Abstract:
A purified preparation of a peptide consisting essentially of an amino acid sequence identical to that of a segment of a naturally-occurring human protein, said segment being of 10 to 30 residues in length, inclusive, wherein said peptide binds to a human major histocompatibility complex (MHC) class II allotype.

Description:
This application is a continuation-in-part of U.S. Ser. No. 07/925,460, filed Aug. 11, 1992, now abandoned. 
    
    
     The invention was made in the course of research funded in part by the U.S. Government under NIH Grant 5R35-CA47554; the U.S. Government therefore has certain rights in the invention. 
    
    
     The field of the invention is major histocompatibility complex (MHC) antigens. 
     BACKGROUND OF THE INVENTION 
     Major histocompatibility complex (MHC) class II antigens are cell surface receptors that orchestrate all specific immune responses in vertebrates. Humans possess three distinct MHC class II isotypes: DR, for which approximately 70 different allotypes are known; DQ, for which 33 different allotypes are known; and DP, for which 47 different allotypes are known. Each individual bears two to four DR alleles, two DQ alleles, and two DP alleles. 
     MHC receptors (both class I and class II) participate in the obligate first step of immune recognition by binding small protein fragments (peptides) derived from pathogens or other non-host sources, and presenting these peptides to the regulatory cells (T cells) of the immune system. In the absence of MHC presentation, T cells are incapable of recognizing pathogenic material. Cells that express MHC class II receptors are termed antigen presenting cells (APC). APCs ingest pathogenic organisms and other foreign materials by enveloping them in endosomic vesicles, then subjecting them to enzymatic and chemical degradation. Foreign proteins which are ingested by APCs are partially degraded or “processed” to yield a mixture of peptides, some of which are bound by MHC class II molecules that are en route to the surface. Once on the cell surface, MHC-bound peptides are available for T cell recognition. 
     MHC class II antigens are expressed on the surface of APCs as a trimolecular complex composed of an α chain, a p chain, and a processed peptide. Like most polypeptides that are expressed on the cell surface, both α and β chains contain short signal sequences at their NH 2  termini which target them to the endoplasmic reticulum (ER). Within the ER the class II α/β chain complex associates with an lo additional protein termed the invariant chain (Ii). Association with Ii is proposed to block the premature acquisition of peptides (by blocking the peptide binding cleft of the MHC heterodimer), promote stable α/β interaction, and direct subsequent intracellular trafficking of the complex to endosomal vesicles. In the endosomes, Ii is removed by a process involving proteolysis; this exposes the peptide binding cleft, thus allowing peptides present in the endosome to bind to the MHC molecule. The class II/peptide complex is transported from the endosomes to the cell surface where it becomes accessible to T-cell recognition and subsequent activation of immune responses. Class II MHC molecules bind not only to peptides derived from exogenous (ingested) proteins, but also to those produced by degradation of endogenous (self) proteins. The amount of each species of peptide which binds class II is determined by its local concentration and its relative binding affinity for the given class II binding groove, with the various allotypes displaying different peptide-binding specificities. 
     Early during fetal development, the mammalian immune system is “tolerized”, or taught not to react, to self-peptides. The stability and maintenance of this system is critical for ensuring that an animal does not generate an immune response against self. A breakdown of this system gives rise to autoimmune conditions such as diabetes, rheumatoid arthritis and multiple sclerosis. Current technologies intended to manipulate the immune system into reestablishing proper nonresponsiveness include protocols involving the intravenous delivery of synthetic, high affinity binding peptides as blocking peptides. 
     Vaccination can generate protective immunity against a pathogenic organism by stimulating an antibody-mediated and/or a T cell-mediated response. Most of the current vaccination strategies still use relatively crude preparations, such as attenuated or inactivated viruses. These vaccines often generate both antibody- and cell-mediated immunity, and do not allow one to modulate the type of immune response generated. Moreover, in many diseases the generation of the wrong type of response can result in an exacerbated disease state. 
     SUMMARY OF THE INVENTION 
     In the work disclosed herein, naturally processed peptides bound to six of the some 70 known human MHC class II DR allotypes (HLA-DR1, HLA-DR2, HLA-DR3, HLA-DR4, HLA-DR7, and HLA-DR8) have been characterized. These peptides were found to be predominantly derived from self proteins rather than foreign proteins. Several self peptide families have been identified with the unexpected property of degenerate binding: that is, a given self-peptide will bind to a number of HLA-DR allotypes. This observation runs counter to the widely-accepted view of MHC class II function, which dictates that each allotype binds a different set of peptides. Furthermore, many if not all of the self-peptides disclosed herein bind to the class II molecules with relatively high affinity. These three characteristics—(1) self rather than foreign, (2) degeneracy, and (3) high affinity binding—suggest a novel means for therapeutic intervention in disease conditions characterized by autoreactivity, such as Type I diabetes, rheumatoid arthritis, and multiple sclerosis. In addition, such therapy could be used to reduce transplant rejection. 
     In the therapeutic methods of the invention, short peptides modelled on the high-affinity immunomodulating self peptides of the invention (which preferably are nonallelically restricted) are introduced into the APCs of a patient. Tissue typing to determine the particular class II alleles expressed by the patient may be unnecessary, as the peptides of the invention are bound by multiple class II isotypes. It may be useful to employ a “cocktail” of peptides, where complete degeneracy is lacking for individual peptides, i.e., where peptides binds to fewer than all allotypes; the cocktail provides overlapping binding specificity. Once in the APC, a peptide binds to the class II molecules with high affinity, thereby blocking the binding of immunogenic peptides which are responsible for the immune reaction characteristic of the disease condition. Because the blocking peptides of the invention are self peptides with the exact carboxy and amino termini tolerized during ontogeny, they are immunologically inert and will not induce an immune response which may complicate treatment using non-self blocking peptides. 
     The peptides of the invention may be introduced into APCs directly, e.g., by intravenous injection of a solution containing one or more of the peptides. Alternatively, the APCs may be provided with a means of synthesizing large quantities of the blocking peptides intracellularly. Recombinant genes that encode ER and/or endosomal targeting signals fused to blocking peptide sequences are linked to appropriate expression control sequences and introduced into APCs. Once in the cell, these genes direct the expression of the hybrid peptides. Peptides targeted to the ER will bind class II α and β chains as they are translated and assembled into heterodimers. The presence of high affinity binding peptides within the ER will prevent association of the α/β complex with invariant chain, and thus interfere with intracellular trafficking. The class II molecule/blocking peptide complex may subsequently be expressed on the cell surface, but would not elicit an immune response since T cells are tolerized to this complex early in development. The use of peptides tagged with ER retention signals may also prevent the peptide-complexed class II molecules from leaving the ER. Alternatively, the recombinant peptide may be tagged with an endosomal targeting signal which directs it to the endosomal compartment after synthesis, thereby also skewing the ratio of endogenously-processed peptide to blocking peptide in the endosome and favoring binding of the high affinity blocking peptide to any class II molecules which did not bind it in the ER. It may be advantageous, for any individual patient, to employ one or more ER-directed peptides in combination with one or more endosome-directed peptide, so that α-β complexes which are not filled in the ER with peptides of the invention are then blocked in the endocytic pathway. The end result again is cell surface expression of a non-immunogenic class II/peptide complex. 
     The use of a class II nonrestricted high affinity binding peptide coupled to an intracellular delivery system permits the specific down-regulation of class II restricted immune responses without invoking the pleiotropic adverse reactions associated with the current pharmacological strategies. Successful application of these technologies will constitute a significant advance towards the treatment of autoimmune disease and prevention of transplant rejection. 
     The intracellular delivery system of the invention can also be utilized in a novel method of vaccination of an animal, e.g., a human patient or a commercially significant mammal such as a cow which is susceptible to diseases such as hoof and mouth disease. Such a system can be tailored to generate the type of immune response required in a given situation by adjustments in the following: (a) peptide specificity for class I or class II MHC; (b) peptide/protein length and/or sequence, and (c) using specific tags for organelle targeting. The system of the invention ensures that peptides are produced only within cells, and are not present outside the cells where they could stimulate antibody production by contact with B cells. This limits the immune response generated by such a vaccine to T cell-mediated immunity, thereby preventing either an inappropriate or potentially deleterious response as might be observed with standard vaccines targeting the organisms which cause, for example, HIV, malaria, leprosy, and leishmaniasis. Furthermore, this exclusively T cell-mediated immune response can be class I or class II-based, or both, depending upon the length and character of the immunogenic peptides: MHC class I molecules are known to bind preferentially to peptides 8 to 10 residues in length, while class II molecules bind with high affinity to peptides that range from 12 to 25 residues long. 
     Immunization and therapy according to the invention can employ a purified preparation of a peptide of the invention, i.e., a peptide which includes an amino acid sequence identical to that of a segment of a naturally-occurring human protein (i.e., a “self protein”), such segment being of 10 to 30 residues in length, wherein the peptide binds to a human MHC class II allotype, and preferably binds to at least two distinct MHC class II allotypes (e.g., any of the approximately 70 known DR allotypes, approximately 47 known DP allotypes, or approximately 33 known DQ allotypes). The portion of the peptide corresponding to the self protein segment is herein termed a “self peptide”. By “purified preparation” is meant a preparation at least 50% (by weight) of the polypeptide constituents of which consists of the peptide of the invention. In preferred embodiments, the peptide of the invention constitutes at least 60% (more preferably at least 80%) of the purified preparation. The naturally-occurring human protein is preferably HLA-A2 (as broadly defined below), HLA-A29, HLA-A30, HLA-B44, HLA-B51, HLA-Bw62, HLA-C, HLA-DP β-chain, HLA-DQ α-chain, HLA-DQ β-chain, HLA-DQ3.2 β-chain, HLA-DR α-chain, HLA-DR β-chain, HLA-DR4 β-chain, invariant chain (Ii), Ig kappa chain, Ig kappa chain C region, Ig heavy chain, Na + /K +  ATPase, potassium channel protein, sodium channel protein, calcium release channel protein, complement C9, glucose-transport protein, CD35, CD45, CD75, vinculin, calgranulin B, kinase C ζ-chain, integrin β-4 gp150, hemoglobin, tubulin α-1 chain, myosin β-heavy chain, α-enolase, transferrin, transferrin receptor, fibronectin receptor α-chain, acetylcholine receptor, interleukin-8 receptor, interferon α-receptor, interferon γ-receptor, calcitonin receptor, LAM (lymphocyte activation marker) Blast-1, LAR (leukocyte antigen-related) protein, LIF (leukemia inhibitory factor) receptor, 4F2 cell-surface antigen (a cell-surface antigen involved in normal and neoplastic growth) heavy chain, cystatin SN, VLA-4 (a cell surface heterodimer in the integrin superfamily of adhesion receptors), PAI-1 (plasminogen activator inhibitor-1), IP-30 (interferon-γ induced protein), ICAM-2, carboxypeptidase E, thromboxane-A synthase, NADH-cytochrome-b5 reductase, c-myc transforming protein, K-ras transforming protein, MET kinase-related transforming protein, interferon-induced guanylate-binding protein, mannose-binding protein, apolipoprotein B-100, cathepsin C, cathepsin E, cathepsin S, Factor VIII, von Willebrand factor, metalloproteinase inhibitor 1 precursor, metalloproteinase inhibitor 2, plasminogen activator inhibitor-1, or heat shock cognate 71 kD protein; it may be an MHC class I or II antigen protein or any other human protein which occurs at the cell surface of APCs. The self peptide preferably conforms to the following motif: at a first reference position (I) at or within 12 residues of the amino terminal residue of the segment, a positively charged residue (i.e., Lys, Arg, or His) or a large hydrophobic residue (i.e., Phe, Trp, Leu, Ile, Met, Tyr, or Pro; and at position I+5, a hydrogen bond donor residue (i.e., Tyr, Asn, Gln, Cys, Asp, Glu, Arg, Ser, Trp, or Thr). In addition, the peptide may also be characterized as having, at positions I+9, I+1, and/or I−1, a hydrophobic residue (i.e., Phe, Trp, Leu, Ile, Met, Pro, Ala, Val, or Tyr) (+ denotes positions to the right, or toward the carboxy terminus, and − denotes positions to the left, or toward the amino terminus.) A typical peptide of the invention will include a sequence corresponding to residues 32-41 (i.e., TQFVRFDSDA; SEQ ID NO: 149) or residues 107-116 (i.e., DWRFLRGYHQ; SEQ ID NO: 150) of HLA-2, or residues 108-117 (i.e., RMATPLLMQA; SEQ ID NO: 151) of Ii, or a sequence essentially identical to any one of the sequences set forth in Tables 1-10 below. 
     The therapeutic and immunization methods of the invention can also employ a nucleic acid molecule (RNA or DNA) encoding a peptide of the invention, but encoding less than all of the entire sequence of the self protein. The nucleic acid preferably encodes no substantial portion of the self protein other than the specified self peptide which binds to a MHC class II molecule, although it may optionally include a signal peptide or other trafficking sequence which was derived from the self protein (or from another protein). A trafficking sequence is an amino acid sequence which functions to control intracellular trafficking (directed movement from organelle to organelle or to the cell surface) of a polypeptide to which it is attached. Such trafficking sequences might traffic the polypeptide to ER, a lysosome, or an endosome, and include signal peptides (the amino terminal sequences which direct proteins into the ER during translation), ER retention peptides such as KDEL (SEQ ID NO: 152); and lysosome-targeting peptides such as KFERQ (SEQ ID NO: 153), QREFK (SEQ ID NO: 154), and other pentapeptides having Q flanked on one side by four residues selected from K, R, D, E, F, I, V, and L. An example of a signal peptide that is useful in the invention is a signal peptide substantially identical to that of an MHC subunit such as class II α or β; e.g., the signal peptide of MHC class II α is contained in the sequence MAISGVPVLGFFIIAVLMSAQESWA (SEQ ID NO: 155). The signal peptide encoded by the nucleic acid of the invention may include only a portion (e.g., at least ten amino acid residues) of the specified 25 residue sequence, provided that portion is sufficient to cause trafficking of the polypeptide to the ER. In preferred embodiments, the nucleic acid of the invention encodes a second self peptide and a second trafficking sequence (which may be identical to or different than the first self peptide and first trafficking sequence), and it may encode additional self peptides and trafficking sequences as well. In still another variation on this aspect of the invention, the self peptide sequence (or a plurality of self peptide sequences arranged in tandem) is linked by a peptide bond to a substantially intact Ii polypeptide, which then carries the self peptide sequence along as it traffics the class II molecule from ER to endosome. 
     The nucleic acid of the invention may also contain expression control sequences (defined as transcription and translation start signals, promoters, and enhancers which permit and/or optimize expression of the coding sequence with which they are associated) and/or genomic nucleic acid of a phage or a virus, such as an attenuated or non-replicative, non-virulent form of vaccinia virus, adenovirus, Epstein-Barr virus, or a retrovirus. 
     The peptides and nucleic acids of the invention may be prepared for therapeutic use by suspending them directly in a pharmaceutically acceptable carrier, or by encapsulating them in liposomes, immune-stimulating complexes (ISCOMS), or the like. Such preparations are useful for inhibiting an immune response in a human patient, by contacting a plurality of the patient&#39;s APCs with the therapeutic preparation and thereby introducing the peptide or nucleic acid into the APCS. 
     Also within the invention is a cell (e.g., a tissue culture cell or a cell, such as a B cell or APC, within a human) containing the nucleic acid molecule of the invention. A cultured cell containing the nucleic acid of the invention may be used to manufacture the peptide of the invention, in a method which involves culturing the cell under conditions permitting expression of the peptide from the nucleic acid molecule. 
     Disclosed herein is a method of identifying a nonallelically restricted immunomodulating peptide, which method includes the steps of: 
     (a) fractionating a mixture of peptides eluted from a first MHC class II allotype; 
     (b) identifying a self peptide from this mixture; and 
     (c) testing whether the self peptide binds to a second MHC class II allotype, such binding being an indication that the self peptide is a nonallelically restricted immunomodulating peptide. 
     In further embodiments, the invention includes a method of identifying a potential immunomodulating peptide, in a method including the steps of: 
     (a) providing a cell expressing MHC class II molecules on its surface; 
     (b) introducing into the cell a nucleic acid encoding a candidate peptide; and 
     (c) determining whether the proportion of class II molecules which are bound to the candidate peptide is increased in the presence of the nucleic acid compared to the proportion bound in the absence of the nucleic acid, such an increase being an indication that the candidate peptide is a potential immunomodulating peptide. 
     Also within the invention is a method of identifying a potential immunomodulating peptide, which method includes the steps of: 
     (a) providing a cell expressing MHC class II molecules on its surface; 
     (b) introducing into the cell a nucleic acid encoding a candidate peptide; and 
     (c) determining whether the level of MHC class II molecules on the surface of the cell is decreased in the presence of the nucleic acid compared to the level of MHC class II molecules in the absence of the nucleic acid, such a decrease being an indication that the candidate peptide is a potential immunomodulating peptide. 
     Also included in the invention is a method of identifying a nonallelically restricted immunostimulating peptide, which method includes the steps of: 
     (a) providing a cell bearing a first MHC class I or class II allotype, such cell being infected with a pathogen (e.g., an infective agent which causes human or animal disease, such as human immunodeficiency virus (HIV), hepatitis B virus, measles virus, rubella virus, influenza virus, rabies virus,  Corynebacterium diphtheriae, Bordetella pertussis , Plasmodium spp., Schistosoma spp., Leishmania spp., Trypanasoma spp., or  Mycobacterium lepre ); 
     (b) eluting a mixture of peptides bound to the cell&#39;s first MHC allotype; 
     (c) identifying a candidate peptide from the mixture, such candidate peptide being a fragment of a protein from the pathogen; and 
     (d) testing whether the candidate peptide binds to a second MHC allotype, such binding being an indication that the candidate peptide is a nonallelically restricted immunostimulating peptide. A nucleic acid encoding such an immunogenic fragment of a protein of a pathogen can be used in a method of inducing an immune response in a human patient, which method involves introducing the nucleic acid into an APC of the patient. 
     The therapeutic methods of the invention solve certain problems associated with prior art methods involving intravenous injection of synthetic peptides: (1) because of allelic specificity, a peptide capable of binding with high affinity to all, or even most, of the different class II allotypes expressed within the general population had not previously been identified; (2) the half-lives of peptides delivered intravenously are generally very low, necessitating repeated administration with the associated high level of inconvenience and cost; (3) this type of delivery approach requires that the blocking peptide displace the naturally-occurring peptide occupying the binding cleft of a class II molecule while the latter is on the cell surface, which is now believed to be a very inefficient process; and (4) if the blocking peptide utilized is itself immunogenic, it may promote deleterious immune responses in some patients. 
     Other features and advantages of the invention will be apparent from the following detailed description, and from the claims. 
    
