14486


52
I S P N M E R G R E R A D T R
16
14487


61
E R A D T R R W R F D A T L E
16
14488


136
K R Q W V W G P S G Y G G I L
16
14489


143
P S G Y G G I L L V N C D R D
16
14490


177
L E D M S V M V L R T Q G P A
16
14491


190
P A A L F D D H K L V L H T S
16
14492


223
C E A Y R H V L G Q D K V S Y
16
14493


282
S P I F T D T V V F R V A P W
16
14494


283
P I F T D T V V F R V A P W I
16
14495


304
P P L E V Y V C R V R N N T C
16
14496


375
E L Q D F P Y K R I L G P D F
16
14497


376
L Q D F P Y K R I L G P D F G
16
14498


378
D F P Y K R I L G P D F G Y V
16
14499


500
F Q E K Q K C G H G R A L L F
16
14500


571
Q L F K T E R K K A T A F F P
16
14501


600
P K P F C P I I N C C C C L E
16
14502


608
N G C C C L E E K V R S L L E
16
14503


637
Y H M L H G E V H C G T N V C
16
14504


644
V H C G T N V C R K P F S F K
16
14505


109
V L Y L T C V D I S L D C D L
15
14506


128
R Q D R N F V D K R Q W V W C
15
14507


219
P E D V C E A Y R H V L G Q D
15
14508


220
E D V C E A Y R H V L C Q D K
15
14509


364
L P V V F D S P R N G E L Q D
15
14510


391
Y V T R E P R D R S V S G L D
15
14511


419
N G K E Y P L G R I L I G G N
15
14512


456
Q P P V E L F V D W L A V G H
15
14513


458
P V E L F V D W L A V G H V D
15
14514


534
N K D L I N Y N K F V Q S C I
15
14515


548
I D W N R E V L K R E L G L A
15
14516


585
P D L V N M L V L G K H L G I
15
14517


645
H C G T N V C R K P F S F K W
15
14518


21
A G V E T L V D I Y G S V P E
14
14519


33
V P E G T E M F E V Y C T P G
14
14520


72
A T L E I I V V M N S P S N D
14
14521


89
D S H V Q I S Y H S S H E P L
14
14522


120
D C D L N C E C R Q D R N F V
14
14523


146
Y G G I L L V N C D R D D P S
14
14524


193
L F D D H K L V L H T S S Y D
14
14525


226
Y R H V L G Q D K V S Y E V P
14
14526


266
S F H V T L L D D S N E D F S
14
14527


287
D T V V F R V A P W I M T P S
14
14528


360
P H K T L P V V F D S P R N G
14
14529


407
F G N L E V S P P V V A N G K
14
14530


409
N L E V S P P V V A N G K E Y
14
14531


443
T Q V V R D F L H A Q K V Q P
14
14532


547
C I D W N R E V L K R E L G L
14
14533


588
V N M L V L G K H L G I P K P
14
14534


625
G L H C T F I D D F T P Y H M
14
14535


633
D F T P Y H M L H G E V H C G
14
14536


17
A V C V A G V E T L V D T Y G
13
14537


42
V Y G T P G V D I Y I S P N M
13
14538


74
L E I I V V M N S P S N D L N
13
14539


84
S N D L N D S H V Q I S Y H S
13
14540


107
Y A V L Y L T C V D I S L D C
13
14541


174
L Q D L E D M S V M V L R T Q
13
14542


210
R A Q V F H I C G P E D V C E
13
14543


229
V L G Q D K V S Y E V P R L H
13
14544


300
P S T L P P L E V Y V C R V R
13
14545


361
H K T L P V V F D S P R N G E
13
14546


363
T L P V V F D S P R N G E L Q
13
14547


424
P L G R I L I G G N L P G S S
13
14548


468
V G H V D E F L S F V P A P D
13
14549


482
D C K G F R M L L A S P C A C
13
14550


496
C F K L F Q E K Q K C G H G R
13
14551


522
Q V K T I S I N Q V L S N K D
13
14552


525
T I S I N Q V L S N K D L I N
13
14553


551
N R E V L K R E L G L A E C D
13
14554


589
N M L V L G K H L G I P K P F
13
14555


591
L V L G K H L G I P K P F G P
13
14556


6
I V R V S L E H P T S A V C V
12
14557


88
N D S H V Q I S Y H S S H F P
12
14558


91
H V Q I S Y H S S H E P L P L
12
14559


97
H S S H E P L P L A Y A V L Y
12
14560


106
A Y A V L Y L T C V D I S L D
12
14561


108
A V L Y L T C V D I S L D C D
12
14562


116
D I S L D C D L N C E G R Q D
12
14563


131
R N F V D K R Q W V W G P S G
12
14564


134
V D K R Q W V W G P S G Y G G
12
14565


137
R Q W V W G P S G Y G G I L L
12
14566


149
I L L V N C D R D D P S C D V
12
14567


168
