MXPA01007077A

MXPA01007077A - Interleukin-17 related mammalian cytokines. polynucleotides encoding them. uses

Info

Publication number: MXPA01007077A
Application number: MXPA/A/2001/007077A
Authority: MX
Inventors: J Fernando Bazan; Robert A Kastelein; Daniel M Gorman
Original assignee: Schering Corporation
Priority date: 1999-01-11
Filing date: 2001-07-11
Publication date: 2002-03-26

Abstract

CTLA-8 related antigens from mammals, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding said antigens. Methods of using said reagents and diagnostic kits are also provided.

Description

MAMMALS CYCOTHES RELATED WITH INTERLEUCINE-17, POLYUCLEOTIDES THAT CODE THEM AND USES FIELD OF THE INVENTION The present invention relates to compositions related to proteins that function in the control of the physiology, development and differentiation of mammalian cells, for example, cells of a mammalian immune system. In particular, it provides nucleic acids, proteins, antibodies and mimetics that regulate the physiology, development and cell differentiation, or the function of several cell types, including hematopoietic cells.

BACKGROUND OF THE INVENTION The immune system of vertebrates consists of a number of organs and several different cell types. Two major cell types include the myeloid and lymphoid lines. Among the lymphoid line are B cells, which were originally characterized as differentiating in fetal liver or adult bone marrow, and T cells, which were originally characterized as differentiating in the thymus. See, for example, Paul (ed. 1998) Fundamental Immunoloqy (4th Ed.) Raven Press, New York.

In many aspects of the development of an immune response or cellular differentiation, the soluble proteins known as cytokines cunplen a decisive function in the regulation of cellular interactions. These cytokines apparently mediate cellular activities in many ways. They have shown, in many cases, to modulate the proliferation, growth and differentiation of hematopoietic stem cells in the vast number of progenitors that make up the lines responsible for an immune response. However, cell molecules that are expressed by different stages of cell development in these maturation pathways are not yet fully identified. In addition, the functions and mechanisms of action of the signaling molecules that induce, sustain or modulate the various physiological, developmental or proliferative states of these cells are poorly understood. Clearly, the immune system and its response to various efforts were important in medicine, for example, in infectious diseases, cancer-related responses and treatment, transplant rejection and allergic responses. See for example, Thorn et al, Harrison's Principles of Internal Medicine MacGraw / Hill, New York. Medical science has, to a large degree, the appropriate recruitment or suppression of the immune system in the effect of cures for inadequate or inappropriate responses to environmental factors. However, the lack of understanding of how the immune system is regulated or differentiated has blocked the ability to conveniently modulate normal defense mechanisms to biological stimulation. Medical conditions characterized by abnormal or inappropriate regulation of the development or physiology of relevant cells consequently can not yet be managed. The discovery and characterization of specific cytokines will contribute to the development of therapies for a wide range of degenerative conditions, or others, that affect the immune system, hematopoietic cells, as well as other cell types. The present invention provides solutions to some of these and many other problems.

BRIEF DESCRIPTION OF THE INVENTION The present invention is based, in part, on the discovery of cDNA clones that encode several cytokine type proteins that exhibit significant sequence similarity to the cytokine designated CTLA-8. The invention encompasses isolated genes encoding the proteins of the invention, variants of the encoded proteins, for example, mutations (muteins) of the natural sequences, allelic and species variants, fusion proteins, chemical mimetics, antibodies and other functional analogues or structural. In addition, various uses of these different protein or nucleic acid compositions are provided.

