AAMP-1, A PROTEIN WITH LOCAL HOMOLOGIES TO HIV-1 ENV AND NEF PROTEINS

Abstract: The present invention relates, in general, to AAMP-1. In particular, the present invention relates to a DNA segment encoding AAMP-1; polypeptides encoded by said DNA segment; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing an AAMP-1 polypeptide; antibodies specific to AAMP-1; and a method of measuring the amount of AAMP-1 in a sample.

Description: AAMP-l, A PROTEIN WITH LOCAL HOMOLOGIES TO HIV-1 ENV AND NEF PROTEINS. BACKGROUND 07 THE ZMVBHTZOK Field of the Invention The present invention relates, in general, to AAMP-l. In particular, the present invention relates to a DNA segment encoding AAMP- 1; polypeptides encoded by the DNA segment; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing AAMP-l; antibodies specific to AAMP-l; and a method of measuring the amount of AAMP-l in a sample. Background Information The major histocompatibility complex class II proteins have recently been found to contain local homologies to the HIV-1 envelope protein (H. Golding et al., J. EXP. Med. 167, 914 (1988); H. Golding et al., J. Clin. Invest. 83, 1430 (1989); J. A. T. Young, Nature 332, 215 (1988)). Such homologous regions may serve as targets for antibodies generated to HIV-l proteins and thus compromise the immune system in AIDS. Golding et al. (J. EXP. Med. 167, 914 (1988)) have identified a common epitope located in the carboxy terminus of the HIV-l gp41-envelope protein and the amino terminal portion of the beta chain of all human HLA class II antigens. Although the epitope is small, 5 consecutive identities or similarities, they found that it is an effective example of "molecular mimicry n in that monoclonal antibodies raised against synthetic peptides from each protein react interchangeably with native HIV-l envelope and MHC class II molecules. One third of HIV-l positive individuals were shown to have serum antibodies directed against peptides derived from HIV-l envelope protein, the homologous peptide from the MHC class XI molecules, and native MHC class II molecules (H. Golding et al. J. EXP. Med. 167, 914 (1988)). Two other regions of the HLA class II beta chain and another immune related protein, interleukin-2, also show limited homology to HIV-l (J. A. T. Young, Nature 333, 215 (1988)).; M.A. Vega et al. Nature 345, 26 1990).; W. E. Reiher III, et al. Proc. Natl. Acad. Sci. USA 83. 9188 (1986)). An important consideration in HIV-l vaccine development is the potential existence of additional host cell surface proteins with ho ologies to HIV-l that may crossreact with antibodies directed against its peptides. The present invention relates to the protein AAMP-l which has immunoglobulin (Ig) type domains that contain strong local homologies to conserved regions of the HIV-l envelope and nef proteins. SUMMARY OF THE INVENTION It is a general object of this invention to provide AAMP-l. It is a specific object of this invention to provide a DNA segment which encodes AAMP-l, or segment thereof. It is a further object of the invention to provide a polypeptide corresponding to a AAMP- 1 gene, or fragment thereof. It is another object of the invention to provide a recombinant DNA molecule comprising a vector and a DNA segment encoding a AAMP-l gene. It is a further object of the invention to provide a cell that contains the above- described recombinant molecule. It is another object of the invention to provide a method of producing a polypeptide encoding a AAMP-l gene, or segment thereof. It is a further object of the invention to provide antibodies having binding affinity for AAMP-l, or a unique portion thereof. It is a further object of the invention to provide a method of measuring the amount of AAMP-l in a sample. It is another object of the invention to provide a therapeutic modality comprising the above-described polypeptides in an amount effective to elicit protective antibodies, block harmful auto-antibodies, or compete for HIV binding to body cells in a patient to the AIDS virus and a pharmaceutically acceptable diluent, carrier, or excipient. It is a further object of the invention to provide a method of preventing AIDS in a patient. Further objects and advantages of the present invention will be clear from the description that follows. BRIEF DESCRIPTION 07 THE DRAWINGS Figure 1. Nucleotide sequence of human A2058 melanoma cell AAMP-l cDNA isolated from a lambda gtll expression library with its predicted amino acid sequence. The phage insert, AAMP-l, was subcloned into Bluescript plasmid (Stratagene) for production of double stranded cDNA for sequencing using the dideoxynucleotide termination method (F. Sanger et al. Proc. Natl. Acad. Sci. USA 74, 5463 (1977)) with Sequenase 2.0 (U.S. Biochemical) . Nucleotide residues are numbered beginning at the 5' end. Amino acid sequence numbering begins with the first methionine (underlined with "=") of the open reading frame. The amino terminal acidic region, aa26-85, is underlined with " ". Amino acid region, 76- 346, comprised of potential immunoglobulin-like domains (A. F. Williams and A. N. Barclay, Ann R?Vτ lFF"n?l- 6, 381 (1988) ; A. F. Williams and A. N. Barclay, in Immunoglobulin Genes. T. Honjo et al. Eds. (Academic Press Limited, San Diego, CA, 1989), pp. 361-387)) is underlined with secondary structure predications of beta strands, "****", and beta turns, n »» n , based on the method of Chou and Fasman fAdvances in Enz. 47, 45 (1978)). Cysteine pairs, 139 t 208, and 265 & 326, predicted by immunoglobulin domain homology to most likely form disulfide bonds are marked "< >". The potential transmembrane region, aa374-399, is underlined, " ". The potential protein kinase C phosphorylation site at serine #408 is underlined with "$$$$" (The protein kinase C phosphorylation site consensus sequence (Ser/Thr- Xaa-Lys/Arg) where Xaa is usually an uncharged residue [J. R. Woodgett, K. L. Gould, T. Hunter, Eur. J. Biochem. 