D Q H V H C L Q D L E D N S V
12
14568


171
V H C L Q D L E D M S V M V L
12
14569


180
M S V M V L R T Q G P A A L F
12
14570


181
S V M V L R T Q G P A A L F D
12
14571


197
H K L V L H T S S Y D A K R A
12
14572


236
S Y E V P R L H G D E E R F F
12
14573


269
V T L L D D S N E D F S A S P
12
14574


290
V F R V A P W I M T P S T L P
12
14575


294
A P W I M T P S T L P P L S V
12
14576


307
E V Y V C R V R N N T C F V D
12
14577


317
T C F V D A V A E L A R K A G
12
14578


348
I Q D E M E L G Y V Q A P H K
12
14579


349
Q D E M E L G Y V Q A P H K T
12
14580


351
E N E L G Y V Q A P H K T L P
12
14581


379
F P Y K R I L G P D F G Y V T
12
14582


395
E P R D R S V S G L D S F G N
12
14583


398
D R S V S G L D S F G N L E V
12
14584


422
E Y P L G R I L I G G N L P G
12
14585


425
L G R I L I G G N L P G S S G
12
14586


426
G R I L I G G N L P G S S G R
12
14587


427
R I L I G G N L P G S S G R R
12
14588


428
I L I G G N L P G S S G R R V
12
14589


447
R D F L H A Q K V Q P P V E L
12
14590


460
E L F V D W L A V G H V D E F
12
14591


469
G H V D E F L S F V P A P D G
12
14592


471
V D E F L S F V P A P D G K G
12
14593


484
K G F R M L L A S P G A C F K
12
14594


520
D E Q V K T I S I N Q V L S N
12
14595


535
K D L I N Y N K F V Q S C I D
12
14596


537
L I N Y N K F V Q S C I D W N
12
14597


563
E C D I I D I P Q L F K T E R
12
14598


584
F P D L V N M L V L G K H L G
12
14599


594
G K H L G I P K P F G P I I N
12
14600


618
R S L L E P L G L H C T F I D
12
14601


636
P Y H M L H G E V H C G T N V
12
14602


37
T E M F E V Y G T P G V D I Y
11
14603


40
F E V Y G T P G V D I Y I S P
11
14604


67
R W R F D A T L E I I V V M N
11
14605


130
D R N F V D K R Q W V W G P S
11
14606


164
Q D N C D Q H V H C L Q D L E
11
14607


191
A A L F D D H K L V L H T S S
11
14608


203
T S S Y D A K R A Q V F H I C
11
14609


234
K V S Y E V P R L H G D E E R
11
14610


288
T V V F R V A P W I M T P S T
11
14611


303
L P P L E V Y V C R V R N N T
11
14612


313
V R N N T C F V D A V A E L A
11
14613


388
D F G Y V T R E P R D R S V S
11
14614


580
A T A F F P D L V N M L V L G
11
14615


11
L E H P T S A V C V A G V E T
10
14616


59
G R E R A D T R R W R F D A T
10
14617


65
T R R W R F D A T L S I I V V
10
14618


129
Q D R N F V D K R Q W V W G P
10
14619


138
Q W V W G P S G Y C G I L L V
10
14620


211
A Q V F H I C G P E D V C E A
10
14621


232
Q D K V S Y E V P R L H G D E
10
14622


247
E R F F V E G L S F P D A G F
10
14623


253
G L S F P D A G F T G L I S F
10
14624


276
N E D F S A S P I F T D T V V
10
14625


359
A P H K T L P V V F D S P R N
10
14626


365
P V V F D S P R N G E L Q D F
10
14627


374
G E L Q D F P Y K R I L G P D
10
14628


385
L G P D F G Y V T R E P R D R
10
14629


432
G N L P G S S G R S V T Q V V
10
14630


459
V E L F V D W L A V C H V D E
10
14631


494
G A C F K L F Q E K Q K C G H
10
14632


540
Y N K F V Q S C I D W N R E V
10
14633


627
H C T F I D D F T P Y H M L H
10
14634


630
F I D D F T P Y H M L H G E V
10
14635


643
E V H C G T N V C R K P F S F
10
14636


58
R G R E R A D T R R W R F D A
9
14637


102
P L P L A Y A V L Y L T C V D
9
14638


189
G P A A L F D D H K L V L H T
9
14639


201
L H T S S Y D A K R A Q V F H
9
14640


244
G D E E R F F V E G L S F P D
9
14641


262
T G L I S F H V T L L D D S N
9
14642


338
P Q A E N R N D R W I Q D E M
9
14643


431
G G N L P G S S G R R V T Q V
9
14644


479
P A P D G K G F R M L L A S P
9
14645


495
A C F K L F Q E K Q K C G H G
9
14646


502
E K Q K C G H G R A L L F Q G
9
14647


510
R A L L F Q G V V D D E Q V K
9
14648


527
S I N Q V L S N K D L T N Y N
9
14649


544