    
     DETAILED DESCRIPTION 
     The drawings are first briefly described. 
     Drawings 
     FIGS. 1A-1F are chromatographic analyses of the peptide pools extracted from papain digested HLA-DR1, DR2, DR3, DR4, DR7, and DR8, respectively, illustrating the peptide repertoire of each HLA-DR as detected by UV absorbance. The UV absorbance for both 210 nm and 277 nm is shown at a full scale absorbance of 500 mAU with a retention window between 16 minutes and 90 minutes (each mark represents 2 minutes). 
     FIG. 2 is a representative mass spectrometric analysis of the size distribution of isolated HLA-DR1 bound peptides. The determined peptide masses in groups of 100 mass units were plotted against the number of isolated peptides identified by mass spectrometry. Peptide length was calculated by dividing the experimental mass by an average amino acid mass of 118 daltons. 
     FIG. 3A is a representation of a minigene of the invention (SEQ ID NO: 147), in which the HLA-DRα chain leader peptide is linked to the amino terminus of a 15-residue blocking peptide fragment of human invariant chain Ii. 
     FIG. 3B is a representation of a second minigene of the invention (SEQ ID NO: 148), in which the HLA-DRα chain leader peptide is linked to the amino terminus of a 24-residue blocking peptide fragment of human invariant chain Ii. 
    
    
     EXPERIMENTAL DATA 
     METHODS 
     I. Purification of HLA-DR Antigens 
     HLA-DR molecules were purified from homozygous, Epstein-Barr virus-transformed, human B lymphoblastoid lines: DR1 from LG-2 cells, DR2 from MST cells, DR3 from WT20 cells, DR4 from Priess cells, DR7 from Mann cells, and DR8 from 23.1 cells. All of these cell lines are publicly available. Cell growth, harvest conditions and protein purification were as previously described (Gorga, J. et al., 1991). Briefly, 200 grams of each cell type was resuspended in 10 mM Tris-HCl, 1 mM dithiothreitol (DTT), 0.1 mM phenylmethylsulfonylflouride (PMSF), pH 8.0, and lysed in a Thomas homogenizer. The nuclei were removed by centrifugation at 4000×g for 5 min and the pellets washed and repelleted until the supernatants were clear. All the supernatants were pooled and the membrane fraction harvested by centrifugation at 175,000×g for 40 min. The pellets were then resuspended in 10 mM Tris-HCl, 1 mM DTT, 1 mM PMSF, 4% NP-40. The unsolubilized membrane material was removed by centrifugation at 175,000×g for 2 hours, and the NP-40 soluble supernatant fraction used in immunoaffinity purification. 
     Detergent soluble HLA-DR was bound to a LB3.1-protein A SEPHAROSE™ agarose gel column (Pharmacia) (Gorga et al., id) and eluted with 100 mM glycine, pH 11.5. Following elution, the sample was immediately neutralized by the addition of Tris-HCl and then dialyzed against 10 mM Tris-HCl, 0.1% deoxycholic acid (DOC). The LB3.1 monoclonal antibody recognizes a conformational determinant present on the nonpolymorphic HLA-DRα chain, and thus recognizes all allotypes of HLA-DR. 
     The transmembrane domain of the DR molecules was removed by papain digestion, and the resulting water-soluble molecule further purified by gel filtration chromatography on an S-200 column equilibrated in 10 mM Tris-HCl, pH 8.0. The purified DR samples were concentrated by ultrafiltration, yield determined by BCA assay, and analyzed by SDS polyacrylamide gel electrophoresis. 
     II. Extraction and Fractionation of Bound Peptides 
     Water-soluble, immunoaffinity-purified class II molecules were further purified by high-performance size exclusion chromatography (SEC), in 25 mM N-morpholino ethane sulfonic acid (MES) pH 6.5 and a flowrate of 1 ml/min., to remove any residual small molecular weight contaminants. Next, CENTRICON™ ultrafiltration microconcentrators (molecular weight cutoff 10,000 daltons) (Amicon Corp.) were sequentially washed using SEC buffer and 10% acetic acid prior to spin-concentration of the protein sample (final volume between 100-200 μl). Peptide pools were extracted from chosen class II alleles by the addition of 1 ml of 10% acetic acid for 15 minutes at 70° C. These conditions are sufficient to free bound peptide from class II molecules, yet mild enough to avoid peptide degradation. The peptide pool was separated from the class II molecule after centrifugation through the CENTRICON™ ultrafiltration microconcentrator, with the flow-through containing the previously bound peptides. 
     The collected acid-extracted peptide pool was concentrated in a SPEED-VAC™ vacuum equipped centrifluge (Savant) prior to HPLC separation. Peptides were separated on a microbore C-18 reversed-phase chromatography (RPC) column (Vydac) utilizing the following non-linear gradient protocol at a constant flowrate of 0.15 ml/min.: 0-63 min. 5%-33% buffer B; 63-95 min. 33%-60% buffer B; 95-105 min 60%-80% buffer B, where buffer A was 0.06% trifluoroacetic acid/water and buffer B was 0.055% trifluoroacetic acid/acetonitrile. Chromatographic analysis was monitored at multiple UV wavelengths (210, 254, 277, and 292 nm) simultaneously, permitting spectrophotometric evaluation prior to mass and sequence analyses. Shown in FIG. 1 are chromatograms for each of the six DR peptide pools analyzed. 
     Collected fractions were subsequently analyzed by mass spectrometry and Edman sequencing. 
     III. Analysis of Peptides 
     The spectrophotometric evaluation of the peptides during RPC provides valuable information regarding amino acid composition (contribution of aromatic amino acids) and is used as a screening method for subsequent characterization. Appropriate fractions collected during the RPC separation were next analyzed using a Finnegan-MAT LASERMAT™ matrix-assisted laser-desorption mass spectrometer (MALD-MS) to determine the individual mass values for the predominant peptides. Between 1%-4% of the collected fraction was mixed with matrix (1 μl α-Cyano-4-hydroxycinnamic acid) to achieve mass determination of extracted peptides. The result of this analysis for HLA-DR1 is shown in FIG.  2 . Next, chosen peptide samples were sequenced by automated Edman degradation microsequencing using an ABI 477A protein sequencer (Applied Biosystems) with carboxy-terminal verification provided by mass spectral analysis using the Finnigan-MAT TSQ 700 triple quadruple mass spectrometer equipped with an electro-spray ion source. This parallel analysis ensures complete identity of peptide composition and sequence. Peptide alignment with protein sequences stored in the SWISS-PROT database was performed using the FASTA computer database search program. Set forth in Tables 1-10 are the results of this sequence analysis for each of the DR molecules studied. 
     RESULTS 
     I. HLA-DR1 
     The HLA-DR1 used in this study was papain solubilized to enable the material to be used both for crystallographic and bound peptide analyses. The peptides bound to DR1 were acid extracted and fractionated using RPC. (FIG.  1 ). The absence of any detectable peptidic material following a second extraction/RPC separation verified quantitative peptide extraction. Amino acid analysis (ABI 420A/130A derivatizer/HPLC) of extracted peptide pools demonstrated a 70-80% yield, assuming total occupancy of purified DR1 with a molar equivalent of bound peptides corresponding to the size distribution determined by mass spectrometry (see FIG.  2 ). The RPC profiles obtained from DR1 extractions of multiple independent preparations were reproducible. Furthermore, profiles from either detergent-soluble or papain-solubilized DR1 were equivalent. To confirm that the peptides were in fact identical in detergent-soluble and papain-digested DR1, mass spectrometry and Edman sequencing analyses were performed and revealed identical masses and sequences for analogous fractions from the two preparations. 
     Matrix-assisted laser desorption mass spectrometry (MALD-MS) was used to identify 111 species of unique mass contained within the eluted peptide pool of DR1 with an average size of 18 and a mode of 15 residues (FIG.  2 ). Over 500 additional mass species present within the molecular weight range of 13-25 residues were detected; however, the signal was not sufficient to assign individual masses with confidence. Multiple species of varying mass were detected in fractions corresponding to single RPC peaks indicating co-elution of peptides. To characterize these peptides further, samples were analyzed in parallel on a triple quadruple mass spectrometer equipped with an electrospray ion source (ESI-MS) and by automated Edman degradation microsequencing (Lane et al., J. Prot. Chem. 10:151-160 (1991)). Combining these two techniques permits crucial verification of both the N- and C-terminal amino acids of peptides contained in single fractions. The sequence and mass data acquired for twenty peptides isolated from DR1 are listed in Table 1. All the identified peptides aligned with complete identity to regions of proteins stored in the SWISS-PROT database. 
     Surprisingly, sixteen of the twenty sequenced DR1-bound peptides were 100% identical to regions of the self proteins HLA-A2 and class II-associated invariant chain (Ii), representing at least 26% of the total extracted peptide mass. These isolated peptides varied in length and were truncated at both the N- and C-termini, suggesting that: 1) antigen processing occurs from both ends after binding to DR1, or 2) class II molecules bind antigen from a pool of randomly generated peptides. The yields from the peptide microsequencing indicated that HLA-A2 (FIG. 1) and Ii each represents at least 13% of the total DR1-bound peptides. 
     An additional surprising finding concerned a peptide which, although bound to HLA-DR and 100% homologous with HLA-A2 peptide, was derived from a cell which does not express HLA-A2 protein. Evidently this peptide is derived from a protein containing a region homologous with a region of HLA-A2 protein. Thus, for purposes of this specification, the term “HLA-A2 protein” is intended to include HLA-A2 protein itself, as well as any naturally occurring protein which contains a ten or greater amino acid long region of &gt;80% homology with an HLA-DR-binding peptide derived from HLA-A2. An “HLA-A2 peptide” similarly refers to peptides from any HLA-A2 protein, as broadly defined herein. 
     The other four peptides identified in the DR1 studies were derived from two self proteins, transferrin receptor and the Na + /K +  ATPase, and one exogenous protein, bovine serum fetuin (a protein present in the serum used to fortify the medium which bathes the cells). Each of these peptides occupied only 0.3-0.6% of the total DR1 population, significantly less than either the HLA-A2 or the Ii peptides. It is known that class II molecules en route to the cell surface intersect the pathway of incoming endocytic vesicles. Both recycling membrane proteins and endocytosed exogenous protein travel this common pathway. Hence, the HLA-A2, transferrin receptor, Na + /K +  ATPase and bovine fetuin derived peptides would all encounter DR1 in a similar manner. Ii associates with nascent class II molecules in the endoplasmic reticulum (ER) (Jones et al., Mol. Immunol. 16:51-60 (1978)), preventing antigen binding until the class II/Ii complex arrives at an endocytic compartment (Roche and Cresswell, Nature 345:615-618 (1990)), where Ii undergoes proteolysis (Thomas et al., J. Immunol. 140:2670-2675 (1988); Roche and Cresswell, Proc. Natl. Acad. Sci. USA 88:3150-3154 (1991)), thus allowing peptide binding to proceed. Presumably, the Ii peptides bound to DR1 were generated at this step. 
     Synthetic peptides corresponding to five of the peptides reported in Table 1 were made and their relative binding affinities to DR1 determined. The influenza A hemagglutinin peptide (HA) 307-319 (SEQ ID NO: 24) has been previously described as a high affinity, HLA-DR1 restricted peptide (Roche and Cresswell, J. Immunol. 144:1849-1856 (1990); Rothbard et al., Cell 52:515-523 (1988)), and was thus chosen as the control peptide. “Empty” DR1 purified from insect cells expressing recombinant DR1 cDNA was used in the binding experiments because of its higher binding capacity and 10-fold faster association kinetics than DR1 isolated from human cells (Stern and Wiley, Cell 68:465-477 (1992)). All the synthetic peptides were found to compete well (Ki&lt;100 nM) against the HA peptide (Table 2). At first approximation, the Ii 107-120 peptide (SEQ ID NO: 156) had the highest affinity of all the competitor peptides measured, equivalent to that determined for the control HA peptide. In addition to the Ki determinations, these peptides were found to confer resistance to SDS-induced α-β chain dissociation of “empty” DR1 when analyzed by SDS-PAGE, indicative of stable peptide binding (Sadegh-Nasseri and Germain, Nature 353:167-170 (1991); Dornmair et al., Cold Spring Harbor Symp. Quant. Biol. 54:409-415 (1989); Springer et al., J. Biol. Chem. 252:6201-6207 (1977)). Neither of the two control peptides, β 2 m 52-64 (SEQ ID NO: 26) nor Ii 97-111 (SEQ ID NO: 25), was able to either confer resistance to SDS-induced chain dissociation of DR1 or compete with HA 307-319 (SEQ ID NO: 24) for binding to DR1; both of these peptides lack the putative binding motif reported in this study (see below). 
     A putative DR1 binding motif based on the sequence alignments of the core epitopes (the minimum length) of certain naturally processed peptides is shown in Table 3. The peptides listed in this table include those determined herein for HLA-DR1, as well as a number of peptides identified by others and known to bind DR1 (reference #6 in this table being O&#39;Sullivan et al., J. Immunol. 145:1799-1808, 1990; reference #17, Roche &amp; Cresswell, J. Immunol. 144:1849-1856, 1990; reference #25, Guttinger et al., Intern. Immunol. 3:899-906, 1991; reference #27, Guttinger et al. EMBO J. 7:2555-2558, 1988; and reference #28, Harris et al., J. Immunol. 148:2169-2174, 1992). The key residues proposed in the motif are as follows: a positively charged group is located at the first position, referred to here as the index position for orientation (I); a hydrogen bond donor is located at I+5; and a hydrophobic residue is at I+9. In addition, a hydrophobic residue is often found at I+1 and/or I−1. Every naturally processed peptide sequenced from DR1 conforms to this motif (with the exception of the HLA-A2 peptide 103-116 (SEQ ID NO: 3) that lacks residue I+9). Because the putative motif is not placed in a defined position with respect to the first amino acid and because of the irregular length of bound peptides, it is impossible to deduce a motif from sequencing of peptide pools, as was done for class I molecules (Falk et al., Nature 351:290-296 (1991)). The Ii 97-111 peptide (SEQ ID NO: 25), a negative control peptide used in binding experiments, has the I and I+5 motif residues within its sequence, but is missing eight additional amino acids found in Ii 106-119 (SEQ ID NO: 16) (Table 3C). 
     A sequence comparison of 35 previously described DR1-binding synthetic peptides (O&#39;Sullivan et al., J. Immunol. 145:1799-1808 (1990); Guttinger et al., Intern. Immunol. 3:899-906 (1991); Hill et al., J. Immunol. 147:189-197 (1991); Guttinger et al., EMBO J. 7:2555-2558 (1988); Harris et al., J. Immunol. 148:2169-2174 (1992)) also supports this motif. Of the 35 synthetic peptides, 21 (60%) have the precise motif, nine (30%) contain a single shift at either I or I+9, and the remaining five (10%) have a single substitution at I (Table 3B and C). Interestingly, in the latter peptides, a positive charge at I is always replaced by a large hydrophobic residue (Table 8C); a pocket has been described in class I molecules that can accommodate this precise substitution (Latron et al., Proc. Natl. Acad. Sci. USA 88:11325-11329 (1991)). Contributions by the other eight amino acids within the motif or the length of the peptide have not been fully evaluated and may compensate for shifted/missing residues in those peptides exhibiting binding. Evaluation of the remaining 117 non-DR1 binding peptides cited in those studies (which peptides are not included in Table 3) indicates that 99 (85%) of these peptides do not contain the DR1 motif proposed herein. Of the remaining 18 peptides (15%) that do not bind to DR1 but which do contain the motif, 6 (5%) are known to bind to other DR allotypes; the remaining 12 peptides may have unfavorable interactions at other positions which interfere with binding. 
     In contrast to the precise N-terminal cleavages observed in the previous study of six peptides bound to the mouse class II antigen termed I-A b  and five bound to mouse I-E b  (Rudensky et al., Nature 3563:622-627 (1991)), the peptides bound to DR1 are heterogeneous at both the N- and C-termini. In contrast to peptides bound to class I molecules, which are predominantly nonamers (Van Bleek and Nathenson, Nature 348:213-216 (1990); Rotzschke et al., Nature 348:252-254 (1990); Jardetzky et al., Nature 353:326-329 (1991); Hunt et al., Science 255:1261-1263 (1992)), class II peptides are larger and display a high degree of heterogeneity both in length and the site of terminal truncation, implying that the mechanisms of processing for class I and class II peptides are substantially different. Furthermore, the present results suggest that class II processing is a stochastic event and that a DR allotype may bind peptides of different lengths from a complex random mixture. The heterogeneity observed may be solely due to protection of bound peptides from further degradation. Thus, class II molecules would play an active role in antigen processing (as previously proposed (Donermeyer and Allen, J. Immunol. 142:1063-1068 (1989)) by protecting the bound peptides from complete degradation. Alternatively, the predominance of 15mers bound to DR1 (as detected by both the MALD-MS and the yields of sequenced peptides) could be the result of trimming of bound peptides. In any event, the absence of detectable amounts of peptides shorter than 13 and longer than 25 residues suggests that there are length constraints intrinsic either to the mechanism of peptide binding or to antigen processing. The predominance of peptides bound to DR1 that are derived from endogenously synthesized proteins, and particularly MHC-related proteins, may result from the evolution of a mechanism for presentation of self peptides in connection with the generation of self tolerance. 
     II. Other HLA-DR Molecules 
     The sequences of naturally processed peptides eluted from each of DR2, DR3, DR4, DR7 and DR8 are shown in Tables 4-8, respectively. In addition to those peptides shown in Table 4, it has been found that DR2 binds to long fragments of HLA-DR2a β-chain and HLA-DR2b β-chain, corresponding to residues 1-126 or 127 of each of those proteins. Presumably, only a short segment of those long fragments is actually bound within the groove of DR2, with the remainder of each fragment protruding from one or both ends of the groove. Table 9 gives sequences of DR1 from another cell line which does not have wild-type Ar, but which has bound A2-like peptides. Table 10 gives sequences of peptides eluted from DR4 and DR11 molecules expressed in cells from a human spleen. These data demonstrate the great prevalence of self peptides bound, compared to exogenous peptides. The data also show that the A2 and Ii peptides occur repeatedly. In addition, certain of the Tables include peptides that appear to derive from viral proteins, such as Epstein-Barr virus major capsid protein, which are likely to be present in the cells studied. 
     III. Peptide Delivery 
     Genetic Constructions. 
     In order to prepare genetic constructs for in vivo administration of genes encoding immunomodulatory peptides of the invention, the following procedure is carried out. 
     Overlapping synthetic oligonucleotides were used to generate the leader peptide/blocking peptide mini-genes illustrated in FIG. 3 by PCR amplification from human HLA-DRα and invariant chain cDNA templates. These mini-genes encode the Ii peptide fragments KMRMATPLLMQALPM (or Ii 15 ; SEQ ID NO: 15) and LPKPPKPVSKMRMATPLLMQALPM (or Ii 24 ; SEQ ID NO: 7). The resulting constructs were cloned into pGEM-2 (Promega Corp.) to form the plasmids pGEM-2-α-Ii 15  and pGEM-2-α-Ii 24 , with an upstream T7 promoter for use in the in vitro transcription/translation system described below. 
     For in vivo expression, each mini-gene was subsequently subcloned from the pGEM-2 derivatives into a transfection vector, pHβactin-1-neo (Gunning et al., (1987) Proc. Natl. Acad. Sci. U.S.A. 84:4831), to form the plasmids pHβactin-α-Ii 15  and pHβactin-α-Ii 24 . The inserted mini-genes are thus expressed in vivo from the constitutive/strong human β actin promoter. In addition, the mini-genes were subcloned from the pGEM-2 derivatives into the vaccinia virus recombination vector pSC11 (S. Chakrabarti et al. (1985) Mol. Cell. Biol. 5, 3403-3409) to form the plasmids pSC11-α-Ii 15  and pSC11-α-Ii 24 . Following recombination into the viral genome the inserted mini-genes are expressed from the strong vaccinia P 7.5  promoter. 
     Intracellular Trafficking Signals Added to Peptides. 
     Short amino acid sequences can act as signals to target proteins to specific intracellular compartments. For example, hydrophobic signal peptides are found at the amino terminus of proteins destined for the ER, while the sequence KFERQ (SEQ ID NO: 153) (and other closely related sequences) is known to target intracellular polypeptides to lysosomes, while other sequences target polypeptides to endosomes. In addition, the peptide sequence KDEL (SEQ ID NO: 152) has been shown to act as a retention signal for the ER. Each of these signal peptides, or a combination thereof, can be used to traffic the immunomodulating peptides of the invention as desired. For example, a construct encoding a given immunomodulating peptide linked to an ER-targeting signal peptide would direct the peptide to the ER, where it would bind to the class II molecule as it is assembled, preventing the binding of intact Ii which is essential for trafficking. Alternatively, a construct can be made in which an ER retention signal on the peptide would help prevent the class II molecule from ever leaving the ER. If instead a peptide of the invention is targeted to the endosomic compartment, this would ensure that large quantities of the peptide are present when invariant chain is replaced by processed peptides, thereby increasing the likelihood that the peptide incorporated into the class II complex is the high-affinity peptides of the invention rather than naturally-occurring, potentially immunogenic peptides. The likelihood of peptides of the invention being available incorporation into class II can be increased by linking the peptides to an intact Ii polypeptide sequence. Since Ii is known to traffic class II molecules to the endosomes, the hybrid Ii would carry one or more copies of the peptide of the invention along with the class II molecule; once in the endosome, the hybrid Ii would be degraded by normal endosomal processes to yield both multiple copies of the peptide of the invention or molecules similar to it, and an open class II binding cleft. DNAs encoding immunomodulatory peptides containing targeting signals will be generated by PCR or other standard genetic engineering or synthetic techniques, and the ability of these peptides to associate with DR molecules will be analyzed in vitro and in vivo, as described below. 
     It is proposed that the invariant chain prevents class II molecules from binding peptides in the ER and may contribute to heterodimer formation. Any mechanism that prevents this association would increase the effectiveness of class II blockade. Therefore, a peptide corresponding to the site on Ii which binds to the class II heterodimer, or corresponding to the site on either the α or β subunit of the heterodimer which binds to Ii, could be used to prevent this association and thereby disrupt MHC class II function. 
     In Vitro Assembly. 
     Cell free extracts are used routinely for expressing eukaryotic proteins (Krieg, P. &amp; Melton, D. (1984) Nucl. Acids Res. 12, 7057; Pelham, H. and Jackson, R. (1976) Eur. J. Biochem. 67, 247). Specific mRNAs are transcribed from DNA vectors containing viral RNA polymerase promoters (Melton, D. et al. (1984) Nucl. Acids Res. 12, 7035), and added to micrococcal nuclease-treated cell extracts. The addition of  35 S methionine and amino acids initiates translation of the exogenous mRNA, resulting in labeled protein. Proteins may be subsequently analyzed by SDS-PAGE and detected by autoradiography. Processing events such as signal peptide cleavage and core glycosylation are initiated by the addition of microsomal vesicles during translation (Walter, P. and Blobel, G. (1983), Meth. Enzymol., 96, 50), and these events are monitored by the altered mobility of the proteins in SDS-PAGE gels. 
     The ability of peptides containing a signal peptide sequence to be accurately processed and to compete with invariant chain for class II binding in the ER are assayed in the in vitro system described above. Specifically, DR1 α- and β-chain and invariant chain peptide constructs described above are transcribed into mRNAs, which will be translated in the presence of mammalian microsomal membranes. Association of the DR heterodimer with Ii is determined by immunoprecipitation with antisera to DR and Ii. Addition of mRNA encoding the peptide of the invention to the translation reaction should result in a decreased level of coimmunoprecipitated Ii, and the concomitant appearance of coimmunoprecipitated peptide, as determined by SDS-PAGE on TRIS-Tricine gels. These experiments will provide a rapid assay system for determining the potential usefulness of a given blocking peptide as a competitor for Ii chain binding in the ER. Those peptides of the invention which prove to be capable of competing successfully with Ii in this cell-free assay can then be tested in intact cells, as described below. 
     In Vivo Assembly. 
     Human EBV-transformed B cell lines LG-2 and HOM-2 (homozygous for HLA-DR1) and the mouse B cell hybridoma LK35.2 are transfected with either 50 μg of linearized pHβactin-α-Ii 15  or pHβactin-α-Ii 24  or (as a control) pHβactin-1-neo by electroporation (150 mV, 960 μF, 0.2 cm cuvette gap). Following electroporation, the cells are cultured in G418-free medium until total recovery (approximately 4 days). Each population is then placed under G418 selection until neomycin-expressing resistant populations of transfectants are obtained (approximately 1-2 months). The resistant populations are subcloned by limiting dilution and the clonality of stable transfectants determined by PCR amplification of blocking peptide mRNA expression. 
     Stable transfectants of LG-2 and HOM-2 carrying blocking peptide mini-genes or negative control vectors are grown in large-scale culture conditions until 20 grams of pelleted cell mass is obtained. The HLA-DR expressed by each transfectant is purified, and the bound peptide repertoire (both from within the cell and from the cell surface) analyzed as described above. Successful demonstration of a reduction in the total bound peptide diversity will be conclusive evidence of intracellular delivery of immuno-modulatory peptides. 
     A second cell-based assay utilizes stable transfectants of LK35.2 cells carrying blocking peptide mini-genes or negative control vectors; these cells are used as APCs in T cell proliferation assays. Each transfectant is cultured for 24 hours in the presence of different dilutions of hen egg lysozyme (HEL) and HEL-specific T cell hybridomas. The relative activation of the T cells present in each assay (as measured by lymphokine production) is determined using the publicly available lymphokine dependent cell line CTLL2 in a  3 H-thymidine incorporation assay (Vignali et al. (1992) J.E.M. 175:925-932). Successful demonstration of a reduction in the ability of blocking peptide expressing transfectants to present HEL to specific T cell hybridomas will be conclusive evidence of intracellular delivery of immuno-modulatory peptides. Cells of the human TK −  cell line 143 (ATCC) are infected with vaccinia virus (strain WR, TK + ) (ATCC), and two hours postinfection, pSC11-α-Ii 15  or pSC11-α-Ii 24  or pSC11 is introduced into the infected cells by calcium phosphate precipitation. TK −  recombinants are selected for with bromodeoxyuridine at 25 μg/ml. Recombinant plaques are screened by PCR for the presence of mini-gene DNA. Recombinant virus is cloned by three rounds of limiting dilution to generate pure clonal viral stocks. 
     In experiments analogous to the transfection experiments described above, recombinant vaccinia viruses encoding mini-genes or vector alone will be used to infect large-scale cultures of the human EBV transformed B cell lines LG-2 and HOM-2. Following infection, the HLA-DR is purified and the bound peptide repertoire analyzed as described above. A reduction of the complexity of the bound peptide population and a significant increase in the relative amount of Ii peptides bound are conclusive evidence that vaccinia can deliver blocking peptides to human APCs. 
     The same recombinant vaccinia viruses encoding mini-genes or vector will be used to infect mice experiencing experimentally-induced autoimmunity. A number of such models are known and are referred in Kronenberg, Cell 65:537-542 (1991). 
     Liposomal Delivery of Synthetic Peptides or Mini-gene Constructs. 
     Liposomes have been successfully used as drug carriers and more recently in safe and potent adjuvant strategies for malaria vaccination in humans (Fries et al. (1992), Proc. Natl. Acad. Sci. USA 89:358). Encapsulated liposomes have been shown to incorporate soluble proteins and deliver these antigens to cells for both in vitro and in vivo CD8 +  mediated CTL response (Reddy et al., J. Immunol. 148:1585-1589, 1992; and Collins et al., J. Immunol. 148:3336-3341, 1992). Thus, liposomes may be used as a vehicle for delivering synthetic peptides into APCs. 
     Harding et al. (Cell (1991) 64, 393-401) have demonstrated that the targeting of liposome-delivered antigen to either of two intracellular class II-loading compartments, early endosomes and/or lysosomes, can be accomplished by varying the membrane composition of the liposome: acid-sensitive liposomes were found to target their contents to early endosomes, while acid-resistant liposomes were found to deliver their contents to lysosomes. Thus, the peptides of the invention will be incorporated into acid-sensitive liposomes where delivery to endosomes is desired, and into acid-resistant liposomes for delivery to lysosomes. 
     Liposomes are prepared by standard detergent dialysis or dehydration-rehydration methods. For acid-sensitive liposomes, dioleoylphosphatidylethanolamine (DOPE) and palmitoylhomocystein (PHC) are utilized, while dioleoylphospatidylcholine (DOPC) and dioleoylphosphatidylserine (DOPS) are used for the preparation of acid-resistant liposomes. 10 −5  mol of total lipid (DOPC/DOPS or DOPE/PHC at 4:1 mol ratios) are dried, hydrated in 0.2 ml of HEPES buffered saline (HBS) (150 mM NaCl, 1 mM EGTA, 10 mM HEPES pH 7.4) and sonicated. The lipid suspensions are solubilized by the addition of 0.1 ml of 1 M octylglucoside in HBS. The peptides to be entrapped are added to 0.2 ml of 0.6 mM peptide in 20% HBS. The mixture is then frozen, lyophilized overnight, and rehydrated. These liposomes will be treated with chymotrypsin to digest any surface-bound peptide. Liposome delivery to EBV-transformed cell lines (as described above) will be accomplished by 12-16 hour incubation at 37° C. HLA-DR will be purified from the liposome treated cells and bound peptide analyzed as above. 
     Alternatively, the liposomes are formulated with the DNA mini-gene constructs of the invention, and used to deliver the constructs into APCs either in vitro or in vivo. 
     Human immunization will be carried out under the protocol approved by both The Johns Hopkins University Joint Committee for Clinical Investigation and the Human Subject Research Review Board of the Office of the Surgeon General of the U.S. Army (Fries et al. (1992), Proc. Natl. Acad. Sci. U.S.A. 89:358-362), using dosages described therein, or other dosages described in the literature for liposome-based delivery of therapeutic agents. 
     Delivery Via Immune-stimulating Complexes (ISCOMS). 
     ISCOMS are negatively charged cage-like structures of 30-40nm in size formed spontaneously on mixing cholesterol and Quil A (saponin). Protective immunity has been generated in a variety of experimental models of infection, including toxoplasmosis and Epstein-Barr virus-induced tumors, using ISCOMS as the delivery vehicle for antigens (Mowat and Donachie) Immunology Today 12:383-385, 1991. Doses of antigen as low as 1 μg encapsulated in ISCOMS have been found to produce class I mediated CTL responses, where either purified intact HIV-1-IIIB gp 160 envelope glycoprotein or influenza hemagglutinin is the antigen (Takahashi et al., Nature 344:873-875, 1990). Peptides are delivered into tissue culture cells using ISCOMS in a manner and dosage similar to that described above for liposomes; the class II peptide binding of delivered peptides are then determined by extraction and characterization as described above. ISCOM-delivered peptides of the invention which are effectively utilized by cultured cells are then tested in animals or humans. 
     In addition to delivery of the therapeutic synthetic peptides, ISCOMS could be constituted to deliver the mini-gene constructs to APCs, and thus serve as an alternative to the above-outlined vaccinia strategy. 
     Immunogenic Peptide Delivery (Vaccines). 
     In addition to using the above-described intracellular delivery systems to deliver nonimmunogenic self peptides with the specific aim of down-modulating the immune system (thus alleviating autoimmune conditions), the delivery systems of the invention may alternatively be used as a novel means of vaccination, in order to stimulate a portion of the immune system of an animal. In the latter context, the delivery system is employed to deliver, into appropriate cells, DNA constructs which express immunogenic, pathogen-derived peptides intended to stimulate an immune response against a specific pathogen. Because the antigenic peptide is produced inside the target cell itself, the vaccine method of the invention ensures that there is no circulating free antigen available to stimulate antibody formation and thereby induce potentially deleterious or inappropriate immunological reactions. The immune response stimulated by vaccines of the invention is, because the vaccines are targeted solely to APC&#39;s, limited to the T cell mediated response, in contrast to standard vaccine protocols which result in a more generalized immune response. Although some of the peptide-presenting APC&#39;s will initially be lysed by host T cells, such lysis will be limited because, inter alia, the virus-based vaccine is non-replicative, i.e., each carrier virus can infect only one cell. 
     The model antigen that will be used to perfect and test the system of the invention is hen egg lysozyme (HEL). It is arguably the most well characterized protein for antigen presentation studies, to which there are numerous monoclonal antibodies and class I- and class II-restricted mouse T cell clones and hybridomas. The primary epitopes that will be studied are the peptide HEL 34-45, as both monoclonal antibodies and CD4+ T cell hybridomas are available, and peptide HEL 46-61, as both class I and class II-restricted T cell clones and hybridomas have been raised and are publicly available. These two sequences are thus proven immunogenic epitopes. Initially, four constructs encoding different polypeptides are analyzed: (a) whole, secreted HEL, (B) HEL 34-45, (c) HEL 46-61, and (d) HEL 34-61. The last three include a signal sequence known to be cleaved in these cells, e.g., IA k  (MPRSRALILGVLALTTMLSLCGG; SEQ ID NO:274), which would result in targeting to the ER. All constructs are then subcloned into pHβApr-1 neo. The methodology for making these constructs is similar to that outlined above. The constructs are introduced into appropriate APCs, e.g., LK35.2 cells, by means of a conventional eukaryotic transfection or one of the delivery vehicles discussed above (e.g., vaccinia, liposomes, or ISCOMS). LK35.2 cells, which possess the mouse MHC Class II restriction molecules IA k  and IE k , transfected with each of the constructs are tested for their ability to stimulate the appropriate class I and class II-restricted T cell hybridomas and clones using standard techniques. Whether class I stimulation is observed will depend on whether peptide trimming can occur in the ER, in order to produce an 8-10-mer suitable for binding to class I molecules. If these constructs are ineffective for class I stimulation, they can be modified in order to produce a more effective peptide for class I binding. If these constructs prove to be less effective for class II-restricted responses, they can be tagged with endosomal and/or lysosomal targeting sequences as discussed in Section V. 
     The effectiveness of targeting signals used to direct immunogenic peptides to particular intracellular organelles would be monitored using electron microscopic analysis of immunogold stained sections of the various transfectants. Rabbit anti-peptide antisera would be produced and affinity purified for this application. In addition, monoclonal antibody HF10, which recognizes HEL 34-45, will be used. 
     Once a construct is defined that can be effectively presented by transfectants in vitro, its effectiveness in vivo will be determined. This can be tested by injection of the transfectants i.p. and/or s.c. into C3H/Balb/c Fl mice, or by injection of the construct incorporated into an appropriate delivery vehicle (e.g., liposome, ISCOMS, retrovirus, vaccinia). Optimal protocols and doses for such immunizing injections can be determined by one of ordinary skill in the art, given the disclosures provided herein. Efficiency of immunization can be tested by standard methods such as (a) proliferation of class II-restricted T cells in response to HEL pulsed APCs, (b) CTL response to  51 Cr-labeled targets, and (c) serum antibody titre as determined by ELISA. 
     Once the details of the vaccine delivery system of the invention are optimized, constructs encoding peptides with useful immunizing potential can be incorporated into the system. Such peptides can be identified by standard means now used to identify immunogenic epitopes on pathogen-derived proteins. For example, candidate peptides for immunization may be determined from antibody and T cell analysis of animals infected with a particular pathogen. In order to obtain a protective and effective anamnestic response, the peptides used for vaccination should ideally be those which are presented with the highest frequency and efficiency upon infection. This could best be determined by using the procedures outlined in the experimental section above to extract and characterize the peptides bound by MHC class II molecules from infected cells. Given allelic restriction of immunogenic peptides (in contrast to the observed degenerate binding of self peptides of invention), a mini-gene encoding several immunogenic peptides will probably be required to provide a vaccine useful for the entire population. Vaccine administration and dosage are as currently employed to smallpox vaccination. 
     