161, 177 (1986)], is also found at threonines #238, 291, and 357)). The polyadenylation site at nucleic acid residues, 1723-1729 is in parentheses "()". Figure 2. Northern blot of human melanoma A2058 cells probed with AAMP-l cDNA. A single 1.6Kb band is seen on blots of total cytoplas ic (Lane 1) and polyadenylate-enriched (Lane 2) A2058 RNA. Total cytoplasmic RNA, 41 micrograms (μg) , was isolated from 6 million cells lysed in Nonidet P-40 (0.65%), separated into an aqueous phase in the presence of 7M urea, 1% sodium dodecyl sulfate, Tris buffer, NaCl, and EDTA, followed by phenol/chloroform extraction. RNA, 2.2 μg, enriched for messenger RNA, was isolated from 16 million cells with a Fast Track Kit Version 2.1 (Invitrogen Corp. San Diego, CA) . RNA was denatured in formaldehyde, electrophoresed in a 1% agarose/formaldehyde gel, transferred to Schlelcher & Schnell Nytran nylon membrane and crosslinked with ultraviolet light. The 1766 bp AAMP-l cDNA was labeled with (alpha-"P) dCTP (NEN Research Products, Boston, MA) using random priming. Hybridization overnight at 65*C was performed according to Church and Gilbert (Proc. Natl. Acad. Sci 81, 1991 (1984)). Figure 3. Northern blot of AAMP-l expression in human T-cell activation. Hours refer to time in culture. A: AAMP-l single 1.6Kb message. B: Beta-2 micro-globulin standard. Lanes 1-3: Non-stimulated human CD4+ T cells (Human peripheral blood mononuclear cells from normal donors were separated by Ficoll-Hypaque density-gradient centrifugation. Unstimulated CD4+lymphocytes were obtained by rigorous immunomagnetic negative selection with Advanced Magnetic Particles (Advanced Magnetic, Cambridge, MA) or Dynabeads (Dynal Inc., Fort Lee, NJ) both bound to goat anti-mouse lgG. Negative selection was performed as described (K. J. Horgan and S. Shaw, Current Protocols in Immunology. J. E. Eoligan et al, Eds. (Wiley Interscience, new York, 1991), p. 7.44.1.) using a cocktail of monoclonal antibodies, mAbs, consisting of anti-HLA class II mAb (IVA12) , CD20 mAb (1F5) , CD16 mAb (3G8) , CDllb mAb (NIHllb-1), CD14 mAb (MMA) , CD8 mAb (B9.8), and mAb against glycophorin (10F7) . Purity of the isolated cells was greater than 98%. The selected CD4+ T-cellε were free of monocytes based on the lack of proliferative response to optimal concentrations (1/200 dilution) of Phytohe agglutinin (M form) (PHA) (GIBCO, Grand Island, NY)), lanes 1 and 2 at 0 and 24 hours, respectively, without mitogen stimulation, and lane 3 after 12 hours in the presence of the protein synthesis inhibitor, cycloheximide, which has been frequently observed to stabilize certain mRNA species (K. Kelly et al. P. Leder, Cell 35, 603 (1983)). Lanes 4-8: CD4+T cells activated ". (T-cell activation assays were performed using >■ standard techniques. Briefly, 10 million purified CD4+ T-cellε were cultured in 35 mm flat bottom wells for various time periods in culture medium (RPM1 1640 (Hazelton Biologies Inc. Lenexa, KS) supplemented with 20 mM glutamine (Hazelton) , 10% heat inactivated fetal calf serum (Biofluids, Rockville, MD) , 100 IU/ml of penicillin, and 100 μg/ l streptomycin) , either unstimulated or stimulated with antibodies bound to the wells. T- cell stimulatory conditions were as described (G. A. van Seventer et al. Eur. J. jp ..nr>7 - 21, 1711 (1991)). Monoclonal antibodies were immobilized on the plastic of the well by dilution in phosphate buffered saline (PBS) and overnight incubation at 4*C, followed by washing with PBS. The CD3 mAb, OKT3, and the CD2 mAb, 95-5-49, were applied at 1 μg and 10 μg, purified j/ml --- respectively, all in a volume of 3 milliliters per well. Monoclonal antibodies were used as purified immunoglobulin derived from ascites fluid; CD2 mAb (directed against the Tll.l epitope): 95-5-49, lgGl (hybridoma kindly provided by Dr. R. R. Quinones, George Washington University, Washington, DC); CD3 mAb 0KT3, lgG2a (ATCC, Rockville, MD) . Cycloheximide, when present, was used at a concentration of 10 μg/ml. Lanes 4,5,6,7 and 8, represent the time points at 1, 2, 4, 16, and 24 hours, respectively. RNA samples were prepared from CD4+ T cells by the guanidinium isothiocyanate-cesiu chloride method of Maniatis et al. (T. E. Maniatis, E. F. Fritsch, J. Sambrook, Molecular Clonin g : A aboratory MfllWal (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, ed. 2, 1989), pp. 7.18-7.22.)). Ten micrograms of total RNA (each lane) electrophoresed in a formaldehyde/0.8% agarose gel was transferred to nitrocellulose and hybridized overnight, consecutively, at 42*C to the (alpha- M P) dCTP labeled, random primed probes, AAMP-l and beta-2 microglobulin. DETAILED DESCRIPTION 07 THE INVENTION In one embodiment, the present invention relates to a DNA segment coding for a polypeptide comprising an amino acid sequence corresponding to AAMP-l, or at least 5 contiguous amino acids thereof. In one preferred embodiment, the DNA segment comprises the sequence shown in SEQ ID NO:l, allelic or species variation thereof, or at least 15 contiguous nucleotides thereof (preferably, at least 20, 30, 40, or 50 contiguous nucleotides thereof) . In a further preferred embodiment, the DNA segment encodes the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof (preferably, at least 5, 10, 15, 20, 30 or 50 contiguous amino acids thereof) . In a further embodiment, the present invention relates to a polypeptide free of proteins with which it is naturally associated or a polypeptide bound to a solid support and comprising an amino acid sequence corresponding to AAMP-l, or at least 5 contiguous amino acids thereof (preferably, at least 5, 10, 15, 20, 30 or 50 contiguous amino acids thereof) . In one preferred embodiment, the polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2, or