In certain embodiments of nucleic acid, the invention provides a recombinant or isolated polynucleotide comprising a sequence of: a) a mammalian IL-173, which: encodes at least 8 contiguous amino acids of SEQ ID NO: 6, 8, 10, or 12; encodes at least two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 6, 8, 10 or 12; or comprises one or more segments of at least 21 contiguous nucleotides of SEQ ID NO: 5, 7, 9, or 11; b) a mammalian IL-174, which: encodes at least 8 contiguous amino acids of SEQ ID NO: 14, 16, or 18 encodes at least two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 14, 16 , or 18; or comprises one or more segments of at least 21 contiguous nucleotides of SEQ ID NO: 14, 16, or 18 c) a mammalian IL-176, which: encodes at least 8 contiguous amino acids of SEQ ID NO: 28, codes for at least two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 28; or comprises one or more segments of at least 21 contiguous nucleotides of SEQ ID NO: 27; d) a mammalian IL-177, which: encodes at least 8 contiguous amino acids of SEQ ID NO: 30; encodes at least two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 30; or comprises one or more segments of at least 21 contiguous nucleotides of SEQ ID NO: 29. Other embodiments include said polynucleotide in an expression vector, comprising sequence: a) (IL-173) which: encodes at least 123 amino acids contiguous of SEQ ID NO: 6, 8, 10 or 12; encodes at least two distinct segments of at least 7 and 10 contiguous amino acids of SEQ ID NO: 6, 8, 10, or 12; or comprises at least 27 contiguous nucleotides of SEQ ID NO: 5, 7, 9, 11; b) (IL-174) which: encodes at least 12 contiguous amino acids of SEQ ID NO: 14, 16, or 18; encodes at least two distinct segments of at least 7 and 10 contiguous amino acids of SEQ ID NO: 14, 16, or 18; or comprises at least 27 contiguous nucleotides of SEQ ID NO: 13, 15, or 17; c) (IL-176) which: encodes at least 12 contiguous amino acids of SEQ ID NO: 28; encodes at least two distinct segments of at least 7 and 10 contiguous amino acids of SEQ ID NO: 28; or comprises at least 27 contiguous nucleotides of SEQ ID NO: 27; or d) (IL-177) which: encodes at least 12 contiguous amino acids of SEQ ID NO: 30; encodes at least two distinct segments of at least 7 and 10 contiguous amino acids of SEQ ID NO: 30; or comprises at least 27 contiguous nucleotides of SEQ ID NO: 29. Certain embodiments will include those polynucleotides: a) (IL-173) that: encode at least 16 contiguous amino acid residues of SEQ ID NO: 6, 8, 10, or 12; encode at least two distinct segments of at least 10 and 13 contiguous amino acid residues of SEQ ID NO: 6, 8, 10 or 12; comprise at least 33 contiguous nucleotides of SEQ ID NO: 5, 7, 9, or 11; or comprise the entire mature coding portion of SEQ ID NO: 5, 7, 9, or 11; b) (IL-174) which: encode at least 16 contiguous amino acid residues of SEQ ID NO: 14, 16, or 18; encode at least two distinct segments of at least 10 and 13 contiguous amino acid residues of SEQ ID NO: 14, 16 or 18; comprise at least 33 contiguous nucleotides of SEQ ID NO: 13, 15, or 17; or comprise the entire mature coding portion of SEQ ID NO: 13, 15, or 17; c) (IL-176) which: encodes at least 16 contiguous amino acids of SEQ ID NO: 28; encode at least two distinct segments of at least 10 and 14 contiguous amino acid residues of SEQ ID NO: 28; comprise at least 33 contiguous nucleotides of SEQ ID NO: 27; or comprise the entire mature coding portion of SEQ ID NO: 27; or d) (IL-177) which: encode at least 16 contiguous amino acids of SEQ ID NO: 30; encode at least two distinct segments of at least 10 and 14 contiguous amino acid residues of SEQ ID NO: 30; comprises at least 33 contiguous nucleotides of SEQ ID NO: 29; or comprise the entire mature coding portion of SEQ ID NO: 29. Various methods are provided, for example, to make: a) a polypeptide comprising expressing the described expression vector, thereby producing the polypeptide; b) a double nucleic acid comprising contacting a described polynucleotide with a complementary nucleic acid, thereby resulting in the production of the double nucleic acid; or c) a described polynucleotide that comprises amplifying using a PCR method. Alternatively, the invention provides a recombinant or isolated polynucleotide that hybridizes under severe wash conditions of at least 55 ° C and less than 400 mM salt, to: a) the described polynucleotide (IL-173) consisting of the coding portion of SEQ ID NO: 5, 7, 9, or 11; b) the described polynucleotide (IL-174) consisting of the coding portion of SEQ ID NO: 13, 15, or 17; the described polynucleotide (IL-176) consisting of the portion of SEQ ID NO: 27; or d) the described polynucleotide (IL-177) consisting of the coding portion of SEQ ID NO: 29. Other embodiments include said described polynucleotide; a) where the washing conditions are at least 65 ° C and less than 300 mM salt; or b) comprising at least 50 contiguous nucleotides of the coding portion of SEQ ID NO: 5, 7, 9, or 11 (IL-173); SEQ ID NO: 13, 15, or 17 (IL-174); SEQ ID NO: 27 (IL-176), or SEQ ID NO: 29 (IL-177). Certain kits are provided, for example, comprising a described polynucleotide, and: a) instructions for the use of the polynucleotide for detection; b) instructions for the disposal of the polynucleotide or other reagents of the equipment; or c) both a and b. In addition, several cells are provided, for example, a cell containing the described expression vector, wherein the cell is: a prokaryotic cell; a eukaryotic cell; a bacterial cell; a yeast cell; an insect cell; a mammalian cell; a mouse cell; a primate cell; or a human cell. Modes of polypeptide include, for example, an isolated or recombinant antigenic polypeptide: a) (IL-173) comprising at least: i) a segment of 8 identical contiguous amino acids of SEQ ID NO: 6, 8, 10, or 12; or ii) two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 6, 8, 10, or 12; c) (IL-174) comprising at least: i) a segment of 8 identical contiguous amino acids of SEQ ID NO: 14, 16, or 18; or ii) two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 14, 16, or 18; c) (IL-176) comprising at least: i) a segment of 8 identical contiguous amino acids of SEQ ID NO: 28; or ii) two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 28; or d) (IL-177) comprising at least: i) a segment of 8 identical contiguous amino acids of SEQ ID NO: 30; or ii) two distinct segments of at least 5 contiguous amino acids of SEQ ID NO: 30. Additional moieties include said polypeptide described, wherein a) the segment of 8 identical contiguous amino acids is at least 14 contiguous amino acids; or b) one of the segments of at least 5 contiguous amino acids comprises at least 7 contiguous amino acids. Other embodiments include a described polypeptide, wherein. A) (IL-173) the polypeptide: a) comprises a mature sequence of SEQ ID NO: 6, 8, 10, or 12; b) binds selectively to a polyclonal antibody raised against an immunogen of a mature SEQ ID NO: 6, 8, 10, or 12; c) comprises a plurality of polypeptide segments other than 10 contiguous amino acids of SEQ ID NO: 6, 8, 10 or 12; d) is a natural allelic variant of SEQ ID NO: 6, 8, 10 or 12; e) has a length of at least 30 amino acids; or f) exhibits at least two non-overlapping epitopes, which are selective for mature SE ID: NO: 6, 8, 10 or 12; B) (IL-174) the polypeptide: a) comprises mature SEQ ID NO: 14, 16 or 18; b) binds selectively to a polyclonal antibody generated against an immunogen of mature SE ID NO. 14, 16, or 18; c) comprises a plurality of polypeptide segments other than 10 contiguous amino acids of SEQ ID NO: 14, 16 or 18; d) has a length of at least 30 amino acids; or e) exhibits at least two non-overlapping epitopes, which are selective for mature SE ID: 14, 16 or 18; or D) (IL-176) the polypeptide: a) comprises SEQ ID NO: 28; b) binds selectively to a polyclonal antibody generated against an immunogen of SEQ ID NO: 28; c) comprises a plurality of polypeptide segments other than 10 contiguous amino acids of SEQ ID NO: 28; d) has a length of at least 30 amino acids; or e) exhibits at least two non-overlapping epitopes, which are selective for the primate protein of SEQ ID NO: 28; or D) (IL-177) the polypeptide: a) comprises SEQ ID NO: 30; b) binds selectively to a polyclonal antibody generated against an immunogen of SEQ ID NO: 30; c) comprises a plurality of polypeptide segments other than 10 contiguous amino acids of SEQ ID NO: 30; d) has a length of at least 30 amino acids; or e) exhibits at least two non-overlapping epitopes, which are selective for the primate protein of SEQ ID NO: 30. Various other embodiments include said described polypeptide, which: a) is in a sterile composition; b) it is not glycosylated; c) is denatured; d) is a synthetic polypeptide; e) is attached to a solid substrate; f) is a fusion protein with a detection or purification tag; g) is a substitution of 5 folds or less of a natural sequence; or h) is a deletion variant or insertion of a natural sequence. Further provided are methods for the use of the polypeptides that are described, for example, a) for labeling the polypeptide, which comprises labeling the polypeptide with a radioactive label; b) to separate the polypeptide from another polypeptide in a mixture, which comprises running the mixture in a chromatography matrix, thus separating the polypeptides; c) to identify a compound that selectively binds to the polypeptide, which comprises incubating the compound with the polypeptide under appropriate conditions; thereby causing the compound to bind to the polypeptide; or d) to conjugate the polypeptide to a matrix, which comprises derivatizing the polypeptide with a reactant that reacts, and conjugating the polypeptide to the matrix. In addition antibodies are provided, including a binding compound comprising an antigen binding portion of an antibody that binds selectivity to said described polypeptide, wherein the polypeptide: a) (IL-173) comprises the mature polypeptide of SEQ ID NO: 1. NO: 6, 8, 10 or 12; b) (IL-174) comprises SEQ ID NO: 14, 16 or 18; c) (IL-176) comprises SEQ ID NO: 28; od) (IL-177) comprises SEQ ID NO: 30. Certain embodiments encompass said binding compound, wherein the antibody is a polyclonal antibody that is raised against the polypeptide of: a) (IL-173) SEQ ID NO: 6 , 8, 10 or 12; b) (IL-174) SEQ ID NO: 14, 16, or 18; c) (IL-176) SEQ ID NO: 28; or d) (IL-177) SEQ ID NO: 30. Other embodiments include said described linking compound, wherein: a) antibody: i) is immunoselected; ii) binds to a denatured protein; or iii) exhibits a Kd to the polypeptide of at least 30 mM; or b) the linking compound: i) binds to a solid substrate, including a bead or plastic membrane; ii) is in a sterile composition; or ii) is detectably labeled, including a radioactive or fluorescent label. Methods are provided, for example by producing an antigen: antibody complex, comprising contacting a polypeptide comprising a sequence of SEQ ID NO: 6, 8, 10, 12, 14, 16, 18, 28 or 30, with a compound bond described, under conditions that allow the complex to be formed. Preferably, the binding compound is an antibody, and the polypeptide is in a biological sample. Equipment is provided, for example, comprising a described link compound and: a) a polypeptide of SEQ ID NO: 6, 8, 10, 12, 14, 16, 18, 28 or 30; b) instructions for the use of the binding compound for detection; or c) instructions for the disposal of the binding compound or other equipment reagents. And a method is provided for evaluating the binding selectivity of an antibody to a protein of SEQ ID NO: 6, 8, 10, 12, 14, 16, 18, 28 or 30, which comprises contacting a described antibody with the protein, and with another cytokine; and compare the binding of the antibody to the protein and to the cytokine.