V Q S C I D W N R E V L K R E
9
14650


26
L V D I Y G S V P E G T E M F
8
14651


35
E G T E M F E V Y G T P G V D
8
14652


54
P N M E R G R E R A D T R R W
8
14653


83
P S N D L N D S H V Q I S Y H
8
14654


92
V Q I S Y H S S H E P L P L A
8
14655


123
L N C E G R Q D R N F V D K R
8
14656


133
F V D K R Q W V W G P S G Y G
8
14657


145
G Y G G I L L V N C D R D D P
8
14658


147
G G I L L V N C D R D D P S C
8
14659


162
D V Q D N C D Q H V H C L Q D
8
14660


176
D L E D M S V M V L R T Q G P
8
14661


200
V L H T S S Y D A K R A Q V F
8
14662


202
H T S S Y D A K R A Q V F H I
8
14663


224
E A Y R H V L G Q D K V S Y E
8
14664


235
V S Y E V P R L H G D E E R F
8
14665


240
P R L H G D E E R F F V E G L
8
14666


260
G F T G L I S F H V T L L D D
8
14667


286
T D T V V F R V A P W I M T P
8
14668


295
P W I M T P S T L P P L E V Y
8
14669


302
T L P P L E V Y V C R V R N N
8
14670


305
P L E V Y V C R V R N N T C F
8
14671


319
F V D A V A E L A R K A G C K
8
14672


321
D A V A E L A R K A G C K L T
8
14673


325
E L A R K A G C K L T I C P Q
8
14674


331
G C K L T I C P Q A E N R N D
8
14675


333
K L T I C P Q A E N R N D R W
8
14676


335
T I C P Q A E N R N D R W I Q
8
14677


373
N G E L Q D F P Y K R I L G P
8
14678


410
L E V S P P v V A N C K E Y P
8
14679


411
E V S P P V V A N G K E Y P L
8
14680


433
N L P G S S G R R V T Q V V R
8
14681


440
R R V T Q V V R D F L H A Q K
8
14682


452
A Q K V Q P P V E L F V D W L
8
14683


476
S F V P A P D G K G F R M L L
8
14684


490
L A S P G A C F K L F Q E K Q
8
14685


493
P G A C F K L F Q E K Q K C G
8
14686


542
K F V Q S C I D W N R E V L K
8
14687


583
F F P D L V N M L V L G K H L
8
14688


619
S L L E P L C L H C T F I D D
8
14689


650
V C R K P F S F K W W N M V P
8
14690


1
M S L Q R I V R V S L E H P T
7
14691


4
Q R I V R V S L E H P T S A V
7
14692


8
R V S L E H P T S A V C V A G
7
14693


10
S L E H P T S A V C V A G V E
7
14694


15
T S A V C V A G V E T L V D I
7
14695


43
Y G T P G V D I Y I S P N M E
7
14696


49
D I Y I S P N M E R G R E R A
7
14697


53
S P N M E R G R E R A D T R R
7
14698


68
W R F D A T L E I I V V M N S
7
14699


77
I V V M N S P S N D L N D S H
7
14700


87
L N D S H V Q I S Y H S S H E
7
14701


95
S Y H S S H E P L P L A Y A V
7
14702


99
S H E P L P L A Y A V L Y L T
7
14703


100
H E P L P L A Y A V L Y L T C
7
14704


113
T C V D I S L D C D L N C E G
7
14705


195
D D H K L V L H T S S Y D A K
7
14706


208
A K R A Q V F H I C G P E D V
7
14707


225
A Y R H V L G Q D K V S Y E V
7
14708


239
V P R L H G D E E R F F V E G
7
14709


245
D E E R F F V E G L S F P D A
7
14710


252
E G L S F P D A G F T C L I S
7
14711


255
S F P D A G F T G L I S F H V
7
14712


259
A C F T C L I S F H V T L L D
7
14713


261
F T G L I S F H V T L L D D S
7
14714


263
G L I S F H V T L L D D S N E
7
14715


281
A S P I F T D T V V F R V A P
7
14716


310
V C R V R N N T C F V D A V A
7
14717


323
V A E L A R K A G C K L T I C
7
14718


382
K R I L G P D F G Y V T R E P
7
14719


402
S G L D S F G N L E V S P P V
7
14720


403
Q L D S F G N L E V S P P V V
7
14721


406
S F G N L E V S P P V V A N G
7
14722


415
P V V A N G K E Y P L C R I L
7
14723


436
G S S G R R V T Q V V R D F L
7
14724


441
R V T Q V V R D F L H A Q K V
7
14725


444
Q V V R D F L H A Q K V Q P P
7
14726


449
F L H A Q K V Q P P V E L F V
7
14727


453
Q K V Q P P V E L F V D W L A
7
14728


463
V D W L A V G H V D E F L S F
7
14729


470
H V D E F L S F V P A P D G K
7
14730


480
A P D G K G F R M L L A S P G
7
14731


507
G H G R A L L F Q C V V D D E
7
14732


512
L L F Q G V V D D E Q V K T I