allelic or species variation thereof equivalent thereto (for example, immunologically or functionally, equivalent thereto) , or at least 5 contiguous amino acids thereof (preferably, at least 5, 10, 15, 20, 30 or 50 contiguous amino acids thereof) . In another embodiment, the present invention relates to a recombinant DNA molecule comprising a vector (for example plasmid or viral vector) and a DNA segment (as described above) coding for a polypeptide corresponding to AAMP-l; as described above. In a preferred embodiment, the encoding segment is present in the vector operably linked to a promoter. In a further embodiment, the present invention relates to a cell containing the above described recombinant DNA molecule. Suitable host cells include procaryotes (such as bacteria, including E_j. coli) and both lower eucaryotes (for example yeast) and higher eucaryotes (for example, mammalian cells) . Introduction of the recombinant molecule into the cell can be effected using methods known in the art. In another embodiment, the present invention relates to a method of producing a polypeptide having an amino acid sequence corresponding to AAMP-l comprising culturing the above-described cell under conditions such that the DNA segment is expressed and the polypeptide thereby produced and isolating the polypeptide. In yet another embodiment, the present invention relates to an antibody having binding affinity for AAMP-l, or a unique portion thereof. In one preferred embodiment, AAMP-l comprises the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof (preferably, at least 5, 10, 15, 20, 30 or 50 contiguous amino acids thereof) . In one preferred embodiment, the antibody is 1AA3. Antibodies (monoclonal or polyclonal) can be raised to AAMP-l, or unique portions thereof, in its naturally occuring form and in its recombinant form. Binding fragments of such antibodies are also within the scope of the invention. AAMP-l may be joined to other materials, particularly polypeptides, as fused or covalently joined polypeptides to be used as immunogens. AAMP-l or its fragments may be fused or covalently linked to a variety of immunogens, such as keyhole limpet hemocyanin, bovine serum albumin, tetanus toxoid, etc. See for example. Microbiology, Hoeber Medical Division (Harper and Row, 1969) , Landsteiner, Specificity of Serological Reactions (Dover Publications, New York, 1962) and Williams et al., Methods in Immunology and Immunochemistry, Vol. 1 (Academic Press, New York, 1967) , for descriptions of methods of preparing polyclonal antisera. A typical method involves hyperimmunization of an animal with an antigen. The blood of the animal is then collected shortly after the repeated immunizations and the gamma globulin is isolated. In some instances, it is desirable to prepare monoclonal antibodies from various mammalian hosts. Description of techniques for preparing such monoclonal antibodies may be found in Stites et al., editors, Basic and Clinical Immunology, (Lange Medical Publications, Los Altos, CA, Fourth edition) and references cited therein, and in particular in Kohler and Milstein in Nature 256:495-497 (1975), which discusses one method of generating monoclonal antibodies. In another embodiment, the present invention relates to a hybridoma which produces a monoclonal antibody or binding fragment thereof having binding affinity for AAMP-l. In one preferred embodiment, AAMP-l has the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof (preferably, at least 5, 10, 15, 20, 30 or 50 contiguous amino acids thereof) . In another preferred embodiment, the hybridoma comprises 1AA3. In yet another embodiment, the present invention relates to a diagnostic kit comprising: i) at least one of the above-described monoclonal antibodies, and ii) a conjugate comprising a binding partner of said monoclonal antibody and a label. In a further embodiment, the present invention relates to a diagnostic kit comprising a conjugate comprising: i) at least one of the above-described monoclonal antibodies, and ii) a label. In a further embodiment, the present invention relates to a method of measuring the amount of AAMP-l in a sample, comprising contacting the sample with the above-described antibodies and measuring the amount of immunocomplexes formed between the antibodies and any AAMP-l in the sample. Methods of measuring the amount of immunocomplexes formed can be those well known in the art, such as RIA, ELISA, and direct and indirect immunoassays. In another embodiment, the present invention relates to a therapeutic agent suitable for use in protecting against HIV infection or treating inflammatory immune or neoplastic disorders comprising the above-identified polypeptides in a quantity selected depending on the route of administration. Although subcutaneous or intramuscular routes of administration are preferred, the above described polypeptides could also be administered by an intraperitoneal or intravenous route. One skilled in the art will appreciate that the amounts to be administered for any particular treatment protocol can be readily determined. Suitable amounts might be expected to fall within the range of 1-50 micromoles. In another embodiment, the present invention relates to a method of using the above described polypeptide to prevent AIDS. One skilled in the art will appreciate that the amounts to be administered for any particular treatment protocol can readily be determined. The present invention is described in further detail in the following non-limiting Examples. EXAMPLES The following protocols and experimental details are referenced in the Examples that follow: Cells. Human peripheral blood mononuclear cells (PBMC) from normal donors were separated by Ficoll-Hypaque density-gradient centrifugation. Resting CD4+ T lymphocytes were subsequently obtained by rigorous