       
         
               
             
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 LG-2/HLA-DR1 BINDING PEPTIDES 
               
             
          
           
               
                 PROTEIN 
                   
                   
                 SEQ 
                   
                   
                   
                   
                   
               
               
                 SOURCE 
                 POSITION 
                 SEQUENCE 
                 ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
                 YIELD 
               
               
                   
               
             
          
           
               
                 Pseudo HLA-A2 
                 103-120 
                 VGSDWRFLRGYHQYAYDG 
                 1 
                 18 
                 DR1S-59 
                 2190.4 
                 2190.4 
                 39.5 
               
               
                   
                 103-117 
                 VGSDWRFLRGYHQYA 
                 2 
                 15 
                 DR1S-58 
                 1855.0 
                 1854.4 
                 907.5 
               
               
                   
                 103-116 
                 VGSDWRFLRGYHQY 
                 3 
                 14 
                 DR1S-58 
                 1784.0 
                 1783.6 
                 53.3 
               
               
                   
                 104-117 
                 GSDWRFLRGYHQYA 
                 4 
                 14 
                 DR1S-56 
                 1755.3 
                 1755.2 
                 96.5 
               
               
                   
                 105-117 
                 SDWRFLRGYHQYA 
                 5 
                 13 
                 DR1S-56 
                 1698.2 
                 1698.8 
                 48.8 
               
               
                 Invariant Chain 
                  98-122 
                 LPKPPKPVSKMRMATPLLMQALPMG 
                 6 
                 25 
                 DR1S-88 
                 2733.5 
                 2734.5 
                 40.5 
               
               
                 (Ii) 
                  98-121 
                 LPKPPKPVSKMRMATPLLMQALPM 
                 7 
                 24 
                 DR1S-88 
                 2676.4 
                 2675.9 
                 80.8 
               
               
                   
                  99-122 
                 PKPPKPVSKMRMATPLLMQALPMG 
                 8 
                 24 
                 DR1S-86 
                 2620.Z 
                 2619.7 
                 91.5 
               
               
                   
                  98-120 
                 LPKPPKPVSKMRMATPLLMQALP 
                 9 
                 23 
                 DR1S-86 
                 2545.2 
                 2544.5 
                 112.2 
               
               
                   
                  99-121 
                 PKPPKPVSKMRMATPLLMQALPM 
                 10 
                 23 
                 DR1S-87 
                 2563.2 
                 2562.3 
                 145.0 
               
               
                   
                 100-121 
                 KPPKPVSKMRMATPLLMQALPM 
                 11 
                 22 
                 DR1S-87 
                 2466.1 
                 2465.8 
                 101.5 
               
               
                   
                  99-120 
                 PKPPKPVSKMRMATPLLMQALP 
                 12 
                 22 
                 DR1S-84 
                 2432.0 
                 2431.7 
                 72.5 
               
               
                   
                 100-120 
                 KPPKPVSKMRMATPLLMQALP 
                 13 
                 21 
                 DR1S-84 
                 2334.9 
                 2334.2 
                 31.6 
               
               
                   
                 100-120 
                 PPKPVSKMRMATPLLNQALP 
                 14 
                 20 
                 DR1S-86 
                 2206.7 
                 2207.4 
                 89.8 
               
               
                   
                 107-121 
                 KMRMATPLLMQALPM 
                 15 
                 15 
                 DR1S-88 
                 1732.2 
                 1731.9 
                 178.5 
               
               
                   
                 107-120 
                 KMRMATPLLMQALP 
                 16 
                 14 
                 DR1S-86 
                 1601.0 
                 1600.2 
                 162.0 
               
               
                 Na+/K+ ATPase 
                 199-216 
                 IPADLRIISANGCKVDNS 
                 17 
                 18 
                 DR1S-56 
                 1886.6 
                 1885.8 
                 48.8 
               
               
                 Transferrin Recpt. 
                 680-696 
                 RVEYHFLSPYVSPKESP 
                 18 
                 17 
                 DR1S-58 
                 2035.3 
                 2036.8 
                 30.3 
               
               
                 Bovine Fetuin 
                 56-74 
                 YKHTLNQIDSVKVWPRRPT 
                 19 
                 19 
                 DR1S-51 
                 2237.6 
                 2236.5 
                 69.0 
               
               
                   
                 56-73 
                 YKHTLNQIDSVKVWPRRP 
                 20 
                 18 
                 DR1S-50 
                 2338.7 
                 2338.5 
                 32.5 
               
               
                 HLA-DR β-chain 
                 43-61 
                 DVGEYRAVTELGRPDAEYW 
                 21 
                 19 
                 DR1S-51 
                 2226.5 
                 ? 
               