DETAILED DESCRIPTION OF THE PREFERRED MODALITIES 1. General The present invention provides DNA sequence encoding several mammalian proteins that exhibit structural characteristics characteristic of cytokines, in particular related to the cytokine designated CTLA-8 (also referred to as IL-17). The human, rat, mouse and a viral homolog form of CTLA-8 have been described, and their sequences are available from GenBank. See Rouvier et al. (1993) J. Immunol. 150: 5445-5456; Yao et al. (1995) Immunity 3: 811-821 Yao et al. (1995) J. Immunol. 155: 5483-5486; and Kennedy et al. (1996) J. Interferon and Cytoquine Res. 16: 611-617. CTLA-8 has activities involved in arthritis, kidney graft rejection, tumorigenicity, host-virus interactions and innate immunity; and it seems to exhibit certain regulatory functions similar to IL-6. See PubMed (search for IL-17); Chabaud et al. (1998) J. Immunol. 63: 139-148; Amin et al. (1998) Curr. Opin. Rheumatol. 10: 263-268; Van Kooten et al. (1998) J. Am. Soc. Nephrol 9: 1526-1534; Fossiez et al. (1998) Int. Rev. Immunol. 16: 541-551; Knappe et al. (1998) J. Virol. 72: 5797-5801; Seow (1998) Vet. Immuno Immunopathol. 63: 139-48; and Teunissen et al. (1998) J. Invest. Dermatol. 111: 645-649. A report on signaling through the transcription factor NFkB involves a signal path that uses innate immunity. Shalom-Barak et al. (1998) J. Biol. Chem. 273: 27467-27473.