immunomagnetic negative selection with Advanced Magnetic Particles (Advanced Magnetic, Cambridge, MA) or Dynabeads (Dynal Inc. , Fort Lee, NJ) both bound to goat anti-mouse IgG. Negative selection was performed as described (Horgan, K.J. and Shaw, S., Immuno¬ magnetic negative selection of lymphocyte subsets in Coligan, J.E. et al. (Eds.) Current Protocols in Immunology, Wiley Interscience, New York (1991) p. 7.4.1.) using a cocktail of mAbs consisting of anti-HLA class II mAb (IVA12) , CD20 mAb (1F5) , CD16 mAb (3G8) CDllb mAb (NIHllb-1) , CD14 mAb (MMA) , CD8 mAb (B9.8), and mAb against glycophorin (10F7) . Purity of the isolated cells was more than 98%. The selected CD4+ T-cells were free of monocytes based on the criterion that there be no proliferative response to optimal concentrations (1/200 dilution) of Phytohemagglutinin (M form) (PHA) (GIBCO, Grand Island, NY) (Davis, L., and P.E. Lipsky (1986) J. Immunol. 136:3588). T-cell activation assays. T-cell activation assays were performed using standard techniques. Briefly 10x10' purified CD4+ T-cells were cultured in 35mm flat bottom wells for various time periods in culture medium [RPMJ 1640 (Hazleton Biologies Inc. Lenexa, KS) supplemented with 20 mM glutamine (Hazleton), 10% heat inactivated FCS (Biofluids, Rockville, MD) , 100 IU/ml of penicillin, and 100 μg/ml streptomycin) ] , either unstimulated or stimulated with antibodies bound to the wells. T- cell stimulatory conditions were as described (van Seventer, G.A. et al. (1991) Eur. J. Immuol. 21:1711). mAbs were immobilized on the plastic of the well by dilution in PBS and overnight incubation at 4*C, followed by washing with PBS. The CD3 mAb OKT3 and the CD2 mAb 95-5-49 were applied at 1 μg and 10 μg purified Ig/ml respectively, all in a volume of 3 ml/well. Monoclonal following antibodies were used as purified immunoglobulin derived from ascites fluid; CD2 mAb (directed against the Tll.l epitope): 95-5-49, IgGl (hybridoma kindly provided by Dr. R.R. Quinones, George Washington University, Washington, D.C.); CD3 mAb OKT3, IgG2a (ATCC, Rockville, MD) . Cycloheximide, when present, was used at a concentration of 10 μg/ml. CD4+ T cell RNA preparation. RNA samples were prepared from CD4+ T cells by the guanidinium isothiocyanate-CsCl method of Maniatis et al. (Maniatis, T.E. et al. (1989) Molecular cloning: a laboratory manual. 2nd Edition. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.). 10 μg of total RNA was resolved for each condition on a formaldehyde 0.8% agarose gel, transferred to nitrocellulose, and hybridized at 42'C to M P- labeled purified AAMP-l cDNA insert prepared by random priming. Antibody Preparation. The adaptive passive transfer technique in Balb/c mice utilizing whole cells from the human melanoma A2058 cell line as antigen was used to generate hybridomas with myeloma cells. Selection of the 1AA3 clone was based on its inhibition of motility when assayed in modified Boyden chambers described previously (Stracke, M.L. et al. (1987) Biochem. Biophys. Res. Comm. 146, 339-345) using gelatin coated filters and various chemoattractants (collagen type IV, laminin, autocrine otility factor, fibronectin, and insulin-like growth factor I. The clone 1AA3 was recloned by limiting dilution to produce the 1AA3AA clone. cDNA Library and Screening. The human melanoma A2058 cDNA expression library was constructed in the lambda gtll vector by Clontech Laboratories, Inc. Y1090 Escherichia coli infected by the phage were plated and blotted onto nitrocellulose filters (Schleicher & Schnell) for immunoassay with 1AA3AA antibody. Reactive plaques were detected using peroxidase-coupled antibody specific for mouse IgG. Northern Blotting. Preparation of A2058 human melanoma RNA enriched for messenger RNA was isolated with a Fast Track Kit Version 2.1 (Invitrogen Corp) . Total cytoplasmic RNA was isolated according to a published method (Gough, N.M. (1988) Anal. Biochem. 173, 93-95) by suspending 4ml cells on ice with 0.8ml chilled Solution A (10 mM Tris Cl, pH 7.5, 0.15 MNaCl, 1.5 mM Mgclj, and 0.65% Nonidet P-40) . The supernate, obtained after vortexing and centrifuging (800 x G, 5 in, 4*C) was mixed with 0.8 ml Solution B (7M Urea, 1% SDS, 0.35 M NaCl, 10 mM EDTA, pH 8.0, and 10 mM Tris Cl, pH 7.5) and 1.6 ml. Solution C (phenol: chloroform: isoamyl alcohol (50:50:1). RNA was removed in the aqueous phase and ethanol precipitated. RNA was denatured in formaldehyde, separated on a 1% agarose/formaldehyde gel (Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY) and transferred overnight to S&S Nytran (Schleicher & Schnell) and crosslinked to it with ultraviolet light in the Stratalinker apparatus (Stratagene) . The 1766 bp cDNA insert labeled with (α-"P)dCTP (NEN Research Products) with the Random Primer DNA Labeling System (Bethesda Research Laboratories, Life Technologies, Inc.) was used as probe. Northern blots for total A2058 melanoma cytoplasmic RNA and RNA enriched for messenger RNA were performed with the Church protocol (Church, G. and Gilbert, W. (1984) Proc. Natl. Acad. Sci 81, 1991) . The filter was washed at 65*C for 20 minutes with wash buffer (1% sodium dodecyl sulfate, 40 mM NaH.PO., 1 mM EDTA) three times and then autoradiographed at -70'C. DNA Sequencing. Positive phage inserts were subcloned into Bluescript plasmid (phagemid) (Stratagene) for production of DNA for sequencing. Double-stranded cDNA was sequenced using the dideoxynucleotide chain termination method with Seguenase (United States Biochemical) . Sequence obtained with the SK primer (Stratagene) specific for the adjacent Bluescript vector region was determined first. Subsequent sequencing utilized primers prepared on site based on previously obtained sequence for both strands completely. Sequence Data Analysis. GenBank (Intel!igenetics, Inc.) was searched with the program and further analyses of the sequence was accomplished with RAOARGOS, PESTFIND, PROSITE, AACLUST, KERMIT NALIGN, PALIGN, REPEATS, SEQIN, TRANSL, AND DIAPRO programs in PC/Gene (Intelligenetics, Inc.). The NBRF protein sequence data base from the Protein Identification Resource National Biomedical Research Foundation (NBRF) was searched with the PQS, XQS, and NEW programs and other programs were used for sequence analyses. In the ALIGN program, sequences are matched with a bias and gap penalty, scored in a matrix, scrambled and rescored many times to yield a mean best random score and standard deviation (SD) . The score for the real sequences is expressed as the number of SD units away from the random mean score (Dayhoff, M.O. et al. (1983) Meth. Enzvm. 