               
                 Carboxypeptidase E 
                 101-115 
                 EPGEPEFKYIGNMHG 
                 22 
                 15 
                 DR1S-48 
                 1704.9 
                 1700.4* 
               
               
                   
                   
                   
                   
                   
                   
                   
                 ESI-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 PEPTIDE BINDING TO MLA-DR1 
               
             
          
           
               
                   
                   
                   
                 Ki vs HA 307-319 b   
                 SDS-Resistance c   
               
               
                 PEPTIDE a   
                 SEQ ID NO. 
                 LENGTH 
                 nM 
                 nM 
               
               
                   
               
             
          
           
               
                 HLA-A2 103-117 
                 2 
                 15 
                 49 ± 3  
                 + 
               
               
                 Ii 106-120 
                 15 
                 15 
                 &lt;10 
                 + 
               
               
                 Ii 98-121 
                 7 
                 24 
                 33 ± 5  
                 + 
               
               
                 Na+/K+ ATPase 199-216 
                 17 
                 18 
                 68 ± 9  
                 + 
               
               
                 Transf. Recept. 680-696 
                 18 
                 17 
                 &lt;10 
                 + 
               
               
                 Bovine Fetuin 56-72 
                 23 
                 19 
                 66 ± 18 
                 + 
               
               
                 HA 307-319 
                 24 
                 14 
                 &lt;10 
                 + 
               
               
                 Ii 97-111 
                 25 
                 15 
                     &gt;10 4   
                 − 
               
               
                 β 2 m 52-64 
                 26 
                 13 
                     &gt;10 4   
                 — 
               
               
                   
               
               
                   a  The first six entries correspond to peptides found associated with HLA-DR1 and the sequences are shown in Table 1. Two control peptides were also tested: β 2 m 52-64, SDLSFSKDWSFYL (SEQ ID NO: 26), is from human β 2 -microglobulin and Ii 97-111, LPKPPKPVSKMRMAT (SEQ ID NO: 25) is a truncated version of the longest invariant chain derived peptide isolated from HLA-DR1.  
               
               
                 # Peptides were synthesized using solid-phase Fmoc chemistry, deprotected and cleaved using standrd methods, then purified by RPC. Purfied peptides were analyzed by mass spectrometry and concentrations were determined by quantitative ninhydrin analysis.  
               
               
                   b Inhibition constants (Ki) were measured as the concentration of test peptide which inhibited 50% of the  125 I-labeled HA 307-319 binding to “empty” HLA-DR1 produced in Sf9 insect cells (20). HA 307-319 was labeled using Na  125 I and chloramine-T and isolated by gel filtration. Specific activity, determined by BCA assay (Pierce) and gamma counting, was 26,000 cpm/pmol.  
               
               
                 # 10 nM labeled peptide and 10 nM purified HLA-DR1 were mixed with 10 different concentrations (10 nM to 10 μM) of synthetic cold competitor peptide in phosphate-buffered saline, pH 7.2, containing 1 mM EDTA, 1 mM PMSF, 0.1 mM iodoacetamide, and 3 mM NaN 3 , and incubated at 37° C. for 85 hours. Free and bound peptfde were separated by native gel electrophoresis (33) and bound radioactivity was quantitated using a  
               
               
                 # Fujix imaging plate analyzer (BAS 2000) after four hour exposures on the phospho-imaging plates. Percent inhibition was calculated as the ratio of background-corrected radioactivity in the sample to background-corrected radioactivity in a parallel sample containing no competitor peptide. Under these conditions, Ki measurements &lt;10 nM could not be accurately determined.  
               
               
                   c The ability of the synthetic peptides to confer resistance to SDS-induced chain dissociation of HLA-DR1 produced in insect cells was determined as described (20). Briefly, 20 μM HLA-DR1 was incubated with five-fold excess of synthetic peptide at 37° C. for 85 hours, in phosphate-buffered saline (pH 7.2) with the protease inhibitor mixture described above.  
               
               
                 # After incubation, the samples were analyzed by SDS-PAGE with and without boiling prior to loading. Peptides which prevented SDS-induced chain dissociation are indicated positive (+) and those that did not negative (−).  
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 PUTATIVE HLA-DR1 PEPTIDE BINDING MOTIF 
               
             
          
           
               
                 A 
                 PROTEIN SOURCE 
                 PEPTIDE SEQUENCE 
                 SEQ ID NO 
                 LENGTH 
                 POSITION 
                 REFERENCE 
               
               
                   
               
             
          
           
               
                   
                 HLA-A2 
                 SDW R FLRG Y HQY A   
                 5 
                 13 
                 105-117 
                 This study 
               
               
                   
                 Invariant Chain 
                   K MRMA T PLL M QALP 
                 16 
                 14 
                 105-118 
                   
               
               
                   
                 Na+/K+ ATPase 
                 IPADL R IISA N GCK V DNS 
                 17 
                 18 
                 199-216 
                   
               
               
                   
                 Transferrin Receptor 
                 RVEY H FLSP Y VS P KESP 
                 18 
                 17 
                 680-696 
                   
               
               
                   
                 Bovine Fetuin 
                 Y K HTLN Q IDS V KVWPRRP 
                 20 
                 18 
                 56-73 
                   
               
               
                 B 
                 HEL 
                   K VFGR C ELA A AMKRHGLD 
                 27 
                 18 
                  1-18 
                 6 
               
               
                   
                   
                 RN R CKGT D VQA W IRGCRL 
                 28 
                 18 
                 112-129 
                 6 
               
               
                   
                 β 2 m 
                   H PPHI E IQM L KNGKKI 
                 29 
                 16 
                 31-46 
                 6 
               
               
                   
                 PLA 2   
                 NELGRF K HTDA C CRT H   
                 30 
                 16 
                 19-34 
                 6 
               
               
                   
                   
                 S K PKVY Q WFD L RKY 
                 31 
                 14 
                 115-128 
                 6 
               
               
                   
                 NASE 
                 ATST K KLHK E PAT L IKAIDG 
                 32 
                 20 
                  1-20 
                 6 
               
               
                   
                   
                 PATLI K AIDG D TVK L MYKGQ 
                 33 
                 20 
                 11-30 
                 6 
               
               
                   
                   
                 DRVKLMY K GQPM T FRL L LVD 
                 34 
                 20 
                 21-40 
                 6 
               
               
                   
                   
                 VAYVYKPNNT H EQHL R KSE A   
                 35 
                 20 
                 111-130 
                 6 
               
               
                   
                 HIV p13 
                 Q K QEPI D KEL Y PLTSL 
                 36 
                 16 
                  97-112 
                 6 
               
               
                   
                 HIV p17 
                 GA R ASVL S GGE L DKWE 
                 37 
                 16 
                  1-16 
                 6 
               
               
                   
                 Influenza HA 
                   R TLYQ N VGT Y VSVGTSTLNK 
                 38 
                 20 
                 187-206 
                 6 
               
               
                   
                 Influenza HA 
                 P K YVKQ N TLK L AT 
                 24 
                 13 
                 307-319 
                 17 
               
               
                   
                 P. falcip. p190 
                 LK K LVFG Y RKP L DNI 
                 39 
                 15 
                 249-263 
                 25 
               
               
                   
                 P. falcip. CS 
                   K HIEQ Y LKK I KNS 
                 40 
                 13 
                 329-341 
                 27 
               
               
                   
                 Chicken OVA 
                 DVFKEL K VHHA N ENI F   
                 41 
                 16 
                 15-30 
                 6 
               
               
                   
                 DR1 β chain 
                 GDT R PRFL W QLK F ECHFFNG 
                 42 
                 20 
                  1-20 
                 28 
               
               
                   
                   
                 TERVRLLE R CIYN Q EES V RFDS 
                 43 
                 22 
                 21-42 
                 28 
               
               
                   
                   
                 DLLEQR R AAVD T YCR H NYGVGESFT 
                 44 
                 25 
                 66-90 
                 28 
               
               
                   
                 p Cyt c 
                   K AERA D LIA Y LKQATAK 
                 45 
                 17 
                  88-104 
                 6 
               
               
                   
                 Myelin basic prot. 
                 G R TQDE N PVV H FFKNIVTPRTPPP 
                 46 
                 24 
                 75-98 
                 6 
               
               
                 C 
                 Influenza Matrix 
                 PL K AEIA Q RLEDV 
                 47 
                 13 
                 19-31 
                 6 
               
               
                   
                 HIV p17 
                   R QILG Q LQP S LQTGSE 
                 48 
                 16 
                 57-72 
                 6 
               
               
                   
                 β 2 M 
                 IQVYS R HPPE N GKP N I 
                 49 
                 16 
                  7-22 
                 6 
               
               
                   
                 PLA 2   
                 INT K CYKL E HPV T GCG 
                 50 
                 16 
                  85-100 
                 6 
               
               
                   
                 P. falcip p190 
                   Y KLNF Y FDL L RAKL 
                 51 
                 14 
                 211-224 
                 25 
               
               
                   
                   
                 IDT L KKNE N IKEL 
                 52 
                 13 
                 338-350 
                 25 
               
               
                   
                 DR1 β chain 
                 DVGE Y RAVT E LGR P DAEYWN 
                 53 
                 20 
                 43-62 
                 28 
               
               
                   
                 HIV p17 
                   E RFAV N PGL L ETSEGC 
                 54 
                 16 
                 41-56 
                 6 
               
               
                   
                 HEL 
                 DNYR G YSLG N WVC A AKFESNFTQ 
                 55 
                 23 
                 20-42 
                 6 
               
               
                   
                 NASE 
                 EALVRQG L AKVA Y VYK P NNT 
                 56 
                 20 
                 101-120 
                 6 
               
               
                   
                 HIV p25 
                 P I VQNL Q GQM V HQAIS 
                 57 
                 16 
                  1-16 
                 6 
               
               
                   
                   
                 SA L SEGA T PQD L NTML 
                 58 
                 16 
                 41-56 
                 6 
               
               
                   
                 β 2 m 
                 SF Y ILAH T EFT P TETD 
                 59 
                 16 
                 61-76 
                 6 
               
               
                   
                 PLA 2   
                 KM Y FNLI N TKC Y KLEH 
                 60 
                 16 
                 79-94 
                 6 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 MST/HLA-DR2 BINDING PEPTIES 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 Pseudo HLA-A2 
                 103-120 
                 VGSDWRFLRGYHQYAYDG 
                 1 
                 18 
                 DR2-3-57 
                 2190.4 
                 2189.0 
               
               
                   
                 103-119 
                 VGSDWRFLRGYHQYAYAD 
                 61 
                 17 
                 DR2-3-57 
                 2133.3 
                 2131.8 
               
               
                   
                 104-119 
                 GSDWRFLRGYHQYAYD 
                 62 
                 16 
                 DR2-3-56 
                 2034.3 
                 2040.4 
               
               
                   
                 103-117 
                 VGSDWRFLRGYHQYA 
                 2 
                 15 
                 DR2-3-56 
                 1855.0 
                 1858.5 
               
               
                   
                 103-116 
                 VGSDWRFLRGYHQY 
                 3 
                 14 
                 DR2-3-56 
                 1784.0 
                 1786.3 
               
               
                   
                 104-117 
                 GSDWRFLRGYHQYA 
                 4 
                 14 
                 DR2-3-55 
                 1755.3 
                 1755.0* 
               
               
                   
                 105-117 
                 SDWRFLRGYHQYA 
                 5 
                 13 
                 DR2-3-56 
                 1698.2 
                 1702.6 
               
               
                 Invariant Chain 
                  98-121 
                 LPKPPKPVSKMRMATPLLMQALPM 
                 7 
                 24 
                 DR2-3-70 
                 2676.4 
                 2675.0* 
               
               
                 (Ii) 
                  99-121 
                 PKPPKPVSKMRMATPLLMQALPM 
                 10 
                 23 
                 DR2-3-70 
                 2563.2 
                 2562.0* 
               
               
                   
                 100-121 
                 KPPKPVSKMRMATPLLMQALPM 
                 11 
                 22 
                 DR2-3-70 
                 2466.1 
                 2465.0* 
               
               
                   
                  99-120 
                 PKPPKPVSKMRMATPLLMQALP 
                 12 
                 22 
                 DR2-3-66 
                 2432.0 
                 2437.0 
               
               
                   
                 100-120 
                 KPPKPVSKMRMATPLLMQALP 
                 13 
                 21 
                 DR2-3-66 
                 2334.9 
                 2340.0 
               
               
                   
                 101-120 
                 PPKPVSKMRMATPLLNQALP 
                 63 
                 20 
                 DR2-3-70 
                 2206.7 
                 2207.0* 
               
               
                   
                 107-125 
                 KMRMATPLLMQALPMGALP 
                 64 
                 19 
                 DR2-3-71 
                 2070.5 
                 2074.3 
               
               
                   
                 107-121 
                 KMRMATPLLMQALPM 
                 15 
                 15 
                 DR2-3-70 
                 1732.2 
                 1732.0* 
               
               
                 HLA-DQ α-chain 
                  97-119 
                 NIVIKRSNSTAATNEVPEVTVFS 
                 158 
                 23 
                 DR2-3-44 
                 2476.8 
                 2478.1 
               
               
                   
                  97-112 
                 NIVIKRSNSTAATNEV 
                 159 
                 16 
                 DR2-3-41 
                 1716.9 
                 1717.0 
               
               
                 HLA-DQ β-chain 
                 42-59 
                 SDVGVYRAVTPQGRPDAE 
                 160 
                 18 
                 DR2-3-41 
                 1917.1 
                 1920.5 
               
               
                   
                 43-59 
                 DVGVYRAVTPQGRPDAE 
                 161 
                 17 
                 DR2-3-41 
                 1830.0 
                 1833.3 
               
               
                   
                 43-57 
                 DVGVYRAVTPQGRPD 
                 162 
                 15 
                 DR2-3-41 
                 1629.8 
                 1632.9 
               
               
                 HLA-DR α-chain 
                 182-194 
                 APSPLPETTENVV 
                 163 
                 13 
                 DR2-3-36 
                 1353.5 
                 1362.0 
               
               
                   
                 182-198 
                 APSPLPETTENVVCALG 
                 164 
                 17 
                 DR2-3-41 
                 1697.9 
                 1701.0 
               
               
                 (MET) Kinase-relate 
                 59-81 
                 EHHIFLGATNYIYVLNEEDLQKV 
                 65 
                 23 
                 DR2-3-65 
                 2746.1 
                 2746.6 
               
               
                 trasforming protein 
               
               
                 Guanylate-bind. 
                 434-450 
                 QELKNKYYQVPRKGIQA 
                 66 
                 17 
                 DR2-3-71 
                 2063.4 
                 2074.3 
               
               
                 Mannose-bind. prot. 
                 174-193 
                 IQNLIKEEAFLGITDEKTEG 
                 67 
                 20 
                 DR2-3-70 
                 2248.5 
                 2248.0* 
               
               
                 Apolipoprotein B-100 
                 1200-1220 
                 FPKSLHTYANILLDRRVPQTD 
                 165 
                 21 
                 DR2-3-61 
                 2484.8 
                 2490.9 
               
               
                   
                 1200-1218 
                 FPKSLHTYANILLDRRVPQ 
                 166 
                 19 
                 DR2-3-61 
                 2268.6 
                 2276.7 
               
               
                 Potassium channel prot 
                 173-190 
                 DGILYYYQSGGRLRRPVN 
                 167 
                 18 
                 DR2-3-61 
                 2127.4 
                 2132.6 
               
               
                   
                 173-189 
                 DGILYYYQSGGRLRRPV 
                 168 
                 17 
                 DR2-3-61 
                 2013.3 
                 2018.1 
               
               
                 Fibronectin receptor 
                 586-616 
                 LSPIHIALNFSLDPQAPVDSHGLRPALHYQ 
                 169 
                 30 
                 DR2-3-61 
                 3307.7 
                 3313.1 
               
               
                 Factor VIII 
                 1175-1790 
                 LWDYGMSSSPHVLRNR 
                 170 
                 16 
                 DR2-3-44 
                 1918.2 
                 1921.7 
               
               
                 HLA-DR2b β-chain 
                  94-111 
                 RVQPKVTVYPSKTQPLQH 
                 72 
                 18 
                 DR2-3-39 
                 2106.5 
                 2114. 
               