7
14733


514
F Q G V V D D E Q V K T I S I
7
14734


526
I S I N Q V L S N K D L I N Y
7
14735


528
I N Q V L S N K D L I N Y N K
7
14736


532
L S N K D L I N Y N K F V Q S
7
14737


538
I N Y N K F V Q S C I D W N K
7
14738


545
Q S C I D W N R E V B K R E L
7
14739


556
K R E L G L A E C D I I D I P
7
14740


559
L G L A E C D I I D I P Q L F
7
14741


564
C D I I D I P Q L F K T E R K
7
14742


582
A F F P D L V N M L V L G K H
7
14743


596
H L G I P K P F G P I I N G C
7
14744


603
F G P I I N G C C C L E E K V
7
14745


607
I N G C C C L E E K V R S L L
7
14746


611
C C L E E K V R S L L E P L G
7
14747


612
C L E E K V R S L L E P L G L
7
14748


621
L E P L G L H C T F I D D F T
7
14749


12
E H P T S A V C V A G V E T L
6
14750


14
P T S A V C V A G V E T L V D
6
14751


24
E T L V D I Y G S V P E G T E
6
14752


25
T L V D I Y G S V P E G T E N
6
14753


30
Y G S V P E G T E M F E V Y G
6
14754


39
M F E V Y G T P G V D I Y I S
6
14755


44
G T P G V D I Y I S P N M E R
6
14756


45
T P G V D I Y I S P N M E R G
6
14757


46
P G V D I Y I S P N M E R G R
6
14758


47
G V D I Y I S P N M E R G R E
6
14759


70
F D A T L E I I V V M N S P S
6
14760


75
E I I V V M N S P S N D L N D
6
14761


76
I I V V M N S P S N D L N D S
6
14762


79
V M N S P S N D L N D S H V Q
6
14763


81
N S P S N D L N D S H V Q I S
6
14764


86
D L N D S H V Q I S Y H S S H
6
14765


103
L P L A Y A V L Y L T C V D I
6
14766


111
Y L T C V D I S L D C D L N C
6
14767


112
L T C V D I S L D C D L N C E
6
14768


114
C V D I S L D C D L N C E G R
6
14769


117
I S L D C D L N C E G R Q D R
6
14770


132
N F V D K R Q W V W G P S G Y
6
14771


144
S G Y G C I L L V N C D R D D
6
14772


157
D D P S C D V Q D N C D Q H V
6
14773


160
S C D V Q D N C D Q H V H C L
6
14774


165
D N C D Q H V H C L Q D L E D
6
14775


170
H V H C L Q D L E D M S V M V
6
14776


182
V M V L R T Q G P A A L F D D
6
14777


187
T Q G P A A L F D D H K L V L
6
14778


196
D H K L V L H T S S Y D A K R
6
14779


198
K L V L H T S S Y D A K K A Q
6
14780


213
V F H I C G P E D V C E A Y R
6
14781


214
F H I C G P E D V C E A Y R H
6
14782


216
I C G P E D V C E A Y R H V L
6
14783


227
R H V L G Q D K V S Y E V P R
6
14784


242
L H G D E E R F F V E C L S F
6
14785


249
F F V E G L S F P D A G F T G
6
14786


251
V E G L S F P D A G F T G L I
6
14787


265
I S F H V T L L D D S N F D F
6
14788


268
H V T L L D D S N E D F S A S
6
14789


270
T L L D D S N E D F S A S P I
6
14790


271
L L D D S N F D F S A S P I F
6
14791


272
L D D S N E D F S A S P I F T
6
14792


278
D F S A S P I F T D T V V F R
6
14793


289
V V F R V A P W I N T P S T L
6
14794


291
F R V A P W I M T P S T L P P
6
14795


292
R V A P W I M T P S T L P P L
6
14796


297
I M T P S T L P P L E V Y V C
6
14797


311
C R V R N N T C F V D A V A E
6
14798


314
R N N T C F V D A V A E L A R
6
14799


327
A R K A G C K L T I C P Q A E
6
14800


328
R K A G C K L T I C P Q A E N
6
14801


329
K A G C K L T I C P Q A E N R
6
14802


330
A G C K L T I C P Q A E N R N
6
14803


342
N R N D R W I Q D E M E L G Y
6
14804


345
D R W I Q D E M E L G Y V Q A
6
14805


346
R W I Q D E M E L G Y V Q A P
6
14806


358
Q A P H K T L P V V F D S P R
6
14807


369
D S P R N G E L Q D F P Y K R
6
14808


370
S P R N G E L Q D F P Y K R I
6
14809


381
Y K R I L G P D F G Y V T R E
6
14810


392
V T R E P R D R S V S G L D S
6
14811


397
R D R S V S G L D S F G N L E
6
14812


401
V S G L D S F G N L E V S P P
6
14813


414
P P V V A N G K E Y P L G R I
6
14814


421
K E Y P L G R I L I G G N L P
6
14815


423
Y P L G R I L I G G N L P G S
6
14816


442