91, 524-545). All of our alignments were done with the Mutation Data Matrix (250 PAMs) , md, a bias of 6, a gap penalty of 6 and 150 random runs (Williams, A.F. and Barclay, A.N. (1988) Ann. Rev- yunni-no] . 6- 381-405). EXAMPLE 1 Characterization of AAMP-l AAMP-l Antibodies. The monoclonal antibody produced against AAMP-l is of the IgG-I subtype. It cryoprecipitates and loses activity with freezing and purification methods that require precipitation. Initial results indicated that this antibody inhibited adhesion and motility of A2058 melanoma cells in chemoattractant assays performed with the modified Boyden chamber. However, the inhibition occurred in an all or none fashion without a reliable dose response curve and steric hindrance due to self aggregation of the antibody cannot be ruled out at this time. Characterization of the Proteins Identified bv 1AA3AA Antibody. A2058 melanoma cell surface immunofluorescent staining has been seen with . 1AA3AA. It identifies a protein on A2058 whole cell lysate immunoblots with a molecular weight of approximately 95kD that shows an apparent slight increase with reduction to 105kD. The betagalactosidase fusion protein shows positive staining with the 1AA3AA antibody on immunoblots. The predicted AAMP-l protein is described below. Its molecular weight and glycosylation potential are not consistent with the protein identified by 1AA3AA described above. Isolation of 1AA3AA Positive cDNA Clones. Initial screening of phage plaques yielded three positive clones similar in size, identified as 1AA34A, 1AA335A, and AAMP-l. They all cross hybridized with each other on dot blots. AAMP-l- was slightly larger (less than lObp different) and was chosen for sequencing. Northern Blot of A2058 Melanoma Total Cytoplasmic and Polyadenylate Enriched RNA. When all three positive clones were used to probe a blot of total cytoplasmic A2058 RNA they hybridized with only one band. A single band at 1.6kb is seen on a blot of both total cytoplasmic and polyadenylate enriched A2058 RNA probed with AAMP-l in Figure l. Nucleotide Sequence. The AAMP-l cDNA has 1766bp with the longest open reading frame (1245bp) occurring in the second reading frame of the sequence (Figure 2; SEQ ID NO:l). 67% of the sequence excluding the poly A tail is involved with repeats that include 7 or more nucleotides each. The largest direct repeat is A G G A G G A A G A G at nucleotides #201 and #1685. Its sequence overlaps with that of a ten member repeat at nucleotides #197 and #1428. Another ten member direct repeat occurs at positions #948 and #1508 and a third 10 member repeat is at #1111 and #1171. Several palindromes exist in the sequence. The longest palindrome G G G T T C T A G A A C C C occurs at nucleotide #228. Ten member palindromes also occur at nucleotides #1149 and #1340. Eight member palindromes are present at nucleotides #228, #1119, #1515, and #1710. The last 25 nucleotides of the 1766bp sequence comprise the polyadenylated nucleotide tail and the consensus sequence A A T A A A A that commonly precedes a poly A tail is present at nucleotide #1723. Predicted Amino Acid Seouence. The 1245bp open reading frame in AAMP-l cDNA predicts a protein with a molecular weight of at least 44 kilodaltons when the first methionine in the sequence (coded by the twelfth codon) is assumed to be the initiating methionine (Figure 2; SEQ ID NO:2) . The predicted protein contains multiple immunoglobulin-like domains qualifying it as a member of the immunoglobulin (Ig) superfamily. It contains two potential transmembrane regions and several serine/threonine phosphorylation sites. An acidic amino terminal region is also present. Immunoglobulin Superfamily Homoloov. Nucleic acid similarity with CD4, an immunoglobulin superfamily member, prompted a search for Ig domains in AAMP- 1 protein. Fourteen cysteines are present with eight present on the amino terminal side of the potential transmembrane regions (TMR) . Seven cysteine pairs have 57 - 78 intervening amino acids. These sizes are consistent with those found in Ig domains. Immunoglobulin V type domains usually have 65 - 75 intervening amino acids and C type domains have 55 - 60. Additional cysteine pairs with 43 and 44 intervening amino acids were also evaluated to find all possible domains with significant homology to the Ig domains of the superfamily members. Several of - It - these potential domains are overlapping so that fewer domains (1-3) are actually possible. Significant homology was interpreted as alignments yielding ALIGN scores of greater than 3.0 SD using the ALIGN program available in the NBRF data base package (Protein Identification Resource (1991) Protein Sequence Database, National Biomedical Research Foundation, Washington, DC) . The appropriate parameters for detecting immunoglobulin-like domains as specified by Williams and Barclay were used and are listed in the Methods. Scores of 3.1, 4.3, and 5.2 SD units indicate chance probabilities of 10 ~3 , 10"', and 10 '7 , respectively, for aligned sequences to show similarities unless they are related. Significant alignment