               
                   
                  94-108 
                 RVQPKVTVYPSKTQP 
                 73 
                 15 
                 DR2-3-39 
                 1728.3 
                 1730.6 
               
               
                   
                   
                   
                   
                   
                   
                   
                 ESI.MS* 
               
               
                   
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 WT-2-/HLA-DR3 NATURALLY PROCESSED PEPTIDES 
               
             
          
           
               
                 Protein Source 
                 Position 
                 Sequence 
                 SEQ ID NO. 
                 Length 
                 Fraction 
                 MW 
                 Mass Spec. 
               
               
                   
               
             
          
           
               
                 Pseudo HLA-A2 
                 103-117 
                 VGSDWRFLRGYHQYA 
                 2 
                 15 
                 DR3-2-63 
                 1855.0 
                 1863.9 
               
               
                 HLA-A30 
                 28-?  
                 VDDTQFVRFDSDAASQ . . . 
                 171 
                 ? 
                 DR3-2-55 
                 ? 
                 ? 
               
               
                 HLA-DR α-chain 
                 111-129 
                 PPEVTVLTNSPVELREPNV 
                 172 
                 19 
                 DR3-2-55 
                 2090.4 
                 2093.3 
               
               
                   
                 111-128 
                 PPEVTVLTNSPVELREPN 
                 173 
                 18 
                 DR3-2-55 
                 1991.2 
                 1989.8 
               
               
                 HLA-DR β-chain 
                 1-? 
                 GDTRPRFLEYSTSECHFF 
                 79 
                 18 
                 DR3-2-73 
                 ? 
                 ? 
               
               
                 Acetylcholine recept. 
                 289-304 
                 VFLLLLADKVPETSLS 
                 174 
                 16 
                 DR3-2-65 
                 1745.1 
                 1750.1 
               
               
                 Glucose-transport 
                 459-474 
                 TFDEIASGFRQGGASQ 
                 175 
                 16 
                 DR3-2-55 
                 1670.8 
                 1672.6 
               
               
                 Sodium channel prot. 
                 384-397 
                 YGYTSYDTFSWAFL 
                 176 
                 14 
                 DR3-2-41 
                 1720.8 
                 1720.5 
               
               
                 Invariant chain 
                  98-120 
                 LPKPPKPVSKMRMATPLLMQALP 
                 9 
                 23 
                 DR3-2-73 
                 2545.2 
                 2554.0 
               
               
                 (Ii) 
                  99-120 
                 PKPPKPVSKMRMATPLLMQALP 
                 12 
                 22 
                 DR3-2-73 
                 2432.0 
                 2441.4 
               
               
                   
                 100-120 
                 KPPKPVSKMRMATPLLMQALP 
                 13 
                 21 
                 DR3-2-73 
                 2334.9 
                 2345.3 
               
               
                   
                 132-150 
                 ATKYGNMTEDHVMHLLQNA 
                 177 
                 19 
                 DR3-2-69 
                 2173.4 
                 2179.3 
               
               
                 CD45 
                 1071-1084 
                 GQVKKNNHQEDKIE 
                 178 
                 14 
                 DR3-2-41 
                 1666.8 
                 1667.0 
               
               
                 ICAM-2 
                 64-76 
                 LNKILLDEQAQWK 
                 179 
                 13 
                 DR3-2-51/52 
                 1598.9 
                 1602.4 
               
               
                 Interferon γ-receptor 
                 128-147 
                 GPPKLDIRKEEKQIMIDIFH 
                 180 
                 21 
                 DR3-2-77 
                 2505.0 
                 2510.3 
               
               
                   
                 128-148 
                 GPPKLDIRKEEKQIMIDIFHP 
                 181 
                 20 
                 DR3-2-77 
                 2407.8 
                 2412.4 
               
               
                 IP-30 
                 38-59 
                 SPLQALDFFGNGPPVNYKTGNL 
                 182 
                 22 
                 DR3-2-77 
                 2505.0 
                 2510.3 
               
               
                   
                 38-57 
                 SPLQALDFFGNGPPVNYKTG 
                 183 
                 20 
                 DR3-2-77 
                 2122.4 
                 2124.2 
               
               
                 Cytochrome-b5 reduc. 
                 155-172 
                 GKFAIRPDKKSNPIIRTV 
                 184 
                 18 
                 DR3-2-51/52 
                 2040.4 
                 2043.2 
               
               
                 EBV membrane antigen 
                 592-606 
                 TGHGARTSTEPTTDY 
                 185 
                 15 
                 DR3-2-41 
                 1593.6 
                 1592.7 
               
               
                 GP220 
               
               
                 EBV tegument protein 
                 1395-1407 
                 KELKRQYEKKLRQ 
                 186 
                 13 
                 DR3-2-51/52 
                 1747.1 
                 1749.8 
               
               
                 membrane p140 
               
               
                 Apolipoprotein 
                 1276-1295 
                 NFLKSDGRIKYTLNKNSLK 
                 74 
                 20 
                 DR3-2-63 
                 2352.9 
                 2360.0 
               
               
                 B-100 (Human) 
                 1273-1292 
                 IPDNLFLKSDGRIKYTLNKN 
                 191 
                 20 
                 DR3-2-65 
                 2349.7 
                 2354.6 
               
               
                   
                 1273-1291 
                 IPDNLFLKSDGRIKYTLNK 
                 75 
                 19 
                 DR3-2-63 
                 2235.5 
                 2245.1 
               
               
                   
                 1273-1290 
                 IPDNLFLKSDGRIKYTLN 
                 192 
                 18 
                 DR3-2-65 
                 2107.4 
                 2096.6 
               
               
                   
                 1273-1289 
                 IPDNLFLKSDGRIKYTL 
                 193 
                 17 
                 DR3-2-65 
                 1993.3 
                 2000.8 
               
               
                   
                 1276-1291 
                 NLFLKSDGRIKYTLNK 
                 76 
                 16 
                 DR3-2-60 
                 1910.2 
                 1911.4 
               
               
                   
                 1276-1290 
                 NLFLKSDGRIKYTLN 
                 77 
                 15 
                 DR3-2-60 
                 1782.1 
                 1785.9 
               
               
                   
                 1207-1224 
                 YANILLDRRVPQTDMTF 
                 78 
                 17 
                 DR3-2-63 
                 2053.3 
                 2059.1 
               
               
                   
                 1794-1810 
                 VTTLNSDLKYNALDLTN 
                 194 
                 17 
                 DR3-2-69 
                 1895.1 
                 1896.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 PRIESS/HLA-DR4 NATURALLY PROCESSED PEPTIDES 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 Ig Kappa Chain 
                 188-208 
                 KHKVYACEVTHQGLSSPVTKS 
                 80 
                 21 
                 DR4-2-45 
                 2299.6 
                 2304.0 
               
               
                 C regIon (Human 
                 188-207 
                 KHKVYACEVTHQGLSSPVTK 
                 81 
                 20 
                 DR4-2-47 
                 2212.5 
                 2213.0 
               
               
                   
                 189-206 
                 HKVYACEVTHQGLSSPVT 
                 82 
                 18 
                 DR4-2-43 
                 1955.5 
                 1952.1 
               
               
                   
                 188-204 
                 KHKVYACEVTHQGLSSP 
                 83 
                 17 
                 DR4-2-45 
                 1883.1 
                 1882.8 
               
               
                   
                 187-203 
                 EKHKVYACEVTHQGLSS 
                 84 
                 17 
                 DR4-2-45 
                 1915.1 
                 1922.5 
               
               
                   
                 188-203 
                 KHKVYACEVTHQGLSS 
                 85 
                 16 
                 DR4-2-54 
                 1787.0 
                 1787.0 
               
               
                   
                 189-204 
                 HKVYACEVTHQGLSSP 
                 86 
                 16 
                 DR4-2-47 
                 1755.0 
                 1767.8 
               
               
                   
                 187-202 
                 EKHKVYACEVTHQGLS 
                 87 
                 16 
                 DR4-2-43 
                 1828.0 
                 1822.8 
               
               
                   
                 188-202 
                 KHKVYACEVTHQGLS 
                 88 
                 15 
                 DR4-2-51 
                 1699.9 
                 1708.3 
               
               
                   
                 189-203 
                 HKVYACEVTHQGLSS 
                 89 
                 15 
                 DR4-2-45 
                 1657.8 
                 1667.0 
               
               
                   
                 187-200 
                 EKHKVYACEVTHQG 
                 90 
                 14 
                 DR4-2-51 
                 1628.8 
                 1632.6 
               
               
                 HLA-DR α-chain 
                 182-198 
                 APSPLPETTENVVCALG 
                 91 
                 17 
                 DR4-2-43 
                 1697.9 
                 1700 
               
               
                 HLA-A2 
                 28-50 
                 VDDTQFVRFDSDAASQRMEPRAP 
                 195 
                 23 
                 DR4-2-58 
                 2638.6 
                 2641.5 
               
               
                   
                 28-48 
                 VDDTQFVRFDSDAASQRMEPR 
                 92 
                 21 
                 DR4-2-56 
                 2470.6 
                 2472.9 
               
               
                   
                 28-47 
                 VDDTQFVRFDSDAASQRMEP 
                 93 
                 20 
                 DR4-2-59 
                 2314.5 
                 2319.3 
               
               
                   
                 28-46 
                 VDDTQFVRFDSDAASQRME 
                 94 
                 19 
                 DR4-2-54 
                 2217.2 
                 2218.7 
               
               
                   
                 30-48 
                 DTQFVRFDSDAASQRMEPR 
                 95 
                 19 
                 DR4-2-55 
                 2256.4 
                 2263.2 
               
               
                   
                 31-49 
                 TQFVRFDSDAASQRMEPRA 
                 96 
                 19 
                 DR4-2-56 
                 2212.4 
                 2211.5 
               
               
                   
                 28-44 
                 VDDTQFVRFDSDAASQR 
                 97 
                 17 
                 DR4-2-55 
                 1957.0 
                 1963.1 
               
               
                   
                 31-47 
                 TQFVRFDSDAASQRMEP 
                 98 
                 17 
                 DR4-2-56 
                 1985.1 
                 1987.5 
               
               
                   
                 31-45 
                 TQFVRFDSDAASQRM 
                 99 
                 15 
                 DR4-2-54 
                 1758.9 
                 1761.0 
               
               
                   
                 31-42 
                 TQFVRFDSDAAS 
                 100 
                 12 
                 DR4-2-54 
                 1343.4 
                 1343.3 
               
               
                 HLA-C 
                 28-50 
                 VDDTQFVRFDSDAASPRGEPRAP 
                 101 
                 23 
                 DR4-2-56 
                 2533.7 
                 2536.7 
               
               
                   
                 31-52 
                 TQFVRFDSDAASPRGEPRAPWV 
                 102 
                 22 
                 DR4-2-54 
                 2489.7 
                 2491.5 
               
               
                   
                 28-48 
                 VDDTQFVRFDSDAASPRGEPR 
                 103 
                 21 
                 DR4-2-54 
                 2365.5 
                 2368.1 
               
               
                   
                 28-47 
                 VDDTQFVRFDSDAASPRGEP 
                 104 
                 20 
                 DR4-2-56 
                 2209.3 
                 2211.5 
               
               
                   
                 28-46 
                 VDDTQFVRFDSDAASPRGE 
                 105 
                 19 
                 DR4-2-56 
                 2112.2 
                 2113.9 
               
               
                 HLA-Cw9 
                 28-45 
                 VDDTQFVRFDSDAASPRG 
                 106 
                 18 
                 DR4-2-56 
                 1983.1 
                 1987.5 
               
               
                   
                 31-48 
                 TQFVRFDSDAASPRGEPR 
                 107 
                 18 
                 DR4-2-52 
                 2036.2 
                 2041.5 
               
               
                   
                 28-44 
                 VDDTQFVRFDSDAASPR 
                 108 
                 17 
                 DR4-2-55 
                 1926.0 
                 1931.7 
               
               
                   
                 30-46 
                 DTQFVRFDSDAASPRGE 
                 109 
                 17 
                 DR4-2-52 
                 1897.9 
                 1901.6 
               
               
                   
                 31-44 
                 TQFVRFDSDAASPR 
                 110 
                 14 
                 DR4-2-52 
                 1596.7 
                 1603.7 
               
               
                   
                 31-42 
                 TQFVRFDSDAAS 
                 111 
                 12 
                 DR4-2-54 
                 1343.4 
                 1343.3 
               
               
                 HLA-C 
                 130-150 
                 LRSWTAADTAAQITQRKWEAA 
                 112 
                 21 
                 DR4-2-56 
                 2374.6 
                 2376.4 
               
               
                   
                 129-147 
                 DLRSWTAADTAAQITQRKW 
                 197 
                 19 
                 DR4-2-58 
                 2218.4 
                 2220.1 
               
               
                   
                 130-147 
                 LRSWTAADTAAQITQRKW 
                 198 
                 18 
                 DR4-2-58 
                 2103.3 
                 2105.0 
               
               
                   
                 129-145 
                 DLRSWTAADTAAQITQR 
                 113 
                 17 
                 DR4-2-59 
                 1904.5 
                 1908.7 
               
               
                   
                 129-144 
                 DLRSWTAADTAAQITQ 
                 114 
                 16 
                 DR4-2-59 
                 1747.9 
                 1752.3 
               
               
                   
                 129-143 
                 DLRSWTAADTAAQIT 
                 115 
                 15 
                 DR4-2-59 
                 1619.7 
                 1622.2 
               
               
                 HLA-Bw62 
                 129-150 
                 DLSSWTAADTAAQITQRKWEAA 
                 199 
                 22 
                 DR4-2-65 
                 2420.6 
                 2422.7 
               
               
                   
                 129-145 
                 DLSSWTAADTAAQITQR 
                 116 
                 17 
                 DR4-2-60 
                 1834.9 
                 1838.1 
               
               
                   
                 129-146 
                 DLSSWTAADTAAQITQRK 
                 200 
                 18 
                 DR4-2-65 
                 1963.1 
                 1966.3 
               
               
                   
                 129-148 
                 DLSSWTAADTAAQITQRKWE 
                 117 
                 20 
                 DR4-2-66 
                 2278.4 
                 2284.6 
               
               
                 VLA-4 
                 229-248 
                 GSLFVYNITTNKYKAFLDKQ 
                 201 
                 20 
                 DR4-2-65 
                 2350.7 
                 2352.6 
               
               
                   
                 229-244 
                 GSLFVYNITTNKYKAF 
                 202 
                 16 
                 0R4-2-65 
                 1866.1 
                 1868.2 
               
               
                 PAI-1 
                 261-281 
                 AAPYEKEVPLSALTNILSAQL 
                 203 
                 21 
                 DR4-2-65 
                 2228.5 
                 2229.5 
               
               
                   
                 261-278 
                 AAPYEKEVPLSALTNILS 
                 204 
                 18 
                 DR4-2-65 
                 1916.2 
                 1917.4 
               
               
                 Cathepsin C 
                 151-167 
                 YDHNFVKAINADQKSWT 
                 118 
                 17 
                 DR4-2-70 
                 2037.2 
                 2039.6 
               
               
                 (Rat Homologue) 
                   
                 I 
                 119 
                   
                   
                 2035.3 
               
               
                   
                 151-166 
                 YDHNFVKAINADQKSW 
                 120 
                 16 
                 DR4-2-70 
                 1936.1 
                 1937.7 
               
               
                   
                   
                 I 
                 121 
                   
                   
                 1934.2 
               
               
                 Bovine Hemoglobin 
                 26-41 
                 AEALERMFLSFPTTKT 
                 205 
                 16 
                 DR4-2-78 
                 1842.1 
                 1836.1 
               
               
                 HLA-DQ3.2 β-chain 
                 24-38 
                 SPEDFVYQFKGMCYF 
                 206 
                 15 
                 DR4-2-78 
                 1861.1 
                 1861.7 
               
               
                 HLA-DR β-chain 
                 1-? 
                 GDTRPRFLEQVKHE . . . 
                 122 
                 14 
                 DR4-2-72 
                 1711.9 
               
               
                 IG Heavy Chain 
                 121-?  
                 GVYFYLQWGRSTLVSVS . . . 
                 123 
                 (? ) 
                 DR4-2-6 
                 ? 
                 ? 
               
               
                   
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 MANN/HLA-DR7 NATURALLY PROCESSED PEPTIDES 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 Pseudo HLA-A2 
                 105-124 
                 SDWRFLRGYHQYAYDGKDYI 
                 207 
                 20 
                 DR7-2-61 
                 2553.8 
                 2556.5 
               
               
                   
                 103-120 
                 VGSDWRFLRGYHQYAYDG 
                 1 
                 18 
                 DR7-2-63 
                 2190.4 
                 2194 
               
               
                   
                 103-117 
                 VGSDWRFLRGYHQYA 
                 2 
                 15 
                 DR7-2-63 
                 1855.0 
                 1860 
               
               
                   
                 104-117 
                 GSDWRFLRGYHQYA 
                 208 
                 14 
                 DR7-2-61 
                 1755.9 
                 1760.8 
               
               
                   
                 104-116 
                 GSDWRFLRGYHQY 
                 209 
                 13 
                 DR7-2-61 
                 1684.8 
                 1687.6 
               
               
                   
                 105-117 
                 SDWRFLRGYHQYA 
                 210 
                 13 
                 DR7-2-61 
                 1698.9 
                 1704.1 
               
               
                 HLA-A29 
                 234-253 
                 RPAGDGTFQKWASVVVPSGQ 
                 124 
                 20 
                 DR7-2-66 
                 2087.3 
                 2092 
               
               
                   
                 234-249 
                 RPAGDGTFQKWASVVV 
                 125 
                 16 
                 DR7-2-63 
                 1717 
                 1718 
               
               
                   
                 237-258 
                 GDGTFQKWASVVVPSGQEQRYT 
                 126 
                 22 
                 DR7-2-66 
                 2436 
                 2440 
               
               
                   
                 237-254 
                 GDGTFQKWASVVVPSGQE 
                 127 
                 18 
                 DR7-2-66 
                 1892.3 
                 1892 
               
               
                   
                 239-252 
                 GTFQKWASVVVPSG 
                 128 
                 14 
                 DR7-2-66 
                 1462 
                 1465 
               
               
                   
                 239-253 
                 GTFQKWASVVVPSGQ 
                 129 
                 15 
                 DR7-2-66 
                 1718 
                 1721 
               
               
                   
                 239-261 
                 GTFQKWASVVVPSGQEQRYTCHV 
                 130 
                 23 
                 DR7-2-66 
                 2603 
                 2606 
               
               
                 HLA-B44 
                 83-99 
                 RETQISKTNTQTYRENL 
                 211 
                 17 
                 DR7-2-35 
                 2082.3 
                 2086.1 
               
               
                   
                 83-98 
                 RETQISKTNTQTYREN 
                 212 
                 16 
                 DR7-2-35 
                 1969.1 
                 1971.1 
               
               
                   
                 83-97 
                 RETQISKTNTQTYRE 
                 213 
                 15 
                 DR7-2-35 
                 1855.0 
                 1857.3 
               
               
                 HLA-DR α-chain 
                 101-126 
                 RSNYTPITNPPEVTVLTNSPVELREP 
                 214 
                 26 
                 DR7-2-35 
                 2924.2 
                 2926.9 
               
               
                   
                 58-78 
                 GALANIAVDKANLEIMTKRSN 
                 131 
                 21 
                 DR7-2-66 
                 2229.5 
                 222I 
               
               
                   
                 182-200 
                 APSPLPETTENVVCALGLTV 
                 215 
                 20 
                 DR7-2-42 
                 1912.2 
                 1917.7 
               
               
                 HLA-DQ α-chain 
                 179-?  
                 SLQSPITVEWRAQSESAQSKMLSGIGGFVL 
                 216 
                 ? 
                 DR7-2-35 
                 ? 
                 ? 
               