The recently presented cDNA sequences exhibit several characteristics that are characteristic of mRNA encoding cytokines, growth factors and oncogenes. Because IL-17 is the first member of this newly recognized family of cytokines related to TGF-β, the applicants have designated the IL-170 family, with the new members IL-172, IL-173, IL-174, IL-176, IL-177; and IL-171 and IL-175. It is predicted that the fold for this family is that of the TGF-β family of cytokines. The TGF-β family of cytokines, and the IL-170 family share the common feature of a cysteine knot motif, characterized by a particular spacing of cysteine residues. See, for example, Sun and Davies (1995) Ann. Rev. Biophvs. Biomolec. Struct. 24: 269-291; McDonald et al. (1993) Cell 73; 421-424; and Isaacs (1995) Curr. Op. Struct. Biol. 5: 391-395. In particular, the structures suggest a number of conserved cysteines, which correspond, and are numbered, in human IL-172 (SEQ ID NO: 2), to the cysteines in 101, 103, 143, 156 and 158. The first cysteine corresponds to the position in table 7 of human IL-172 (SEQ ID NO: 2) va119. The fourth cysteine corresponds to that of mouse IL-172 (SEQ ID NO: 4) cys141; in human IL-173 (SEQ ID NO: 6) cys119; in mouse IL-174 (SEQ ID NO: 16) cys104; and in human IL-171 (SEQ ID NO: 21) cys50. The disulfide bonds should be cysteines 2 with 5; and 3 with 6; and 1 with 4. The functional significance of fold similarity suggests the formation of dimers for the IL-70 family. As a consequence, IL-70 dimers would couple two cell surface receptors, through which signal transduction will occur.