scores were found for both V and C2 type overlapping immunoglobulin domains in AAMP-l. Cysteine #265 shows significant homology either as the first cysteine forming a disulfide bond or as the second cysteine in an overlapping domain. When V type immunoglobulin domains are matched against the putative AAMP-l domain including cysteines #265 and #326 forming the predicted disulfide bond, significant ALIGN scores can be obtained with immunoglobulin domains from several Ig superfamily members. Typical immunoglobulin domains are defined as beginning 20 amino acids before the first cysteine and ending 20 amino acids after the second cysteine of the disulfide bond. A truncated version of the putative domain, aa252- 326, also matched against several V type domains resulting in enhanced ALIGN scores compared to those associated with the complete domain and revealed additional significant homologies when matched with several more members of the Ig superfamily, thus, increasing the probability that this is a Ig domain. Four Ig domains showed significant homology with the overlapping AAMP-l region involving cysteines #208 and #265, aal88-285. A larger, overlapping and inclusive region, involving cysteines #208 and #282, region aal88- 302, yielded less significant ALIGN scores overall. Another region of AAMP-l involving cysteine #139 shows Ig domain homology of a lesser degree. Putative Ig domains with C#139 as either the first or the second cysteine forming a disulfide bond show significant Tg domain homology. The putative domain of AAMP-l including the aall9-228 region utilizes cysteine #139 as the first cysteine in the predicted disulfide bond. Its alignment is significant with at least three other Ig domains at the present time listed in Table I and the half domain matches with additional half domains (myelin basic glycoprotein, MAG(IV), 3.95SD; NCAM (V), 3.64SD; and polylgR(II) , 3.11). The other possible Ig domain involves cysteine #139 as the second cysteine in a disulfide bond and includes the region, aa79-159. Other shorter significant alignments were found while searching the Ig superfamily and its relatives for homology with AAMP-l. One involves a 17 amino acid region of CD4 (RWHUT4) which is slightly-proximal to its human immunodeficiency virus I (HIV-I) binding site matching the region including AAMP-l cysteine #265 with an ALIGN score of 5.03SD. There are two regions in the HIV-l gpl20 envelope protein that are similar and one region in the HIV-l nef protein that is similar to AAMP-l. Other Ig superfamily members found in the PIR database whose Ig domains were included in the searches for significant homology with AAMP-l showed less homology and are listed as follows: MUC18 (PIR3:A34507) domains I-V, human melanoma cell surface glycoprotein; CD8 (RWHUT8) V region domain, human T cell surface glycoprotein CD8 precursor; Po protein (PIR3:JQ0622) V region domain, rat peripheral myelin protein 0 precursor; Ig lambda chain C region; Ig heavy chain V (G1HUNM) V region domain, human immunoglobulin heavy chain V-II region; CEA (PIR3:A36319) domains I, IV, and V, human carcinoembryonic antigen (clone cosCEAl) ; TcR-beta V (RWHUVY) V region domain, human T-cell receptor beta chain precursor V region; Thy-1 (TDHU) V region domain, human Thy- 1 membrane glycoprotein precursor; MRC OX-2 (TDRTOX ) V and C region domains, rat OX-2 membrane glycoprotein precursor; CD3 epsilon (PIR2:A25769) C region domain, human T cell surface glycoprotein CD3 epsilon chain; ICAM (PIR2:S00573) domains I-V, human intercellular adhesion molecule 1; Ig kappa C (K3HU) C region domain, human immunoglobulin kappa chain C region; alpha-1-beta glycoprotein (OMHU1B) domains I and II, human alph-1-beta-glycoprotein; MHC-II beta (HLHU3D) non Ig type and C region domains, human major histocompatibility antigen, class II, beta chain; amalgam (PIR3:A31923) domains I-III, Drosophila melanogaster; platelet derived growth factor receptor (PDGF-R) domain (V) , and human lymphocyte surface antigen precursor CDW4 (PIR3:A32376) . AAMP-l Internal Homology. Significant internal homology is also seen within AAMP-l, predominantly involving the putative Ig domains. The region, aall9-169, including cysteine #139 shows homology with at least three other regions containing cysteines that may or may not be involved with disulfide bonds. These are listed: Another significant alignment is obtained by matching the putative domains, region aal88- 285 with region 76-159 yielding an ALIGN score of 6.63 SD. Secondary Structure Predictions for Putative Immunoglobulin Domains in AAMP-l. The AAMP-l region, aa76-346, encompassing all of the potential Ig domains has predicted secondary structure characteristics consistent with what is found in Ig domains. The Ig fold consists of two beta sheets each containing 3-4 anti-parallel beta strands (or sheets) . The region, aa245-346, contains 4-8 beta sheets separated by 10 potential beta turns. The region, aall9-228, contains 8-9 beta sheets separated by 10 potential beta turns. The other two regions, aa76-159 and aa!88-228 are similar. These predictions are from the PIR CHOFAS-Protein Secondary Structure Prediction Program. Potential Transmembrane Regions. There are two potential transmembrane regions predicted according to the method of Rao and Argos. Their method uses a conformational preference parameter for membrane-buried helices called the "buried- helix parameter" based on hydration potential, free energy of transfer from aqueous helix to nonpolar helix, polarity, bulk conformational preference, and turn conformational preference expressed as a sum of values for each amino acid. (Rao) One 24 amino acid region that met their criteria included aa323-347 with aspartic acid #338 having the highest "buried helix parameter" of 1.216. The other potential transmembrane region, aa374-399, has a