               
                 4F2 Cell-surface 
                 318-338 
                 VTQYLNATGNRWCSWSLSQAR 
                 217 
                 21 
                 DR7-2-71 
                 2441.7 
                 2445.1 
               
               
                 antigen heavy chain 
                 318-334 
                 VTQYLNATGNRWCSWSL 
                 218 
                 17 
                 DR7-2-71 
                 1999.2 
                 2001.9 
               
               
                 LIF receptor 
                 854-866 
                 TSILCYRKREWIK 
                 219 
                 13 
                 DR7-2-35 
                 1696.0 
                 1700.8 
               
               
                 Ig kappa chain C reg. 
                 188-201 
                 KHKVYACEVTHQGL 
                 220 
                 14 
                 DR7-2-61 
                 1612.9 
                 1615.6 
               
               
                   
                 188-200 
                 KHKVYACEVTHQG 
                 221 
                 13 
                 DR7-2-61 
                 1498.7 
                 1501.0 
               
               
                 Invariant Chain 
                  99-120 
                 PKPPKPVSKMRMATPLLMQALP 
                 12 
                 22 
                 DR7-2-72 
                 2432.0 
                 2436.6 
               
               
                 (Ii) 
                 100-120 
                 KPPKPVSKMRMATPLLNQALP 
                 13 
                 21 
                 DR7-2-72 
                 2334.9 
                 2339.7 
               
               
                 K channel protein 
                 492-516 
                 GDMYPKTWSGMLVGALCALAGVLTI 
                 222 
                 25 
                 DR7-2-71 
                 2567.1 
                 2567.3 
               
               
                 Heat shock cognate 
                 38-54 
                 TPSYVAFTDTERLIGDA 
                 132 
                 17 
                 DR7-2-69 
                 1856.0 
                 1856.6 
               
               
                 71 KD protein 
                   
                   
                   
                 17 
                 DR7-2-72 
                 1856.0 
                 1857.0 
               
               
                   
                 38-52 
                 TPSYVAFTDTERLIG 
                 133 
                 15 
                 DR7-2-69 
                 1669.8 
                 1671.9 
               
               
                 Complement C9 
                 465-483 
                 APVLISQKLSPIYNLVPVK 
                 223 
                 19 
                 DR7-2-61 
                 2019.5 
                 2083.9 
               
               
                 Thromboxane-A 
                 406-420 
                 PAFRFTREAAQDCEV 
                 224 
                 15 
                 DR7-2-71 
                 1739.9 
                 1743.0 
               
               
                 synthase 
               
               
                 EBV major capsid prot 
                 1264-1282 
                 VPGLYSPCRAFFNKEELL 
                 225 
                 18 
                 DR7-2-54 
                 2082.4 
                 2081.2 
               
               
                   
                 1264-1277 
                 VPGLYSPCRAFFNK 
                 226 
                 14 
                 DR7-2-54 
                 1597.9 
                 1598.6 
               
               
                 Apolipoprotein B-100 
                 1586-1608 
                 KVDLTFSKQHALLCSDYQADYES 
                 227 
                 23 
                 DR7-2-54 
                 2660.9 
                 2662.5 
               
               
                   
                 1586-1600 
                 KVDLTFSKQHALLCS 
                 228 
                 15 
                 DR7-2-54 
                 1689.0 
                 1687.7 
               
               
                   
                 1942-1954 
                 FSHDYRGSTSHRL 
                 229 
                 13 
                 DR7-2-42 
                 1562.7 
                 1567.5 
               
               
                   
                 2077-2089 
                 LPKYFEKKRNTII 
                 230 
                 13 
                 DR7-2-61 
                 1650.0 
                 1653.8 
               
               
                   
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 23.1/HLA-DR8 NATURALLY PROCESSED PEPTIDES 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 HLA-DR α-chain 
                 158-180 
                 SETVFLPREDHLFRKFHYLPFLP 
                 231 
                 23 
                 DR8-3-59 
                 2889.3 
                 2889.0 
               
               
                   
                 182-198 
                 APSPLPETTENVVCALG 
                 232 
                 17 
                 DR8-3-41 
                 1697.9 
                 1704.3 
               
               
                 HLA-DR β-chain 
                 1-? 
                 GDTRPRFLEYSTGECYFFNGTERV 
                 233 
                 ? 
                 DR8-3-75 
                 — 
                 — 
               
               
                 HLA-DP β-chain 
                 80-92 
                 RHNYELDEAVTLQ 
                 234 
                 13 
                 DR8-3-76 
                 1587.7 
                 1591.3 
               
               
                 LAM Blast-1 with 
                  88-108 
                 DPQSGALYISKVQKEDNSTYI 
                 235 
                 21 
                 DR8-3-54 
                 2543.6 
                 2549.1 
               
               
                 N-acetyglucosamine 
                  92-108 
                 GALYISKVQKEDNSTYI 
                 236 
                 17 
                 DR8-3-52 
                 2116.1 
                 2118.0 
               
               
                   
                 129-146 
                 DPVPKPVIKIEKIEDMDD 
                 237 
                 18 
                 DR8-3-57 
                 2081.4 
                 2085.7 
               
               
                   
                 129-143 
                 DPVPKPVIKIEKIED 
                 238 
                 15 
                 DR8-3-57 
                 1720.0 
                 1724.9 
               
               
                 Ig kappa chain 
                 63-80 
                 FTFTISRLEPEDFAVYYC 
                 239 
                 18 
                 DR8-3-57 
                 2201.5 
                 2203.6 
               
               
                   
                 63-77 
                 FTFTISRLEPEDFAV 
                 240 
                 15 
                 DR8-3-57 
                 1772.0 
                 1777.0 
               
               
                 LAR protein 
                 1302-1316 
                 DPVEMRRLNYQTPG 
                 241 
                 14 
                 DR8-3-76 
                 1675.9 
                 1679.8 
               
               
                 LIF receptor 
                 709-726 
                 YQLLRSMIGYIEELAPIV 
                 242 
                 18 
                 DR8-3-66 
                 2108.5 
                 2112.0 
               
               
                 IFN-α receptor 
                 271-287 
                 GNHLYKWKQIPDCENVK 
                 243 
                 17 
                 DR8-3-66 
                 2072.4 
                 2075.1 
               
               
                 Interleukin-8 
                 169-188 
                 LPFFLFRQAYHPNNSSPVCY 
                 244 
                 20 
                 DR8-3-59 
                 2400.7 
                 2402.5 
               
               
                 receptor 
               
               
                 Metalloproteinase 
                 187-214 
                 QAKFFACIKRSDGSCAWYRGAAPPKQEF 
                 245 
                 28 
                 DR8-2-63 
                 3161.6 
                 3164.9 
               
               
                 inhibitor 2 
               
               
                   
                 187-205 
                 QAKFFACIKRSDGSCAWYR 
                 246 
                 19 
                 DR8-3-63 
                 2235.5 
                 2233.6 
               
               
                 Metalloproteinase 
                 101-118 
                 NRSEEFLIAGKLQDGLLH 
                 134 
                 18 
                 DR8-3-66 
                 2040.3 
                 2042.9 
               
               
                 inhibitor 1 
               
               
                   
                 101-117 
                 SEEFLIAGKLQDGLL 
                 135 
                 16 
                 DR8-3-70 
                 1789.0 
                 1799.9 
               
               
                   
                 103-117 
                 SEEFLIAGKLQDGLL 
                 247 
                 15 
                 DR8-3-72 
                 1632.9 
                 1646.0 
               
               
                   
                 101-112 
                 NRSEEFLIAGKL 
                 248 
                 12 
                 DR8-3-66 
                 1376.6 
                 1381.8 
               
               
                 Cathepsin E 
                  89-112 
                 QNFTVIFDTGSSNLWVPSVYCTSP 
                 249 
                 24 
                 DR8-3-59 
                 2662.9 
                 2664.4 
               
               
                 Cathepsin S 
                 189-205 
                 TAFQYIIDNKGIDSDAS 
                 68 
                 17 
                 DR8-3-63 
                 1857.9 
                 1857.1 
               
               
                 Cystatin SN 
                 41-58 
                 DEYYRRLLRVLRAREQIV 
                 250 
                 18 
                 DR8-3-63 
                 2448.7 
                 2348.0 
               
               
                 Tubulin α-chain 
                 207-223 
                 EAIYDICRRNLDIERPT 
                 251 
                 17 
                 DR8-3-63 
                 2077.3 
                 2078.3 
               
               
                   
                 207-219 
                 EAIYDICRRNLDI 
                 252 
                 13 
                 DR8-3-63 
                 1593.8 
                 1595.1 
               
               
                 Myosin β-heavy chain 
                 1027-1047 
                 HELEKIKKQVEQEKCEIQAAL 
                 253 
                 21 
                 DR8-3-59 
                 2493.9 
                 2494.0 
               
               
                 Ca release channel 
                 2614-2623 
                 RPSMLQHLLR 
                 254 
                 10 
                 DR8-3-68 
                 1250.5 
                 1254.8 
               
               
                 CD35 
                 359-380 
                 DDFMGQLLNGRVLFPVNLQLGA 
                 255 
                 22 
                 DR8-3-72 
                 2417.8 
                 2421.3 
               
               
                 CD75 
                 106-122 
                 IPRLQKIWKNYLSMNKY 
                 256 
                 17 
                 DR8-3-66 
                 2195.6 
                 2202.1 
               
               
                 c-myc transfor. prot. 
                 371-385 
                 KRSFFALRDQIPDL 
                 257 
                 14 
                 DR8-3-68 
                 1706.0 
                 1709.6 
               
               
                 K-ras trasnfor. prot. 
                 164-180 
                 RQYRLKKISKEEKTPGC 
                 258 
                 17 
                 DR8-3-54 
                 2064.4 
                 2066.5 
               
               
                 Calcitonin 
                 38-53 
                 EPFLYILGKSRVLEAQ 
                 69 
                 16 
                 DR8-3-78 
                 1863.2 
                 1848.4 
               
               
                 receptor (Hum?) 
               
               
                 α-ENOLASE (?) 
                 23-?  
                 AEVYHDVMSEFF . . . 
                 259 
                 ? 
                 DR8-3-54 
                 — 
                 — 
               
               
                 Plasminogen activator 
                 378-396 
                 DRPFLFVVRHNPTGTVLFM 
                 260 
                 19 
                 DR8-3-59 
                 2246.7 
                 2247.1 
               
               
                 inhibitor-1 
                 133-148 
                 MPHFFRLFRSTVKQVD 
                 261 
                 16 
                 DR8-3-70 
                 2008.4 
                 2116.4 
               
               
                 Apolipoprotein B-100 
                 1724-1743 
                 KNIFHFKVNQEGLKLSNDMM 
                 262 
                 20 
                 DR8-3-62 
                 2393.8 
                 2399.4 
               
               
                   
                 1724-1739 
                 KNIFHFKVNQEGLKLS 
                 263 
                 16 
                 DR8-3-57 
                 1902.2 
                 1903.7 
               
               
                   
                 1780-1799 
                 YKQTVSLDIQPYSLVTTLNS 
                 264 
                 20 
                 DR8-3-54 
                 2271.5 
                 2273.7 
               
               
                   
                 2646-2662 
                 STPEFTILNTLHIPSFT 
                 265 
                 17 
                 DR8-3-80 
                 1918.2 
                 1929.4 
               
               
                   
                 2647-2664 
                 TPEFTILNTLHIPSFTID 
                 266 
                 18 
                 DR8-3-80 
                 2059.3 
                 2073.5 
               
               
                   
                 2647-2662 
                 TPEFTILNTLHIPSFT 
                 267 
                 16 
                 DR8-3-80 
                 1831.1 
                 1841.6 
               
               
                   
                 2885-2900 
                 SNTKYFHKLNIPQLDF 
                 268 
                 16 
                 DR8-3-68 
                 1965.2 
                 1969.9 
               
               
                   
                 2072-2088 
                 LPFFKFLPKYFEKKRNT 
                 269 
                 17 
                 DR8-3-75 
                 2203.6 
                 2207.0 
               
               
                   
                 2072-2086 
                 LPFFKFLPKYFEKKR 
                 270 
                 15 
                 DR8-3-76 
                 1988.4 
                 1992.6 
               
               
                   
                 4022-4036 
                 WNFYYSPQSSPDKKL 
                 271 
                 15 
                 DR8-3-59 
                 1860.0 
                 1863.3 
               
               
                 Bovine Transferrin 
                 261-281 
                 DVIWELLNHAQEHFGKDKSKE 
                 272 
                 21 
                 DR8-3-76 
                 2523.8 
                 2524.9 
               
               
                   
                 261-275 
                 DVIWELLINHAQEHFG 
                 273 
                 15 
                 DR8-3-78 
                 1808.0 
                 1818.1 
               
               
                   
                 261-273 
                 DVIWELLNHAQEH 
                 196 
                 13 
                 DR8-3-73 
                 1603.8 
                 1608.8 
               
               
                 von Willebrand factor 
                 617-636 
                 IALLLMASQEPQRMSRNFVR 
                 190 
                 20 
                 DR8-3-59 
                 2360.8 
                 2359.7 
               
               
                   
                 617-630 
                 IALLLMASQEPQRM 
                 189 
                 14 
                 DR8-3-59 
                 1600.9 
                 1601.3 
               
               
                   
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 HOM2/HLA-DR1 NATURALLY PROCESSED PEPTIDES 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 FRACTION 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 Pseudo HLA-A2 
                 103-117 
                 VGSDWRFLRGYHQYA 
                 2 
                 15 
                 H2/DR1-1-64 
                 1855.0 
                 1854.4 
               
               
                   
                 104-117 
                 GSDWRFLRGYHQYA 
                 4 
                 14 
                 H2/DR1-1-63 
                 1755.3 
                 1755.2 
               
               
                 Invariant Chain 
                  98-121 
                 LPKPPKPVSKMRMATPLLMQALPM 
                 7 
                 24 
                 H2/DR1-1-77 
                 2676.4 
                 2675.9 
               
               
                 (Ii) 
                  99-122 
                 PKPPKPVSKMRMATPLLMQALPMG 
                 8 
                 24 
                 H2/DR1-1-72 
                 2620.2 
                 2619.7 
               
               
                   
                  98-120 
                 LPKPPKPVSKMRMATPLLMQALP 
                 9 
                 23 
                 H2/DR1-1-73 
                 2545.2 
                 2544.5 
               
               
                   
                  99-121 
                 PKPPKPVSKMRMATPLLMQALPM 
                 10 
                 23 
                 H2/DR1-1-75 
                 2563.2 
                 2562.3 
               
               
                   
                 100-121 
                 KPPKPVSKMRMATPLLMQALPM 
                 11 
                 22 
                 H2/DR1-1-75 
                 2466.1 
                 2465.8 
               
               
                   
                  99-120 
                 PKPPKPVSKMRMATPLLMQALP 
                 12 
                 22 
                 H2/DR1-1-72 
                 2432.0 
                 2431.7 
               
               
                   
                 100-120 
                 KPPKPVSKMRMATPLLMQALP 
                 13 
                 21 
                 H2/DR1-1-72 
                 2334.9 
                 2334.2 
               
               
                   
                   
                   
                   
                   
                   
                   
                 ESI-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 SUMMARY OF NATURALLY PROCESSSED PEPTIDES BOUND TO HLA-DR EXPRESSED IN NORMAL HUMAN SPLEEN 
               
             
          
           
               
                 PROTEIN SOURCE 
                 POSITION 
                 SEQUENCE 
                 SEQ ID NO. 
                 LENGTH 
                 MW 
                 MASS SPEC 
               
               
                   
               
             
          
           
               
                 HLA-DR α-chain 
                 71/133-156 
                 SETVFLPREDHLFRKFHYLPFLPS 
                 140 
                 24 
                 2976 
                 2982 
               
               
                   
                 71/136-156 
                 VFLPREDHLFRKFHYLPFLPS 
                 141 
                 21 
                 2659 
                 2666 
               
               
                   
                 71/136-155 
                 VFLPREDHLFRKFHYLPFLP 
                 142 
                 20 
                 2572 
                 2579 
               
               
                   
                 71/136-151 
                 VFLPREDHLFRKFHYL 
                 143 
                 16 
                 1118 
                 2126 
               
               
                 Calgranulin B 
                 33/25-33 
                 KLGHPDTLN 
                 144 
                 9 
                 994 
                  999 
               
               
                   
                 42/88-114 
                 WASHEKMHEGDEGPGHHHKPGLGEGTP 
                 145 
                 27 
                 2915 
                 2927 
               
               
                   
                 43/88-114 
                 WASHEKMHEGDEGPGHHHKPGLGEGTP 
                 146 
                 27 
                 2017 
                 2926 
               
               
                 HLA-B51 
                 42/104-121 
                 GPDGRLLRGHNQYDGK 
                 188 
                 16 
                 2017 
                 2023 
               
               
                 Kinase C ξ chain (rat) 
                 42/341-446 
                 TLPPFQPQITDDYGLD 
                 70 
                 16 
                 1704 
                 1705 
               
               
                 HLA-DR4 β chain 
                 45/129-144 
                 VRWFRNGQEEKTGVVS 
                 71 
                 16 
                 1892 
                 1894 
               
               
                   
                   
                   
                   
                   
                   
                 MALD-MS 
               
               
                   
               
             
          
         
       
     
     
       
         
           
             274 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              1
Val Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala Tyr
  1               5                  10                  15
Asp Gly
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              2
Val Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              3
Val Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr
  1               5                  10
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              4
Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              5
Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala
  1               5                  10
 
           
           