These new proteins are designated related to CTLA-8, or generally IL-70 proteins. Natural proteins should be able to mediate several physiological responses that would lead to biological or physiological responses in target cells, for example, those responses characteristics of cytokine signaling. Initial studies have located the message that encodes this protein in several cell lines of hematopoietic cells. The genes encoding the original CTLA-8 antigen (IL-17) have been mapped to mouse chromosome 1A and human chromosome 2q31. Murine CTLA-8 was originally cloned by Rouvier et al. (1993) J. Immunol. 150: 5445-5456. Human IL-173 has been mapped on chromosome 13Q11. Similar sequences for proteins in other mammalian species should also be available. Purified CTLA-8, when grown with synoviocytes, it is able to induce the secretion of IL-6 from these cells. This induction is reversed with the addition of a neutralizing antibody produced against human CTLA-8. Endothelial, epithelial, fibroblast and carcinoma cells also exhibit responses to treatment with CTLA-8. This information suggests that CTLA-8 may be involved in inflammatory fibrosis, for example, psoriasis, scleroderma, pulmonary fibrosis, or cirrhosis. CTLA-8 can also cause the proliferation of carcinomas or other cancer cells, since IL-6 often acts as a growth factor for these cells. As such, the other members of the recently discovered related family probably have similar or related biological activities.