V T Q V V R D F L H A Q K V Q
6
14817


455
V Q P P V E L F V D W L A V G
6
14818


457
P P V E L F V D W L A V G H V
6
14819


475
L S F V P A P D G K G F R M L
6
14820


481
P D C K G F K N L L A S P C A
6
14821


485
G F R M L L A S P G A C F K L
6
14822


486
F R M L L A S P G A C F K L F
6
14823


487
R M L L A S P G A C F K L F Q
6
14824


498
K L F Q E K Q K C G H G R A L
6
14825


505
K C G H G R A L L F Q G V V D
6
14826


506
C G H G R A L L F Q G V V D D
6
14827


509
G R A L L F Q G V V D D E Q V
6
14828


515
Q C V V D D E Q V K T I S I N
6
14829


517
V V D D E Q V K T I S I N Q V
6
14830


523
V K T I S I N Q V L S N K D L
6
14831


529
N Q V L S N K D L I N Y N K F
6
14832


531
V L S N K D L I N Y N K F V Q
6
14833


541
N K F V Q S C I D W N R K V L
6
14834


552
R E V L K R E L G L A E C D I
6
14835


553
E V L K R E L G L A E C D I I
6
14836


555
L K R E L G L A E C D I I D I
6
14837


558
E L G L A E C D I I D I P Q L
6
14838


560
G L A E C D I I D I P Q L F K
6
14839


561
L A E C D I I D I P Q L F K T
6
14840


574
K T E R K K A T A F F P D L V
6
14841


586
D L V N M L V L G K H L C I P
6
14842


590
M L V L G K H L G I P K P F G
6
14843


598
G I P K P F C P I I N C C C C
6
14844


601
K P F G P I I N G C C C L E E
6
14845


604
G P I I N G C C C L E E K V R
6
14846


615
E K V R S L L E P L G L H C T
6
14847


617
V R S L L E P L G L H C T F I
6
14848


620
L L E P L G L H C T F I D D F
6
14849


623
P L G L H C T F I D D F T P Y
6
14850


628
C T F I D D F T P Y H M L H G
6
14851


638
H M L H G E V H C G T N V C R
6
14852


641
H G F V H C G T N V C R K P F
6
14853


647
G T N V C R K P F S F K W W N
6
14854


13
H P T S A V C V A G V E T L V
4
14855


41
E V Y G T P C V D I Y I S P N
4
14856


127
G R Q D R N F V D K R Q W V W
4
14857


2
S L Q R I V R V S L E H P T S
3
14858


16
S A V C V A G V E T L V D I Y
3
14859


57
E R G R E R A D T R R W R F D
3
14860


69
R F D A T L E I I V V M N S P
3
14861


175
Q D L E D M S V M V L R T Q G
3
14862


183
M V L R T Q G P A A L F D D H
3
14863


199
L V L H T S S Y D A K R A Q V
3
14864


206
Y D A K R A Q V F H I C G P E
3
14865


217
C G P E D V C E A Y R H V L G
3
14866


298
M T P S T L P P L E V Y V C R
3
14867


318
C F V D A V A E L A R K A G C
3
14868


362
K T L P V V F D S P R N G E L
3
14869


387
P D F G Y V T R E P R D R S V
3
14870


394
R E P R D R S V S G L D S F G
3
14871


435
P G S S G R R V T Q V V R D F
3
14872


437
S S G R R V T Q V V R D F L H
3
14873


438
S G R R V T Q V V R D F L H A
3
14874


461
L F V D W L A V G H V D E F L
3
14875


521
E Q V K T I S I N Q V L S N K
3
14876


577
R K K A T A F F P D L V N M L
3
14877


56
M E R G R E R A D T R R W R F
2
14878


66
R R W R F D A T L E I I V V M
2
14879


85
N D L N D S H V Q I S Y H S S
2
14880


118
S L D C D L N C E G R Q D R N
2
14881


121
C D L N C E G R Q D R N F V D
2
14882


156
R D D P S C D V Q D N C D Q H
2
14883


215
H I C G P E D V C E A Y R H V
2
14884


222
V C F A Y R H V L G Q D K V S
2
14885


228
H V L G Q D K V S Y E V P R L
2
14886


231
G Q D K V S Y E V P R L H G D
2
14887


238
E V P R L H G D E E R F F V E
2
14888


301
S T L P P L E V Y V C R V R N
2
14889


315
N N T C F V D A V A E L A R K
2
14890


336
I C P Q A E N R N D R W I Q D
2
14891


350
D E M E L G Y V Q A P H K T L
2
14892


352
M E L G Y V Q A P H K T L P V
2
14893


384
I L G P D F G Y V T R E P R D
2
14894


405
D S F G N L E V S P P V V A N
2
14895


417
V A N G K E Y P L G R I L I G
2
14896


4l8
A N G K E Y P L G R I L I G C
2
14897


430
I G G N L P G S S C R R V T Q
2
14898


445
V V R D F L H A Q K V Q P P V
2
14899


448