             
               25 
               amino acid 
                 
               linear 
             
              6
Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro
  1               5                  10                  15
Leu Leu Met Gln Ala Leu Pro Met Gly
             20                  25
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              7
Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro
  1               5                  10                  15
Leu Leu Met Gln Ala Leu Pro Met
             20
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              8
Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu
  1               5                  10                  15
Leu Met Gln Ala Leu Pro Met Gly
             20
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              9
Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro
  1               5                  10                  15
Leu Leu Met Gln Ala Leu Pro
             20
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              10
Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu
  1               5                  10                  15
Leu Met Gln Ala Leu Pro Met
             20
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              11
Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu
  1               5                  10                  15
Met Gln Ala Leu Pro Met
             20
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              12
Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu
  1               5                  10                  15
Leu Met Gln Ala Leu Pro
             20
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              13
Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu
  1               5                  10                  15
Met Gln Ala Leu Pro
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              14
Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu Met
  1               5                  10                  15
Gln Ala Leu Pro
             20
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              15
Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro Met
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              16
Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro
  1               5                  10
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              17
Ile Pro Ala Asp Leu Arg Ile Ile Ser Ala Asn Gly Cys Lys Val Asp
  1               5                  10                  15
Asn Ser
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              18
Arg Val Glu Tyr His Phe Leu Ser Pro Tyr Val Ser Pro Lys Glu Ser
  1               5                  10                  15
Pro
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              19
Tyr Lys His Thr Leu Asn Gln Ile Asp Ser Val Lys Val Trp Pro Arg
  1               5                  10                  15
Arg Pro Thr
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              20
Tyr Lys His Thr Leu Asn Gln Ile Asp Ser Val Lys Val Trp Pro Arg
  1               5                  10                  15
Arg Pro
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              21
Asp Val Gly Glu Tyr Arg Ala Val Thr Glu Leu Gly Arg Pro Asp Ala
  1               5                  10                  15
Glu Tyr Trp
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              22
Glu Pro Gly Glu Pro Glu Phe Lys Tyr Ile Gly Asn Met His Gly
  1               5                  10                  15
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              23
Tyr Lys His Thr Leu Asn Gln Ile Asp Ser Val Lys Val Trp Pro Arg
  1               5                  10                  15
Arg
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              24
Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr
  1               5                  10
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              25
Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr
  1               5                  10                  15
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              26
Ser Asp Leu Ser Phe Ser Lys Asp Trp Ser Phe Tyr Leu
  1               5                  10
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              27
Lys Val Phe Gly Arg Cys Glu Leu Ala Ala Ala Met Lys Arg His Gly
  1               5                  10                  15
Leu Asp
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              28
Arg Asn Arg Cys Lys Gly Thr Asp Val Gln Ala Trp Ile Arg Gly Cys
  1               5                  10                  15
Arg Leu
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              29
His Pro Pro His Ile Glu Ile Gln Met Leu Lys Asn Gly Lys Lys Ile
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              30
Asn Glu Leu Gly Arg Phe Lys His Thr Asp Ala Cys Cys Arg Thr His
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              31
Ser Lys Pro Lys Val Tyr Gln Trp Phe Asp Leu Arg Lys Tyr
  1               5                  10
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              32
Ala Thr Ser Thr Lys Lys Leu His Lys Glu Pro Ala Thr Leu Ile Lys
  1               5                  10                  15
Ala Ile Asp Gly
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              33
Pro Ala Thr Leu Ile Lys Ala Ile Asp Gly Asp Thr Val Lys Leu Met
  1               5                  10                  15
Tyr Lys Gly Gln
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              34
Asp Arg Val Lys Leu Met Tyr Lys Gly Gln Pro Met Thr Phe Arg Leu
  1               5                  10                  15
Leu Leu Val Asp
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              35
Val Ala Tyr Val Tyr Lys Pro Asn Asn Thr His Glu Gln His Leu Arg
  1               5                  10                  15
Lys Ser Glu Ala
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              36
Gln Lys Gln Glu Pro Ile Asp Lys Glu Leu Tyr Pro Leu Thr Ser Leu
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              37
Gly Ala Arg Ala Ser Val Leu Ser Gly Gly Glu Leu Asp Lys Trp Glu
  1               5                  10                  15
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              38
Arg Thr Leu Tyr Gln Asn Val Gly Thr Tyr Val Ser Val Gly Thr Ser
  1               5                  10                  15
Thr Leu Asn Lys
             20
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              39
Leu Lys Lys Leu Val Phe Gly Tyr Arg Lys Pro Leu Asp Asn Ile
  1               5                  10                  15
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              40
Lys His Ile Glu Gln Tyr Leu Lys Lys Ile Lys Asn Ser
  1               5                  10
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              41
Asp Val Phe Lys Glu Leu Lys Val His His Ala Asn Glu Asn Ile Phe
  1               5                  10                  15
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              42
Gly Asp Thr Arg Pro Arg Phe Leu Trp Gln Leu Lys Phe Glu Cys His
  1               5                  10                  15
Phe Phe Asn Gly
             20
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              43
Thr Glu Arg Val Arg Leu Leu Glu Arg Cys Ile Tyr Asn Gln Glu Glu
  1               5                  10                  15
Ser Val Arg Phe Asp Ser
             20
 
           
           
             
               25 
               amino acid 
                 
               linear 
             
              44
Asp Leu Leu Glu Gln Arg Arg Ala Ala Val Asp Thr Tyr Cys Arg His
  1               5                  10                  15
Asn Tyr Gly Val Gly Glu Ser Phe Thr
             20                  25
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              45
Lys Ala Glu Arg Ala Asp Leu Ile Ala Tyr Leu Lys Gln Ala Thr Ala
  1               5                  10                  15
Lys
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              46
Gly Arg Thr Gln Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile
  1               5                  10                  15
Val Thr Pro Arg Thr Pro Pro Pro
             20
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              47
Pro Leu Lys Ala Glu Ile Ala Gln Arg Leu Glu Asp Val
  1               5                  10
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              48
Arg Gln Ile Leu Gly Gln Leu Gln Pro Ser Leu Gln Thr Gly Ser Glu
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              49
Ile Gln Val Tyr Ser Arg His Pro Pro Glu Asn Gly Lys Pro Asn Ile
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              50
Ile Asn Thr Lys Cys Tyr Lys Leu Glu His Pro Val Thr Gly Cys Gly
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              51
Tyr Lys Leu Asn Phe Tyr Phe Asp Leu Leu Arg Ala Lys Leu
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              52
Ile Asp Thr Leu Lys Lys Asn Glu Asn Ile Lys Glu Leu
  1               5                  10
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              53
Asp Val Gly Glu Tyr Arg Ala Val Thr Glu Leu Gly Arg Pro Asp Ala
  1               5                  10                  15
Glu Tyr Trp Asn
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              54
Glu Arg Phe Ala Val Asn Pro Gly Leu Leu Glu Thr Ser Glu Gly Cys
  1               5                  10                  15
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              55
Asp Asn Tyr Arg Gly Tyr Ser Leu Gly Asn Trp Val Cys Ala Ala Lys
  1               5                  10                  15
Phe Glu Ser Asn Phe Thr Gln
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              56
Glu Ala Leu Val Arg Gln Gly Leu Ala Lys Val Ala Tyr Val Tyr Lys
  1               5                  10                  15
Pro Asn Asn Thr
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              57
Pro Ile Val Gln Asn Leu Gln Gly Gln Met Val His Gln Ala Ile Ser
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              58
Ser Ala Leu Ser Glu Gly Ala Thr Pro Gln Asp Leu Asn Thr Met Leu
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              59
Ser Phe Tyr Ile Leu Ala His Thr Glu Phe Thr Pro Thr Glu Thr Asp
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              60
Lys Met Tyr Phe Asn Leu Ile Asn Thr Lys Cys Tyr Lys Leu Glu His
  1               5                  10                  15
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              61
Val Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala Tyr
  1               5                  10                  15
Ala Asp
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              62
Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala Tyr Asp
  1               5                  10                  15
Gly
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              63
Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu Met
  1               5                  10                  15
Gln Ala Leu Pro
             20
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              64
Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro Met Gly
  1               5                  10                  15
Ala Leu Pro
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              65
Glu His His Ile Phe Leu Gly Ala Thr Asn Tyr Ile Tyr Val Leu Asn
  1               5                  10                  15
Glu Glu Asp Leu Gln Lys Val
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              66
Gln Glu Leu Lys Asn Lys Tyr Tyr Gln Val Pro Arg Lys Gly Ile Gln
  1               5                  10                  15
Ala
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              67
Ile Gln Asn Leu Ile Lys Glu Glu Ala Phe Leu Gly Ile Thr Asp Glu
  1               5                  10                  15
Lys Thr Glu Gly
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              68
Thr Ala Phe Gln Tyr Ile Ile Asp Asn Lys Gly Ile Asp Ser Asp Ala
  1               5                  10                  15
Ser
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              69
Glu Pro Phe Leu Tyr Ile Leu Gly Lys Ser Arg Val Leu Glu Ala Gln
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              70
Thr Leu Pro Pro Phe Gln Pro Gln Ile Thr Asp Asp Tyr Gly Leu Asp
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              71
Val Arg Trp Phe Arg Asn Gly Gln Glu Glu Lys Thr Gly Val Val Ser
  1               5                  10                  15
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              72
Arg Val Gln Pro Lys Val Thr Val Tyr Pro Ser Lys Thr Gln Pro Leu
  1               5                  10                  15
Gln His
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              73
Arg Val Gln Pro Lys Val Thr Val Tyr Pro Ser Lys Thr Gln Pro
  1               5                  10                  15
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              74
Asn Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr Leu Asn Lys Asn
  1               5                  10                  15
Ser Leu Lys
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              75
Ile Pro Asp Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr
  1               5                  10                  15
Leu Asn Lys
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              76
Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr Leu Asn Lys
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              77
Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr Leu Asn
  1               5                  10                  15
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              78
Tyr Ala Asn Ile Leu Leu Asp Arg Arg Val Pro Gln Thr Asp Met Thr
  1               5                  10                  15
Phe
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              79
Gly Asp Thr Arg Pro Arg Phe Leu Glu Tyr Ser Thr Ser Glu Cys His
  1               5                  10                  15
Phe Phe
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
  1               5                  10                  15
Pro Val Thr Lys Ser
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              81
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
  1               5                  10                  15
Pro Val Thr Lys
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              82
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
  1               5                  10                  15
Val Thr
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              83
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
  1               5                  10                  15
Pro
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              84
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
  1               5                  10                  15
Ser
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              85
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              86
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              87
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              88
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              89
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              90
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
  1               5                  10
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              91
Ala Pro Ser Pro Leu Pro Glu Thr Thr Glu Asn Val Val Cys Ala Leu
  1               5                  10                  15
Gly
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              92
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
Arg Met Glu Pro Arg
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              93
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
 Arg Met Glu Pro
              20
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              94
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
Arg Met Glu
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              95
Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln Arg Met
  1               5                  10                  15
Glu Pro Arg
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              96
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln Arg Met Glu
  1               5                  10                  15
Pro Arg Ala
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              97
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
Arg
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              98
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln Arg Met Glu
  1               5                  10                  15
Pro
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              99
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln Arg Met
  1               5                  10                  15
 
           
           
             
               12 
               amino acid 
                 
               linear 
             
              100
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser
  1               5                  10
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              101
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg Gly Glu Pro Arg Ala Pro
             20
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              102
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro Arg Gly Glu
  1               5                  10                  15
Pro Arg Ala Pro Trp Val
             20
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              103
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg Gly Glu Pro Arg
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              104
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg Gly Glu Pro
             20
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              105
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg Gly Glu
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              106
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg Gly
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              107
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro Arg Gly Glu
  1               5                  10                  15
Pro Arg
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              108
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro
  1               5                  10                  15
Arg
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              109
Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro Arg Gly
  1               5                  10                  15
Glu
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              110
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Pro Arg
  1               5                  10
 
           
           
             
               12 
               amino acid 
                 
               linear 
             
              111
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser
  1               5                  10
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              112
Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln Arg
  1               5                  10                  15
Lys Trp Glu Ala Ala
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              113
Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              114
Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              115
Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr
  1               5                  10                  15
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              116
Asp Leu Ser Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              117
Asp Leu Ser Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg Lys Trp Glu
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              118
Tyr Asp His Asn Phe Val Lys Ala Ile Asn Ala Asp Gln Lys Ser Trp
  1               5                  10                  15
Thr
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              119
Tyr Asp His Asn Phe Val Lys Ala Ile Asn Ala Asp Ile Lys Ser Trp
  1               5                  10                  15
Thr
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              120
Tyr Asp His Asn Phe Val Lys Ala Ile Asn Ala Asp Gln Lys Ser Trp
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              121
Tyr Asp His Asn Phe Val Lys Ala Ile Asn Ala Ile Gln Lys Ser Trp
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              122
Gly Asp Thr Arg Pro Arg Phe Leu Glu Gln Val Lys His Glu
  1               5                  10
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              123
Gly Val Tyr Phe Tyr Leu Gln Trp Gly Arg Ser Thr Leu Val Ser Val
  1               5                  10                  15
Ser
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              124
Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val
  1               5                  10                  15
Pro Ser Gly Gln
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              125
Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val
  1               5                  10                  15
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              126
Gly Asp Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val Pro Ser Gly
  1               5                  10                  15
Gln Glu Gln Arg Tyr Thr
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              127
Gly Asp Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val Pro Ser Gly
  1               5                  10                  15
Gln Glu
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              128
Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val Pro Ser Gly
  1               5                  10
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              129
Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val Pro Ser Gly Gln
  1               5                  10                  15
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              130
Gly Thr Phe Gln Lys Trp Ala Ser Val Val Val Pro Ser Gly Gln Glu
  1               5                  10                  15
Gln Arg Tyr Thr Cys His Val
             20
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              131
Gly Ala Leu Ala Asn Ile Ala Val Asp Lys Ala Asn Leu Glu Ile Met
  1               5                  10                  15
Thr Lys Arg Ser Asn
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              132
Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu Arg Leu Ile Gly Asp
  1               5                  10                  15
Ala
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              133
Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu Arg Leu Ile Gly
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              134
Arg Ser Glu Glu Phe Leu Ile Ala Gly Lys Leu Gln Asp Gly Leu Leu
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              135
Ser Glu Glu Phe Leu Ile Ala Gly Lys Leu Gln Asp Gly Leu Leu
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              136
Asp Val Ile Trp Glu Leu Leu Asn His Ala Gln Glu His Phe Gly
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              137
Glu Pro Phe Leu Tyr Ile Leu Gly Lys Ser Arg Val Leu Glu Ala Gln
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              138
Thr Ala Phe Gln Tyr Ile Ile Asp Asn Lys Gly Ile Asp Ser Asp
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              139
Thr Ala Phe Gln Tyr Ile Ile Asp Asn Lys Gly Ile Asp Ser
  1               5                  10
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              140
Ser Glu Thr Val Phe Leu Pro Arg Glu Asp His Leu Phe Arg Lys Phe
  1               5                  10                  15
His Tyr Leu Pro Phe Leu Pro Ser
             20
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              141
Val Phe Leu Pro Arg Glu Asp His Leu Phe Arg Lys Phe His Tyr Leu
  1               5                  10                  15
Pro Phe Leu Pro Ser
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              142
Val Phe Leu Pro Arg Glu Asp His Leu Phe Arg Lys Phe His Tyr Leu
  1               5                  10                  15
Pro Phe Leu Pro
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              143
Val Phe Leu Pro Arg Glu Asp His Leu Phe Arg Lys Phe His Tyr Leu
  1               5                  10                  15
 
           
           
             
               26 
               amino acid 
                 
               linear 
             
              144
Lys Leu Gly His Pro Asp Thr Leu Asn Gln Gly Glu Phe Lys Glu Leu
  1               5                  10                  15
Val Arg Lys Asp Leu Gln Asn Phe Leu Lys
             20                  25
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              145
Lys Leu Gly His Pro Asp Thr Leu Asn Gln Gly Glu Phe Lys Glu Leu
  1               5                  10                  15
Val Arg Lys Asp Leu Gln Asn Phe
             20
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              146
Lys Leu Gly His Pro Asp Thr Leu Asn Gln Gly Glu Phe Lys
  1               5                  10
 
           
           
             
               123 
               nucleic acid 
               single 
               linear 
             
              147
ATG GCC ATA AGT GGA GTC CCT GTG CTA GGA TTT TTC ATC ATA GCT GTG  48
Met Ala Ile Ser Gly Val Pro Val Leu Gly Phe Phe Ile Ile Ala Val
1               5                   10                  15
CTG ATG AGC GCT CAG GAA TCA TGG GCT AAG ATG CGC ATG GCC ACC CCG  96
Leu Met Ser Ala Gln Glu Ser Trp Ala Lys Met Arg Met Ala Thr Pro
            20                  25                  30
CTG CTG ATG CAG GCG CTG CCC ATG TAA                             123
Leu Leu Met Gln Ala Leu Pro Met
        35                  40
 
           
           
             
               150 
               nucleic acid 
               single 
               linear 
             
              148
ATG GCC ATA AGT GGA GTC CCT GTG CTA GGA TTT TTC ATC ATA GCT GTG  48
Met Ala Ile Ser Gly Val Pro Val Leu Gly Phe Phe Ile Ile Ala Val
1               5                   10                  15
CTG ATG AGC GCT CAG GAA TCA TGG GCT CTT CCC AAG CCT CCC AAG CCT  96
Leu Met Ser Ala Gln Glu Ser Trp Ala Leu Pro Lys Pro Pro Lys Pro
            20                  25                  30
GTG AGC AAG ATG CGC ATG GCC ACC CCG CTG CTG ATG CAG GCG CTG CCC 144
Val Ser Lys Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro
        35                  40                  45
ATG TAA                                                         150
Met
 
           
           
             
               10 
               amino acid 
                 
               linear 
             
              149
Thr Gln Phe Val Arg Phe Asp Ser Asp Ala
  1               5                  10
 
           
           
             
               10 
               amino acid 
                 
               linear 
             
              150
Asp Trp Arg Phe Leu Arg Gly Tyr His Gln
  1               5                  10
 
           
           
             
               10 
               amino acid 
                 
               linear 
             
              151
Arg Met Ala Thr Pro Leu Leu Met Gln Ala
  1               5                  10
 
           
           
             
               4 
               amino acid 
                 
               linear 
             
              152
Lys Asp Glu Leu
  1
 
           
           
             
               5 
               amino acid 
                 
               linear 
             
              153
Lys Phe Glu Arg Gln
  1               5
 
           
           
             
               5 
               amino acid 
                 
               linear 
             
              154
Gln Arg Glu Phe Lys
  1               5
 
           
           