peak value of 1.181 and contains no charged residues. Comparisons were made with other Ig superfamily members' transmembrane regions (CD2, TcRa alpha, CD4, Poly Ig R, MAG, and NCAM) . No significant alignments were found for the entire predicted transmembrane regions but an alignment score of 3.15 SD was obtained for a 19 amino acid region (aa379-397) of the second AAMP-l transmembrane region mentioned above with CD4's transmembrane region. Potential Phosphorylation Sites. On the amino terminal side of the AAMP-l TMRs there are five sites that have the consensus pattern for potential casein kinase II phosphorylation sites, (S,T)-x-x-(E,D) . These involve serines at positions #3, #122, and #308 and threonines at positions #109 and #165. Four potential protein kinase C phosphorylation sites are also present with the consensus pattern of (S,T)-x-(R,K) . These include two threonines at positions #238 and #291 on the amino terminal side of the TMR and a threόnine at position #357 and a serine at position $308 on the carboxy terminal side of the TMR. * * * * * All publications mentioned hereinabove are hereby incorporated in their entirety by reference. While the foregoing invention has been described in some detail for purposes of clarity and understanding, it will be appreciated by one skilled in the art from a reading of this disclosure that various changes in form and detail can be made without departing from the true scope of the invention and appended claims. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Beckner, Marie E. Liotta, Lance A. (ii) TITLE OF INVENTION: AAMP-l (iii) NUMBER OF SEQUENCES: 2 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: CUSHMAN, DARBY & CUSHMAN (B) STREET: Eleventh Floor, 1615 L. Street, N. . (C) CITY: Washington (D) STATE: D.C. (E) COUNTRY: USA (F) ZIP: 20036 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (202)861-3000 (B) TELEFAX: (202) 822-0944 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1766 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 35..1279 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GGGCCAGAGA AGTGGAATCC GCCGCTTGCG CCGC ATG GAG TCC GAA TCG GAA Met Glu Ser Glu Ser Glu 1 5 AGC GGG GCT GCT GCT GAC ACC CCC CCA CTG GAG ACC CTA AGC TTC CAT 1 Ser Gly Ala Ala Ala Asp Thr Pro Pro Leu Glu Thr Leu Ser Phe His 10 15 20 GGT GAT GAA GAG ATT ATC GAG GTG GTA GAA CTT GAT CCC GGT CCG CCG 1 Gly Asp Glu Glu lie lie Glu Val Val Glu Leu Asp Pro Gly Pro Pro 25 30 35 GAC CCA GAT GAC CTG GCC CAG GAG ATG GAA GAT GTG GAC TTT GAG GAA 1 Asp Pro Asp Asp Leu Ala Gin Glu Met Glu Asp Val Asp Phe Glu Glu 40 45 50 GAA GAG GAG GAA GAG GGC AAC GAA GAG GGC TGG GTT CTA GAA CCC CAG 2 Glu Glu Glu Glu Glu Gly Asn Glu Glu Gly Trp Val Leu Glu Pro Gin 55 60 65 70 GAA GGG GTG GTC GGC AGC ATG GAG GGC CCC GAC GAT AGC GAG GTC ACC Glu Gly Val Val Gly Ser Met Glu Gly Pro Asp Asp Ser Glu Val Thr 75 80 85 TTT GCA TTG CAC TCA GCA TCT GTG TTT TGT GTG AGC CTG GAC CCC AAG Phe Ala Leu His Ser Ala Ser Val Phe Cys Val Ser Leu Asp Pro Lys 90 95 100 ACC AAT ACC TTG GCA GTG ACC GGG GGT GAA GAT GAC AAA GCC TTC GTA Thr Asn Thr Leu Ala Val Thr Gly Gly Glu Asp Asp Lys Ala Phe Val 105 110 115 TGG CGG CTC AGC GAT GGG GAG CTG CTC TTT GAG TGT GCA GGC CAT AAA Trp Arg Leu Ser Asp Gly Glu Leu Leu Phe Glu Cys Ala Gly His Lys 120 125 130 GAC TCT GTG ACT TGT GCT GGT TTC AGC CAT GAC TCC ACT CTA GTG GCC Asp Ser Val Thr Cys Ala Gly Phe Ser His Asp Ser Thr Leu Val Ala 135 140 145 150 ACA GGG GAC ATG AGT GGC CTC TTG AAA GTG TGG CAG GTG GAC ACT AAG Thr Gly Asp Met Ser Gly Leu Leu Lys Val Trp Gin Val Asp Thr Lys 155 160 165 GAG GAG GTC TGG TCC TTT GAA GCG GGA GAC CTG GAG TGG ATG GAG TGG Glu Glu Val Trp Ser Phe Glu Ala Gly Asp Leu Glu Trp Met Glu Trp 170 175 180 CAT CCT CGG GCA CCT GTC CTG TTG GCG GGC ACA GCT GAC GGC AAC ACC His Pro Arg Ala Pro Val Leu Leu Ala Gly Thr Ala Asp Gly Asn Thr 185 190 195 TGG ATG TGG AAA GTC CCG AAT GGT GAC TGC AAG ACC TTC CAG GGT CCC Trp Met Trp Lys Val Pro Asn Gly Asp Cys Lys Thr Phe Gin Gly Pro 200 205 210 AAC TGC CCA GCC ACC TGT GGC CGA GTC CTC CCT GAT GGG AAG AGA GCT Asn Cys Pro Ala Thr Cys Gly Arg Val Leu Pro Asp Gly Lys Arg Ala 215 220 225 230 GTG GTA GGC TAT GAA GAT GGG ACC ATC AGG ATT TGG GAC CTG AAG CAG Val Val Gly Tyr Glu Asp Gly Thr lie Arg lie Trp Asp Leu Lys Gin 235 240 245 GGA AGC CCT ATC CAT GTA CTG . AA GGG ACT GAG GGT CAC CAG GGC CCA Gly Ser Pro He His Val Leu -.ys Gly Thr Glu Gly His Gin Gly Pro 250 255 260 CTC ACC TGT GTT GCT GCC AAC CAG GAT GGC AGC TTG ATC CTA ACT GGC Leu Thr Cys Val Ala Ala Asn Gin Asp Gly Ser Leu He Leu Thr Gly 265 270 275 TCT GTG GAC TGC CAG GCC AAG CTG GTC AGT GCC ACC ACC GGC AAG GTG Ser Val Asp cys Gin Ala Lys Leu Val Ser Ala Thr Thr Gly Lys Val 280 285 290 GTG GGT GTT TTT AGA CCT GAG ACT GTG GCC TCC CAG CCC AGC CTG GGA Val Gly Val Phe Arg Pro Glu Thr Val Ala Ser Gin Pro Ser Leu Gly 295 300 305 310 GAA GGG GAG GAG AGT GAG TCC AAC TCG GTG GAG TCC TTG GGC TTC TGC 1 Glu Gly Glu Glu Ser Glu Ser Asn Ser Val Glu Ser Leu Gly Phe Cys 315 320 325 AGT GTG ATG CCC CTG GCA GCT GTT GGC TAC CTG GAT GGG ACC TTG GCC 1 Ser Val Met Pro Leu Ala Ala Val Gly Tyr Leu Asp Gly Thr Leu Ala 330 335 340 ATC TAT ACC TGG CTA CGC AGA CTC TTA GGC ATC AGT GTC AGC ACC AGT 1 He Tyr Thr Trp Leu Arg Arg Leu Leu Gly He Ser Val Ser Thr Ser 345 350 355 CGG GCA TCG TGC AGC TGC TGT GGG AGG CAG GCA CTG CCG TGG TAT ATA Arg Ala Ser Cys Ser Cys Cys Gly Arg Gin Ala Leu Pro Trp Tyr He 360 365 370 CCT GCA GCC TGG ATG GCA TCG TGC GCC TCT GGG ACG CCC GGA CCG GCC Pro Ala Ala Trp Met Ala Ser Cys Ala Ser Gly Thr Pro Gly Pro Ala 375 380 385 390 GCC TGC TTA CTG ACT ACC GGG GCC ACA CGG CTG AGA TCC TGG ACT TTG Ala Cys Leu Leu Thr Thr Gly Ala Thr Arg Leu Arg Ser Trp Thr Leu 395 400 405 CCC TCA GCA AAG ATG CCT CCC TGG TGG TGACCACGTC AGGAGACCAC Pro Ser Ala Lys Met Pro Pro Trp Trp 410 415 AAAGCGAAAG TATTTTGTGT CCAAAGGCCT GACCGTTAAT GGCTGCAGCC CCTGCCTGTG TGTCTGGTGT