D F L H A Q K V Q P P V E L F
2
14900


464
D W L A V G H V D E F L S F V
2
14901


473
E F L S F V P A P D C K G F R
2
14902


477
F V P A P D G K G F R M L L A
2
14903


513
L F Q G V V D D E Q V K T I S
2
14904


518
V D D E Q V K T I S I N Q V L
2
14905


524
K T I S I N Q V L S N K D L I
2
14906


539
N Y N K F V Q S C I D W N R E
2
14907


567
I D I P Q L F K T E R K K A T
2
14908


568
D I P Q L F K T E R K K A T A
2
14909


642
G E V H C G T N V C R K P F S
2
14910


18
V C V A G V E T L V D I Y G S
1
14911


19
C V A G V E T L V D I Y G S V
1
14912


22
G V E T L V D I Y G S V P E G
1
14913


28
D I Y G S V P E G T E M F E V
1
14914


32
S V P E G T E M F E V Y G T P
1
14915


38
E M F E V Y G T P G V D I Y I
1
14916


si
Y I S P N M E R G R E R A D T
1
14917


60
R E R A D T R R W R F D A T L
1
14918


64
D T R R W R F D A T L E I I V
1
14919


82
S P S N D L N D S H V Q I S Y
1
14920


105
L A Y A V L Y L T C V D I S L
1
14921


110
L Y L T C V D I S L D C D L N
1
14922


119
L D C D L N C E G R Q D R N F
1
14923


122
D L N C E G R Q D R N F V D K
1
14924


124
N C E G R Q D R N F V D K R Q
1
14925


126
E G R Q D R N F V D K R Q W V
1
14926


135
D K R Q W V W G P S G Y G G I
1
14927


139
W V W G P S G Y G G I L L V N
1
14928


140
V W G P S G Y G G I L L V N C
1
14929


141
W G P S G Y G G I L L V N C D
1
14930


142
G P S G Y G G I L L V N C D R
1
14931


153
N C D R D D P S C D V Q D N C
1
14932


158
D P S C D V Q D N C D Q H V H
1
14933


161
C D V Q D N C D Q H V H C L Q
1
14934


166
N C D Q H V H C L Q D L E D M
1
14935


167
C D Q H V H C L Q D L E D M S
1
14936


172
H C L Q D L E D M S V M V L R
1
14937


188
Q G P A A L F D D H K L V L H
1
14938


212
Q V F H I C G P E D V C E A Y
1
14939


218
G P E D V C E A Y R H V L G Q
1
14940


230
L G Q D K V S Y E V P R L H G
1
14941


241
R L H G D E S R F F V E G L S
1
14942


254
L S F P D A G F T G L I S F H
1
14943


257
P D A G F T G L I S F H V T L
1
14944


274
D S N E D F S A S P I F T D T
1
14945


275
S N E D F S A S P I F T D T V
1
14946


279
F S A S P I F T D T V V F R V
1
14947


280
S A S P I F T D T V V F R V A
1
14948


285
F T D T V V F R V A P W I M T
1
14949


296
W I M T P S T L P P L S V Y V
1
14950


308
V Y V C R V R N N T C F V D A
1
14951


324
A E L A R K A G C K L T I C P
1
14952


326
L A R K A G C K L T I C P Q A
1
14953


334
L T I C P Q A E N R N D R W I
1
14954


337
C P Q A E N R N D R W I Q D E
1
14955


347
W I Q D E H E L G Y V Q A P H
1
14956


356
Y V Q A P H K T L P V V F D S
1
14957


357
V Q A P H K T L P V V F D S P
1
14958


366
V V F D S P R N G E L Q D F P
1
14959


372
R N G E L Q D F P Y K R I L G
1
14960


390
G Y V T R E P R D R S V S G L
1
14961


396
P R D R S V S G L D S F G N L
1
14962


400
S V S G L D S F G N L E V S P
1
14963


412
V S P P V V A N G K E Y P L G
1
14964


450
L H A Q K V Q P P V E L F V D
1
14965


451
H A Q K V Q P P V E L F V D W
1
14966


454
K V Q P P V E L F V D W L A V
1
14967


466
L A V G H V D E F L S F V P A
1
14968


478
V P A P D G K G F R M L L A S
1
14969


488
M L L A S P G A C F K L F Q E
1
14970


489
L L A S P G A C F K L F Q E K
1
14971


501
Q E K Q K C G H G R A L L F Q
1
14972


503
K Q K C G H G R A L L F Q G V
1
14973


508
H G R A L L F Q G V V D D E Q
1
14974


543
F V Q S C I D W N R E V L K R
1
14975


546
S C I D W N R E V L K R E L G
1
14976


557
R E L G L A E C D I I D I P Q
1
14977


562
A E C D I I D I P Q L F K T E
1
14978


578
K K A T A F F P D L V N M L V
1
14979


579
K A T A F F P D L V N M L V L
1
14980


597
L G I P K P F G P I I N G C C
1
14981


599
I P K P F G P I I N G C C C L
1
14982


606
I I N G C C C L E E K V R S L