The descriptions below are directed, for exemplary purposes, to a human or murine IL-170 protein, but are equally applicable to related modalities of other species.

II: Nucleic acids Tables 1-6 describe the nucleotide and amino acid sequences of several new sequences of members of the IL-170 family. The nucleotide sequences that are described and the related reagents are useful in the construction of DNA clones useful for extending the clones in both directions for flanking or full-length sequence determination, expressing IL-170 polypeptides, or for example, isolating a homologous gene from another natural source. Typically, the sequences will be useful for isolating other genes, e.g., allelic, mouse variants, and similar procedures will be applied to isolate genes from other species, e.g., warm-blooded animals, such as birds and mammals. Cross-hybridization will allow the isolation of genes from other species. A number of different approaches should be available to successfully isolate a suitable nucleic acid clone from another source. Table 1: nucleotide sequence encoding a primate IL-172 polypeptide, eg, human, and predicted amino acid sequence.

In addition, you can use complementary nucleic acid sequences for many purposes. Predicted signal decomposition site indicated, but there may be some debris on either side; putative glycosylation site at residues 55-57. SEQ ID NO: 1 and 2.

° ATG GAC TGG CCT CAC AAC CTG CTG TTT CTT CTT ACC ATT TCC ATC TTC 43 Met Asp Trp Pro His Asn Leu Leu Phe Leu Leu Thr lie Ser lie Phe • -20 -15 -10 -5 CTG GGG CTG GGC CAG CCC AGG AGC CCC AAA AGC A? G AGG AAG GGG CAA 95 Leu Gly Leu Gly Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln 1 5 10 GGG CGG CCT GGG CCC CTG GTC CCT GGC CCT CAC CAG GTG CCA CTG GAC] _44 Gly Arg Pro Gly Pro Leu Val Pro Gly Pro His Gln Val Pro Leu Asp 15 20 25 CTG GTG TCA CGG ATG AAA CCG TAT GCC CGC ATG GAG GAG TAT GAG AGG 192 -Í G \ Leu Val Ser Arg Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg 30 35 40 AAC ATC GAG GAG ATG GTG GCC CAG CTG AGG AAC AGC TCA GAG CTG GCC 240 Asn lie Glu Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing 45 50 55 60 CAG AGA AAG TGT GAG GTC AAC TTG CAG CTG TGG ATG TCC AAC AAG AGG 288 Gln Arg Lys Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg 65 70 75 AGC CTG TCT CCC TGG GGC TAC AGC ATC? AC CAC GAC CCC AGC CGT ATC 335 Ser Leu Ser Pro Trp Gly Tyr Ser lie Asn Hxs Asp Pro Ser Arg lie 80 85 90 CCC GTG GAC CTG CCG GAG GCA CGG TGC CTG TGT CTG GGC TGT GTG AAC 334 Pro Val Asp Leu Pro Glu Ala Arg Cys Leu Cys Leu Gly Cys Val Asn 95 100 105 CCC TTC ACC ATG CAG GAG GAC CGC AGC ATG GTG AGC GTG CCG GTG TTC 4 2 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Va] Ser Val Pro Val Phe 110 115 120 AGC CAG GTT CCT GTG CGC CGC CGC CTC TGC CCG CCA CCG CCC CGC ACA 480 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr 125 130 135 140 0 GGG CCT TGC CGC CAG CGC GCA GTC ATG GAG ACC ATC GCT GTG GGC TGC 528 Gly Pro Cys Arg Gln Arg Wing Val Met Glu Thr He Wing Val Gly Cys 145 150 155 ACC TGC ATC TTC TCA 543 Thr Cys He Phe. "160 MDWPHNLLFLLTISIFLGLG QPRSPKSKRKGQGRPGPLVPGPHQVPLDLVSRMKPY? RMEEYER IEEMVAQLRNSSELAQRKCEV LQLWMSIJKRSLSPWGYSTNHDPSRIPVDLPEARCLCLGCV PFT MQEDRSMVSVPVFSQVPVRRRLCPPPPRTGPCRQRAVMETIAVGCTCIF Particularly interesting segments include, for example, example, those that start or end with g1 n1; va119; pro20; pro22, Iys34; pro35; Ieu78; ser79; glu98; ala99; phe110; thr111; cys143; or arg 144. Nucleotide sequence encoding a rodent IL-172 polypeptide, e.g., mouse, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. Predicted signal decomposition site indicated, but there may be some debris on either side; glycosylation site putative in residues 53-55. SEQ ID NO: 3 and 4.