TGAGGGGACG AAGGGACCCC TGCCCCTGTC TGCCAGCAGA GGCAGTAGGG CACAGAGGGA AGAGGAGGGT GGGGCCCTGG ATGACTTTCC AGCCTCTTCA ACTGACTTGC TCCCCTCTCC TTTTCTTCTC TTTAGAGACC CAGCCCAGGG CCCTCCCACC CTTGCCCAGA CCTGGTGGGC CCTTCAGAGG GAGGGGTGGA CCTGTTTCTC TTTCACTTTC ATTTGCTGGT GTGAGCCATG GGGTGTGTAT TTGTATGTGG GGAGTAGGTG TTTGAGGTTC CCGTTCTTTC CCTTCCCAAG TCTCTGGGGG TGGAAAGGAG GAAGAGATAC TAGTTAAAGA TTTTAAAAAT GTAAATAAAA TATACTTCCC AGAAAAAAAA AAAAAAAAAA AAAAAAA (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 415 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Glu Ser Glu Ser Glu Ser Gly Ala Ala Ala Asp Thr Pro Pro Leu 1 5 10 15 Glu Thr Leu Ser Phe His Gly Asp Glu Glu He He Glu Val Val Glu 20 25 30 Leu Asp Pro Gly Pro Pro Asp Pro Asp Asp Leu Ala Gin Glu Met Glu 35 40 45 Asp Val Asp Phe Glu Glu Glu Glu Glu Glu Glu Gly Asn Glu Glu Gly 50 55 60 Trp Val Leu Glu Pro Gin Glu Gly Val Val Gly Ser Met Glu Gly Pro 65 70 75 80 Asp Asp Ser Glu Val Thr Phe Ala Leu His Ser Ala Ser Val Phe Cys 85 90 95 Val Ser Leu Asp Pro Lys Thr Asn Thr Leu Ala Val Thr Gly Gly Glu 100 105 110 Asp Asp Lys Ala Phe Val Trp Arg Leu Ser Asp Gly Glu Leu Leu Phe 115 120 125 Glu Cys Ala Gly His Lys Asp Ser Val Thr Cys Ala Gly Phe Ser His 130 135 140 Asp Ser Thr Leu Val Ala Thr Gly Asp Met Ser Gly Leu Leu Lys Val 145 150 155 160 Trp Gin Val Asp Thr Lys Glu Glu Val Trp Ser Phe Glu Ala Gly Asp 165 170 175 Leu Glu Trp Met Glu Trp His Pro Arg Ala Pro Val Leu Leu Ala Gly 180 185 190 Thr Ala Asp Gly Asn Thr Trp Met Trp Lys Val Pro Asn Gly Asp Cys 195 200 205 Lys Thr Phe Gin Gly Pro Asn Cys Pro Ala Thr Cys Gly Arg Val Leu 210 215 220 Pro Asp Gly Lys Arg Ala Val Val Gly Tyr Glu Asp Gly Thr He Arg 225 230 235 240 He Trp Asp Leu Lys Gin Gly Ser Pro He His Val Leu Lys Gly Thr 245 250 255 Glu Gly His Gin Gly Pro Leu Thr Cys Val Ala Ala Asn Gin Asp Gly 260 265 270 Ser Leu He Leu Thr Gly Ser Val Asp Cys Gin Ala Lys Leu Val Ser 275 280 285 Ala Thr Thr Gly Lys Val Val Gly Val Phe Arg Pro Glu Thr Val Ala 290 295 300 Ser Gin Pro Ser Leu Gly Glu Gly Glu Glu Ser Glu Ser Asn Ser Val 305 310 315 320 Glu Ser Leu Gly Phe cys Ser Val Met Pro Leu Ala Ala Val Gly Tyr 325 330 335 Leu Asp Gly Thr Leu Ala He Tyr Thr Trp Leu Arg Arg Leu Leu Gly 340 345 350 He Ser Val Ser Thr Ser Arg Ala Ser Cys Ser Cys Cys Gly Arg Gin 355 360 365 Ala Leu Pro Trp Tyr He Pro Ala Ala Trp Met Ala Ser Cys Ala Ser 370 375 380 Gly Thr Pro Gly Pro Ala Ala Cys Leu Leu Thr Thr Gly Ala Thr Arg 385 390 395 400 Leu Arg Ser Trp Thr Leu Pro Ser Ala Lys Met Pro Pro Trp Trp 405 410 415

Claims: WHAT IS CLAIMED IS: 1. A DNA segment coding for a polypeptide comprising an amino acid sequence corresponding to AAMP-l, or at least 5 contiguous amino acids thereof. 2. The DNA segment according to claim 1, wherein said DNA segment comprises the sequence shown in SEQ ID NO:l, allelic or species variation thereof, or at least 15 contiguous nucleotides thereof. 3. The DNA segment according to claim 2, wherein said DNA segment has the sequence shown in SEQ ID NO:l, allelic or species variation thereof. 4. The DNA segment according to claim 3, wherein said DNA segment has the sequence shown in SEQ ID NO:l. 5. The DNA segment according to claim 1, wherein said DNA segment encodes the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof. 6. The DNA segment according to claim 5, wherein said DNA segment encodes the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof. 7. The DNA segment according to claim 6, wherein said DNA segment encodes the amino acid sequence set forth in SEQ ID NO:2. 8. A polypeptide free of proteins with which it is naturally associated and comprising an amino acid sequence corresponding to AAMP-l, or at least 5 contiguous amino acids thereof. 9. The polypeptide according to claim 8, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof. 10. The polypeptide according to claim 9, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof. 11. The polypeptide according to claim 10, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2. 12. A polypeptide bound to a solid support and comprising an amino acid sequence corresponding to AAMP-l. 13. The polypeptide according to claim 12, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof, or at least 5 contiguous amino acids thereof. 14. The polypeptide according to claim 13, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2, allelic or species variation thereof. 15. The polypeptide according to claim 14, wherein said polypeptide comprises the amino acid sequence set forth in SEQ ID NO:2. 16. A recombinant DNA molecule comprising a vector and the DNA segment according to claim 1. 17. A cell that contains the recombinant DNA molecule according to claim 16. IS -±9-. A method of producing a polypeptide having an amino acid sequence corresponding to AAMP-l comprising culturing the cell according to claim 17 under conditions such that said DNA segment is expressed and said polypeptide thereby produced and isolating said polypeptide. IX 20. A vaccine comprising the polypeptide according to claim 8 in an amount effective to elicit protective antibodies in a patient to HIV and a pharmaceutically acceptable diluent, carrier, or excipient. ---- 21. A method of preventing AIDS in a patient comprising administering to said patient the polypeptide according to claim 8 under conditions such that HIV infection is prevented. 22. A therapeutic modality useful in the treatment of inflammatory, immune, or neoplastic disorders in a patient comprising administering to said patient an effective amount of the polypeptide according to claim 8.

Inventors: BECKNER MARIE E, LIOTTA LANCE A, BECKNER, MARIE, E., LIOTTA, LANCE, A.
Applicants: US GOVERNMENT, THE UNITED STATES OF AMERICA, REPRESENTED BY THE S
Application year: 1993
Main classification: A61K37
IPC codes: A61K38, A61P29, A61P35, A61P37, C07K14, C07K16, C12N1, C12N15, C12P21, C12R1, G01N33