1
14983


609
G C C C L E E K V R S L L E P
1
14984


616
K V R S L L E P L G L H C T F
1
14985


626
L H C T F I D D F T P Y H M L
1
14986


639
M L H G E V H C G T N V C R K
1
14987


640
L H G E V H C G T N V C R K P
1
14988


648
T N V C R K P F S F K W W N M
1
14989







184P1E2 v.2: HLA Peptide Scoring Results DRB1 1101


15-mers SYF PE ITIII










4
V A P W I M T P S T L A P L E
17
14990


15
A P L E V Y V C R V R N N T C
16
14991


11
P S T L A P L E V Y V C R V R
13
14992


1
V F R V A P W I M T P S T L A
12
14993


5
A P W I M T P S T L A P L E V
12
14994


14
L A P L E V Y V C R V R N N T
11
14995


6
P W I M T P S T L A P L E V Y
8
14996


13
T L A P L E V Y V C R V R N N
8
14997


2
F R V A P W I M T P S T L A P
6
14998


3
R V A P W I M T P S T L A P L
6
14999


5
I M T P S T L A P L E V Y V C
6
15000


9
M T P S T L A P L F V Y V C R
3
15001


12
S T L A P L E V Y V C R V R N
2
15002


7
W I M T P S T L A P L E V Y V
1
15003







184P1E2 v.3: HLA Peptide Scoring Results DRB1 1101


15-mers SYFPEITHI










9
F L S F V P V P D G K G F R M
23
15004


7
D E F L S F V P V P D G K G F
18
15005


3
V G H V D E F L S F V P V P D
13
15006


6
V D E F L S F V P V P D G K G
12
15007


14
P V P D G K G F R M L L A S P
9
15008


5
H V D E F L S F V P V P D G K
8
15009


11
S F V P V P D G K G F R M L L
8
15010


12
F V P V P D G K G F R M L L A
8
15011


10
L S F V P V P D G K G F R M L
7
15012


15
V P D G K G F R M L L A S P G
7
15013


4
G H V D E F L S F V P V P D G
6
15014


8
E F L S F V P V P D G K G F R
4
15015


1
L A V G H V D E F L S F V P V
1
15016


13
V P V P D G K G F R M L L A S
1
15017
















TABLE LII





Search Peptides
















184P1E2 v.1:



For all 184P1E2 v.1 - nonamers. decamers and 15-mers


  1 MSLQRIVRVS LEHPTSAVCV AGVETLVDIY GSVPEGTEMF EVYGTPGVDI YISPNMERGR


 61 ERADTRRWRF DATLEIIVVM NSPSNDLNDS HVQISYHSSH EPLPLAYAVL YLTCVDISLD


121 CDLNCEGRQD RNFVDKRQWV WGPSGYGGIL LVNCDRDDPS CDVQDNCDQH VHCLQDLEDM


181 SVMVLRTQGP AALFDDHKLV LHTSSYDAKR AQVFHICGPE DVCEAYRHVL GQDKVSYEVP


241 RLHGDEERFF VEGLSFPDAG FTGLISFHVT LLDDSNEDFS ASPIFTDTVV FRVAPWIMTP


301 STLPPLEVYV CRVRNNTCFV DAVAELARKA GCKLTICPQA ENRNDRWIQD EMELGYVQAP


361 HKTLPVVFDS PRNGELQDFP YKRILGPDFG YVTREPRDRS VSGLDSFGNL EVSPPVVANG


421 KEYPLGRILI GGNLPGSSGR RVTQVVRDFL HAQKVQPPVE LFVDWLAVGH VDEFLSFVPA


481 PDGKGFRMLL ASPGACFKLF QEKQKCGHGR ALLFQGVVDD EQVKTISINQ VLSNKDLINY


541 NKFVQSCIDW NREVLKRELG LAECDIIDIP QLFKTERKKA TAFFPDLVNM LVLGKHLGIP


601 KPFGPIINGC CCLEEKVRSL LEPLGLHCTF IDDFTPYHML HGEVHCGTNV CRKPFSFKWW


661 NMVP (aa. 1-664)


184P1E2 v.2:


184P1E2 v.2 nonamers


WIMTPSTLAPLEVYVCR (aa. 296-312)


184P1E2 v.2 decarners


PWIMTPSTLAPLEVYVCRV (aa. 295-313)


184P1E2 v.2 1 5-mers


VFRVAPWIMTPSTLAPLEVYVCRVRNNTC (aa. 290-318)


184P1E2 v.3:


184P1F2 v.3 nonamers


DEFLSFVPVPDGKGFRM (aa. 472-488)


184P1E2 v.3 decamers


VDEFLSFVPVPDGKGFRML (aa. 471-489)


184P1E2 v.3 15-mers


LAVGHVDEFLSFVPVPDGKGFRMLLASPG (aa. 466-494)
















TABLE LIII





Clustal Alignment of the three 184P1E2 variants Showing SNP Modifications.
















v.1
MSLQRIVRVSLEHPTSAVCVAGVETLVDIYGSVPEGTEMFEVYGTPGVDIYISPNMERGR


v.2
MSLQRIVRVSLEHPTSAVCVAGVETLVDIYGSVPEGTEMFEVYGTPGVDIYISPNMERGR


v.3
MSLQRIVRVSLEHPTSAVCVAGVETLVDIYGSVPEGTEMFEVYGTPGVDIYISPNMERGR



************************************************************


 


v.1
ERADTRRWRFDATLEIIVVMNSPSNDLNDSHVQISYHSSHEPLPLAYAVLYLTCVDISLD


v.2
ERADTRRWRFDATLEIIVVMNSPSNDLNDSHVQISYHSSHEPLPLAYAVLYLTCVDISLD


v.3
ERADTRRWRFDATLEIIVVMNSPSNDLNDSHVQISYHSSHEPLPLAYAVLYLTCVDISLD



************************************************************


 