             
               25 
               amino acid 
                 
               linear 
             
              155
Met Ala Ile Ser Gly Val Pro Val Leu Gly Phe Phe Ile Ile Ala Val
  1               5                  10                  15
Leu Met Ser Ala Gln Glu Ser Trp Ala
             20                  25
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              156
Met Arg Met Ala Thr Pro Leu Leu Met Gln Ala Leu Pro Met
  1               5                  10
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              157
Met Pro Arg Ser Arg Ala Leu Ile Leu Gly Val Leu Ala Leu Thr Thr
  1               5                  10                  15
Met Leu Ser Leu Cys Gly Gly
             20
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              158
Asn Ile Val Ile Lys Arg Ser Asn Ser Thr Ala Ala Thr Asn Glu Val
  1               5                  10                  15
Pro Glu Val Thr Val Phe Ser
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              159
Asn Ile Val Ile Lys Arg Ser Asn Ser Thr Ala Ala Thr Asn Glu Val
  1               5                  10                  15
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              160
Ser Asp Val Gly Val Tyr Arg Ala Val Thr Pro Gln Gly Arg Pro Asp
  1               5                  10                  15
Ala Glu
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              161
Asp Val Gly Val Tyr Arg Ala Val Thr Pro Gln Gly Arg Pro Asp Ala
  1               5                  10                  15
Glu
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              162
Asp Val Gly Val Tyr Arg Ala Val Thr Pro Gln Gly Arg Pro Asp
  1               5                  10                  15
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              163
Ala Pro Ser Pro Leu Pro Glu Thr Thr Glu Asn Val Val
  1               5                  10
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              164
Ala Pro Ser Pro Leu Pro Glu Thr Thr Glu Asn Val Val Cys Ala Leu
  1               5                  10                  15
Gly
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              165
Phe Pro Lys Ser Leu His Thr Tyr Ala Asn Ile Leu Leu Asp Arg Arg
  1               5                  10                  15
Val Pro Gln Thr Asp
             20
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              166
Phe Pro Lys Ser Leu His Thr Tyr Ala Asn Ile Leu Leu Asp Arg Arg
  1               5                  10                  15
Val Pro Gln
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              167
Asp Gly Ile Leu Tyr Tyr Tyr Gln Ser Gly Gly Arg Leu Arg Arg Pro
  1               5                  10                  15
Val Asn
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              168
Asp Gly Ile Leu Tyr Tyr Tyr Gln Ser Gly Gly Arg Leu Arg Arg Pro
  1               5                  10                  15
Val
 
           
           
             
               30 
               amino acid 
                 
               linear 
             
              169
Leu Ser Pro Ile His Ile Ala Leu Asn Phe Ser Leu Asp Pro Gln Ala
  1               5                  10                  15
Pro Val Asp Ser His Gly Leu Arg Pro Ala Leu His Tyr Gln
             20                  25                  30
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              170
Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn Arg
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              171
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              172
Pro Pro Glu Val Thr Val Leu Thr Asn Ser Pro Val Glu Leu Arg Glu
  1               5                  10                  15
Pro Asn Val
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              173
Pro Pro Glu Val Thr Val Leu Thr Asn Ser Pro Val Glu Leu Arg Glu
  1               5                  10                  15
Pro Asn
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              174
Val Phe Leu Leu Leu Leu Ala Asp Lys Val Pro Glu Thr Ser Leu Ser
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              175
Thr Phe Asp Glu Ile Ala Ser Gly Phe Arg Gln Gly Gly Ala Ser Gln
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              176
Tyr Gly Tyr Thr Ser Tyr Asp Thr Phe Ser Trp Ala Phe Leu
  1               5                  10
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              177
Ala Thr Lys Tyr Gly Asn Met Thr Glu Asp His Val Met His Leu Leu
  1               5                  10                  15
Gln Asn Ala
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              178
Gly Gln Val Lys Lys Asn Asn His Gln Glu Asp Lys Ile Glu
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              179
Leu Asn Lys Ile Leu Leu Asp Glu Gln Ala Gln Trp Lys
  1               5                  10
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              180
Gly Pro Pro Lys Leu Asp Ile Arg Lys Glu Glu Lys Gln Ile Met Ile
  1               5                  10                  15
Asp Ile Phe His
             20
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              181
Gly Pro Pro Lys Leu Asp Ile Arg Lys Glu Glu Lys Gln Ile Met Ile
  1               5                  10                  15
Asp Ile Phe His Pro
             20
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              182
Ser Pro Leu Gln Ala Leu Asp Phe Phe Gly Asn Gly Pro Pro Val Asn
  1               5                  10                  15
Tyr Lys Thr Gly Asn Leu
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              183
Ser Pro Leu Gln Ala Leu Asp Phe Phe Gly Asn Gly Pro Pro Val Asn
  1               5                  10                  15
Tyr Lys Thr Gly
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              184
Gly Lys Phe Ala Ile Arg Pro Asp Lys Lys Ser Asn Pro Ile Ile Arg
  1               5                  10                  15
Thr Val
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              185
Thr Gly His Gly Ala Arg Thr Ser Thr Glu Pro Thr Thr Asp Tyr
  1               5                  10                  15
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              186
Lys Glu Leu Lys Arg Gln Tyr Glu Lys Lys Leu Arg Gln
  1               5                  10
 
           
           
             
               12 
               amino acid 
                 
               linear 
             
              187
Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala
  1               5                  10
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              188
Gly Pro Asp Gly Arg Leu Leu Arg Gly His Asn Gln Tyr Asp Gly Lys
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              189
Ile Ala Leu Leu Leu Met Ala Ser Gln Glu Pro Gln Arg Met
  1               5                  10
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              190
Ile Ala Leu Leu Leu Met Ala Ser Gln Glu Pro Gln Arg Met Ser Arg
  1               5                  10                  15
Asn Phe Val Arg
             20
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              191
Ile Pro Asp Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr
  1               5                  10                  15
Leu Asn Lys Asn
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              192
Ile Pro Asp Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr
  1               5                  10                  15
Leu Asn
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              193
Ile Pro Asp Asn Leu Phe Leu Lys Ser Asp Gly Arg Ile Lys Tyr Thr
  1               5                  10                  15
Leu
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              194
Val Thr Thr Leu Asn Ser Asp Leu Lys Tyr Asn Ala Leu Asp Leu Thr
  1               5                  10                  15
Asn
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              195
Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln
  1               5                  10                  15
Arg Met Glu Pro Arg Ala Pro
             20
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              196
Asp Val Ile Trp Glu Leu Leu Asn His Ala Gln Glu His
  1               5                  10
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              197
Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg Lys Trp
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              198
Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln Arg
  1               5                  10                  15
Lys Trp
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              199
Asp Leu Ser Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg Lys Trp Glu Ala Ala
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              200
Asp Leu Ser Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Thr Gln
  1               5                  10                  15
Arg Lys
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              201
Gly Ser Leu Phe Val Tyr Asn Ile Thr Thr Asn Lys Tyr Lys Ala Phe
  1               5                  10                  15
Leu Asp Lys Gln
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              202
Gly Ser Leu Phe Val Tyr Asn Ile Thr Thr Asn Lys Tyr Lys Ala Phe
  1               5                  10                  15
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              203
Ala Ala Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile
  1               5                  10                  15
Leu Ser Ala Gln Leu
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              204
Ala Ala Pro Tyr Glu Lys Glu Val Pro Leu Ser Ala Leu Thr Asn Ile
  1               5                  10                  15
Leu Ser
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              205
Ala Glu Ala Leu Glu Arg Met Phe Leu Ser Phe Pro Thr Thr Lys Thr
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              206
Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Met Cys Tyr Phe
  1               5                  10                  15
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              207
Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala Tyr Asp Gly
  1               5                  10                  15
Lys Asp Tyr Ile
             20
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              208
Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              209
Gly Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              210
Ser Asp Trp Arg Phe Leu Arg Gly Tyr His Gln Tyr Ala
  1               5                  10
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              211
Arg Glu Thr Gln Ile Ser Lys Thr Asn Thr Gln Thr Tyr Arg Glu Asn
  1               5                  10                  15
Leu
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              212
Arg Glu Thr Gln Ile Ser Lys Thr Asn Thr Gln Thr Tyr Arg Glu Asn
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              213
Arg Glu Thr Gln Ile Ser Lys Thr Asn Thr Gln Thr Tyr Arg Glu
  1               5                  10                  15
 
           
           
             
               26 
               amino acid 
                 
               linear 
             
              214
Arg Ser Asn Tyr Thr Pro Ile Thr Asn Pro Pro Glu Val Thr Val Leu
  1               5                  10                  15
Thr Asn Ser Pro Val Glu Leu Arg Glu Pro
             20                  25
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              215
Ala Pro Ser Pro Leu Pro Glu Thr Thr Glu Asn Val Val Cys Ala Leu
  1               5                  10                  15
Gly Leu Thr Val
             20
 
           
           
             
               30 
               amino acid 
                 
               linear 
             
              216
Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser Glu Ser
  1               5                  10                  15
Ala Gln Ser Lys Met Leu Ser Gly Ile Gly Gly Phe Val Leu
             20                  25                  30
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              217
Val Thr Gln Tyr Leu Asn Ala Thr Gly Asn Arg Trp Cys Ser Trp Ser
  1               5                  10                  15
Leu Ser Gln Ala Arg
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              218
Val Thr Gln Tyr Leu Asn Ala Thr Gly Asn Arg Trp Cys Ser Trp Ser
  1               5                  10                  15
Leu
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              219
Thr Ser Ile Leu Cys Tyr Arg Lys Arg Glu Trp Ile Lys
  1               5                  10
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              220
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu
  1               5                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              221
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
  1               5                  10
 
           
           
             
               25 
               amino acid 
                 
               linear 
             
              222
Gly Asp Met Tyr Pro Lys Thr Trp Ser Gly Met Leu Val Gly Ala Leu
  1               5                  10                  15
Cys Ala Leu Ala Gly Val Leu Thr Ile
             20                  25
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              223
Ala Pro Val Leu Ile Ser Gln Lys Leu Ser Pro Ile Tyr Asn Leu Val
  1               5                  10                  15
Pro Val Lys
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              224
Pro Ala Phe Arg Phe Thr Arg Glu Ala Ala Gln Asp Cys Glu Val
  1               5                  10                  15
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              225
Val Pro Gly Leu Tyr Ser Pro Cys Arg Ala Phe Phe Asn Lys Glu Glu
  1               5                  10                  15
Leu Leu
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              226
Val Pro Gly Leu Tyr Ser Pro Cys Arg Ala Phe Phe Asn Lys
  1               5                  10
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              227
Lys Val Asp Leu Thr Phe Ser Lys Gln His Ala Leu Leu Cys Ser Asp
  1               5                  10                  15
Tyr Gln Ala Asp Tyr Glu Ser
             20
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              228
Lys Val Asp Leu Thr Phe Ser Lys Gln His Ala Leu Leu Cys Ser
  1               5                  10                  15
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              229
Phe Ser His Asp Tyr Arg Gly Ser Thr Ser His Arg Leu
  1               2                  10
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              230
Leu Pro Lys Tyr Phe Glu Lys Lys Arg Asn Thr Ile Ile
  1               5                  10
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              231
Ser Glu Thr Val Phe Leu Pro Arg Glu Asp His Leu Phe Arg Lys Phe
  1               5                  10                  15
His Tyr Leu Pro Phe Leu Pro
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              232
Ala Pro Ser Pro Leu Pro Glu Glu Thr Thr Glu Asn Val Val Cys Ala
  1               5                  10                  15
Leu Gly
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              233
Gly Asp Thr Arg Pro Arg Phe Leu Glu Tyr Ser Thr Gly Glu Cys Tyr
  1               5                  10                  15
Phe Phe Asn Gly Thr Glu Arg Val
             20
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              234
Arg His Asn Tyr Glu Leu Asp Glu Ala Val Thr Leu Gln
  1               5                  10
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              235
Asp Pro Gln Ser Gly Ala Leu Tyr Ile Ser Lys Val Gln Lys Glu Asp
  1               5                  10                  15
Asn Ser Thr Tyr Ile
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              236
Gly Ala Leu Tyr Ile Ser Lys Val Gln Lys Glu Asp Asn Ser Thr Tyr
  1               5                  10                  15
Ile
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              237
Asp Pro Val Pro Lys Pro Val Ile Lys Ile Glu Lys Ile Glu Asp Met
  1               5                  10                  15
Asp Asp
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              238
Asp Pro Val Pro Lys Pro Val Ile Lys Ile Glu Lys Ile Glu Asp
  1               5                  10                  15
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              239
Phe Thr Phe Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr
  1               5                  10                  15
Tyr Cys
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              240
Phe Thr Phe Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val
  1               5                  10                  15
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              241
Asp Pro Val Glu Met Arg Arg Leu Asn Tyr Gln Thr Pro Gly
  1               5                  10
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              242
Tyr Gln Leu Leu Arg Ser Met Ile Gly Tyr Ile Glu Glu Leu Ala Pro
  1               5                  10                  15
Ile Val
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              243
Gly Asn His Leu Tyr Lys Trp Lys Gln Ile Pro Asp Cys Glu Asn Val
  1               5                  10                  15
Lys
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              244
Leu Pro Phe Phe Leu Phe Arg Gln Ala Tyr His Pro Asn Asn Ser Ser
  1               5                  10                  15
Pro Val Cys Tyr
             20
 
           
           
             
               28 
               amino acid 
                 
               linear 
             
              245
Gln Ala Lys Phe Phe Ala Cys Ile Lys Arg Ser Asp Gly Ser Cys Ala
  1               5                  10                  15
Trp Tyr Arg Gly Ala Ala Pro Pro Lys Gln Glu Phe
             20                  25
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              246
Gln Ala Lys Phe Phe Ala Cys Ile Lys Arg Ser Asp Gly Ser Cys Ala
  1               5                  10                  15
Trp Tyr Arg
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              247
Ser Glu Glu Phe Leu Ile Ala Gly Lys Leu Gln Asp Gly Leu Leu
  1               5                  10                  15
 
           
           
             
               12 
               amino acid 
                 
               linear 
             
              248
Asn Arg Ser Glu Glu Phe Leu Ile Ala Gly Lys Leu
  1               5                  10
 
           
           
             
               24 
               amino acid 
                 
               linear 
             
              249
Gln Asn Phe Thr Val Ile Phe Asp Thr Gly Ser Ser Asn Leu Trp Val
  1               5                  10                  15
Pro Ser Val Tyr Cys Thr Ser Pro
             20
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              250
Asp Glu Tyr Tyr Arg Arg Leu Leu Arg Val Leu Arg Ala Arg Glu Gln
  1               5                  10                  15
Ile Val
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              251
Glu Ala Ile Tyr Asp Ile Cys Arg Arg Asn Leu Asp Ile Glu Arg Pro
  1               5                  10                  15
Thr
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              252
Glu Ala Ile Tyr Asp Ile Cys Arg Arg Asn Leu Asp Ile
  1               5                  10
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              253
His Glu Leu Glu Lys Ile Lys Lys Gln Val Glu Gln Glu Lys Cys Glu
  1               5                  10                  15
Ile Gln Ala Ala Leu
             20
 
           
           
             
               10 
               amino acid 
                 
               linear 
             
              254
Arg Pro Ser Met Leu Gln His Leu Leu Arg
  1               5                  10
 
           
           
             
               22 
               amino acid 
                 
               linear 
             
              255
Asp Asp Phe Met Gly Gln Leu Leu Asn Gly Arg Val Leu Phe Pro Val
  1               5                  10                  15
Asn Leu Gln Leu Gly Ala
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              256
Ile Pro Arg Leu Gln Lys Ile Trp Lys Asn Tyr Leu Ser Met Asn Lys
  1               5                  10                  15
Tyr
 
           
           
             
               14 
               amino acid 
                 
               linear 
             
              257
Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Asp Leu
  1               5                  10
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              258
Arg Gln Tyr Arg Leu Lys Lys Ile Ser Lys Glu Glu Lys Thr Pro Gly
  1               5                  10                  15
Cys
 
           
           
             
               13 
               amino acid 
                 
               linear 
             
              259
Ala Glu Val Tyr His Asp Val Ala Ala Ser Glu Phe Phe
  1               5                  10
 
           
           
             
               19 
               amino acid 
                 
               linear 
             
              260
Asp Arg Pro Phe Leu Phe Val Val Arg His Asn Pro Thr Gly Thr Val
  1               5                  10                  15
Leu Phe Met
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              261
Met Pro His Phe Phe Arg Leu Phe Arg Ser Thr Val Lys Gln Val Asp
  1               5                  10                  15
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              262
Lys Asn Ile Phe His Phe Lys Val Asn Gln Glu Gly Leu Lys Leu Ser
  1               5                  10                  15
Asn Asp Met Met
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              263
Lys Asn Ile Phe His Phe Lys Val Asn Gln Glu Gly Leu Lys Leu Ser
  1               5                  10                  15
 
           
           
             
               20 
               amino acid 
                 
               linear 
             
              264
Tyr Lys Gln Thr Val Ser Leu Asp Ile Gln Pro Tyr Ser Leu Val Thr
  1               5                  10                  15
Thr Leu Asn Ser
             20
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              265
Ser Thr Pro Glu Phe Thr Ile Leu Asn Thr Leu His Ile Pro Ser Phe
  1               5                  10                  15
Thr
 
           
           
             
               18 
               amino acid 
                 
               linear 
             
              266
Thr Pro Glu Phe Thr Ile Leu Asn Thr Leu His Ile Pro Ser Phe Thr
  1               5                  10                  15
Ile Asp
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              267
Thr Pro Glu Phe Thr Ile Leu Asn Thr Leu His Ile Pro Ser Phe Thr
  1               5                  10                  15
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              268
Ser Asn Thr Lys Tyr Phe His Lys Leu Asn Ile Pro Gln Leu Asp Phe
  1               5                  10                  15
 
           
           
             
               17 
               amino acid 
                 
               linear 
             
              269
Leu Pro Phe Phe Lys Phe Leu Pro Lys Tyr Phe Glu Lys Lys Arg Asn
  1               5                  10                  15
Thr
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              270
Leu Pro Phe Phe Lys Phe Leu Pro Lys Tyr Phe Glu Lys Lys Arg
  1               5                  10                  15
 
           
           
             
               15 
               amino acid 
                 
               linear 
             
              271
Trp Asn Phe Tyr Tyr Ser Pro Gln Ser Ser Pro Asp Lys Lys Leu
  1               5                  10                  15
 
           
           
             
               21 
               amino acid 
                 
               linear 
             
              272
Asp Val Ile Trp Glu Leu Leu Asn His Ala Gln Glu His Phe Gly Lys
  1               5                  10                  15
Asp Lys Ser Lys Glu
             20
 
           
           
             
               16 
               amino acid 
                 
               linear 
             
              273
Asp Val Ile Trp Glu Leu Leu Ile Asn His Ala Gln Glu His Phe Gly
  1               5                  10                  15
 
           
           
             
               23 
               amino acid 
                 
               linear 
             
              274
Met Pro Arg Ser Arg Ala Leu Ile Leu Gly Val Leu Ala Leu Thr Thr
  1               5                  10                  15
Met Leu Ser Leu Cys Gly Gly
             20