v.1
CDLNCEGRQDRNFVDKRQWVWGPSGYGGILLVNCDRDDPSCDVQDNCDQHVHCLQDLEDM


v.2
CDLNCEGRQDRNFVDKRQWVWGPSGYGGILLVNCDRDDPSCDVQDNCDQHVHCLQDLEDM


v.3
CDLNCEGRQDRNFVDKRQWVWGPSGYGGILLVNCDRDDPSCDVQDNCDQHVHCLQDLEDM



************************************************************


 


v.1
SVMVLRTQGPAALFDDHKLVLHTSSYDAKRAQVFHICGPEDVCEAYRHVLGQDKVSYEVP


v.2
SVMVLRTQGPAALFDDHKLVLHTSSYDAKRAQVFHICGPEDVCEAYRHVLGQDKVSYEVP


v.3
SVMVLRTQGPAALFDDHKLVLHTSSYDAKRAQVFHICGPEDVCEAYRHVLGQDKVSYEVP



************************************************************


 


v.1
RLHGDEERFFVEGLSFPDAGFTCLISFHVTLLDDSNEDFSASPIFTDTVVFRVAPWIMTP


v.2
RLHGDEERFFVEGLSFPDAGFTCLISFHVTLLDDSNEDFSASPIFTDTVVFRVAPWIMTP


v.3
RLHGDEERFFVEGLSFPDAGFTCLISFHVTLLDDSNEDFSASPIFTDTVVFRVAPWIMTP



************************************************************


 


v.1
STLPPLEVYVCRVRNNTCFVDAVAELARKAGCKLTICPQAENRNDRWIQDEMELGYVQAP


v.2
STLAPLEVYVCRVRNNTCFVDAVAELARKAGCKLTICPQAENRNDRWIQDEMELGYVQAP


v.3
STLPPLEVYVCRVRNNTCFVDAVAELARKAGCKLTICPQAENRNDRWIQDEMELGYVQAP



***.********************************************************


 


v.1
HKTLPVVFDSPRNGELQDFPYKRILGPDFCYVTREPRDRSVSGLDSFGNLEVSPPVVANG


v.2
HKTLPVVFDSPRNGELQDFPYKRILGPDFCYVTREPRDRSVSGLDSFGNLEVSPPVVANG


v.3
HKTLPVVFDSPRNGELQDFPYKRILGPDFCYVTREPRDRSVSGLDSFGNLEVSPPVVANG



************************************************************


 


v.1
KEYPLGRILIGGNLPGSSGRRVTQVVRDFLHAQKVQPPVELFVDWLAVGHVDEFLSFVPA


v.2
KEYPLGRILIGGNLPGSSGRRVTQVVRDFLHAQKVQPPVELFVDWLAVGHVDEFLSFVPA


v.3
KEYPLGRILIGGNLPGSSGRRVTQVVRDFLHAQKVQPPVELFVDWLAVGHVDEFLSFVPV



***********************************************************.


 


v.1
PDGKGFRMLLASPGACFKLFQEKQKCGHGRALLFQGVVDDEQVKTISINQVLSNKDLINY


v.2
PDGKGFRMLLASPGACFKLFQEKQKCGHGRALLFQGVVDDEQVKTISINQVLSNKDLINY


v.3
PDGKGFRMLLASPGACFKLFQEKQKCGHGRALLFQGVVDDEQVKTISINQVLSNKDLINY



************************************************************


 


v.1
NKFVQSCIDWNREVLKRELGLAECDIIDIPQLFKTERKKATAFFPDLVNMLVLGKHLGIP


v.2
NKFVQSCIDWNREVLKRELGLAECDIIDIPQLFKTERKKATAFFPDLVNMLVLGKHLGIP


v.3
NKFVQSCIDWNREVLKRELGLAECDIIDIPQLFKTERKKATAFFPDLVNMLVLGKHLGIP



************************************************************


 


v.1
KPFGPIINGCCCLEEKVRSLLEPLGLHCTFIDDFTPYHMLHGEVHCGTNVCRKPFSFKWW


v.2
KPFGPIINGCCCLEEKVRSLLEPLGLHCTFIDDFTPYHMLHGEVHCGTNVCRKPFSFKWW


v.3
KPFGPIINGCCCLEEKVRSLLEPLGLHCTFIDDFTPYHMLHGEVHCGTNVCRKPFSFKWW



************************************************************


 


v.1
NMVP


v.2
NMVP


v.3
NMVP



****

## Claims

1. A method for detecting the presence of a protein in a biological sample comprising the steps of:
contacting the biological sample with an antibody or fragment thereof which specifically binds to the protein, wherein the protein comprises the amino acid sequence of SEQ ID NO:5 or 7 and,
determining the presence or absence of a complex of the antibody or fragment thereof and the protein in the biological sample compared to a corresponding normal control sample.

2. The method of claim 1, wherein the antibody comprises a polyclonal antibody.

3. The method of claim 1, wherein the antibody comprises a monoclonal antibody.

4. The method of claim 1, wherein the sample is taken from a patient who has or who is suspected of having cancer.

5. The method of claim 4 wherein the cancer is a bladder, kidney, or lung cancer.