ATG GAC TGG CCG CAC AGC CTG CTC TTC CTC CTG GCC ATC TCC ATC TTC 48 Met Asp Trp Pro His Ser Leu Leu Phe Leu Leu Ala He Ser He Phe -22 -20 -15 -10 CTG GCG CCA AGC CAC CCC CGG AAC ACC AAA GGC AiA AGA AAA GGG CAA 96 Leu Ala Pro Ser His Pro Arg Asn Thr Lys Gly Lys Arg Lvs Gly Gln -5 1 5"10 GGG AGG CCC AGT CCC TTG GCC CCT GGG CCT CAG GTG CCG CTG GAC 144 Gly Arg Pro Ser Pro Leu Ala Pro Giv Pro His Cln Val Pro Leu Asp 15 20 25 CTG GTG TCT CGA GTA AAG CCC TAC GCT CGA ATG GAA GAG TAT GAG CGG 192 Leu Val Ser Arg Val Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg -30 '35 40 AAC CTT GGG GAG ATG GTG GCC CAG CTG AGG AAC AGC TCC GAG CCA GCC! 40 Asn Leu Gly Glu Met Val Ala Gln Leu Arg Asn Ser Ser Glu Pro Wing 45 50 55 AAG AAG AAA TGT GAA GTC AAT CTA CAG CTG TGG TTG TCC AAC AAG AGG 288 Lys Lys Lys Cys Glu Val Asn Leu Gn Leu Tro Leu Ser Asn Lvs Ars 60 65 '70"AGC CTG TCC CCA TGG GGC TAC AGC ATC AAC CAC GAC CCC AGC CGC ATC 336 Ser Leu Ser Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He 75 80 85 90 CCT GCG GAC TTG CCC GAG GCG CGG TGC CTA TGT TTG GGT TGC GTG AAT 384 Pro Wing Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Glv Cvs Val Asn 95 100"" 105 CCC TTC ACC ATG CAG GAG GAC CGT AGC ATG GTG AGC GTG CCA GTG TTC 432 Pro Phe Thr Met Gln Glu Aso Arg Ser Mee Val Ser Val Pro Val Phe 130 115 120 AGC CAG GTG CCG GTG CGC CGC CGC CTC TGT CCT CA. CCT CCT CGC CCT 480 Ser Gln Val Pro Val Arg Arg? Rg Leu Cys Pro Gln Pro Pro Ara Pro 125 130 135 GGG CCC TGC CGC CAG CGT GTC GTC ATG GAG ACC ATC G ^ T GTG GGT TGC 528 Gly Pro Cys Arg Gln Arg Val Val Met Glu Thr Ue Wing Val Glv Cvs 140 145 iso ACC TGC ATC TTC TGA 5 3 Thr Cys He Phe 155 M WPHSLLFLLAI SIFLAPSHP FJKITKGKRKGQGRPS PLAPGPHQVPLDI? 'SRVKPYARMEEYERN LGEMVAQLRNSSEPAKKKCEV LQLWLSNKRSLSPWGYSINHDPSRIPAOLPEARCLCLGCV? PFT MQEDRSMVSVPVFSQVPVRRRLCFQPPRPGPCRQRVVMETIAVGCTC ^ F Particularly interesting segments include, for example, those that start or end with argl; alai 7; pro18; pro20; his21; Iys32; pro33; Ieu76; ser77; glu96; ala97; phe108; thr109; cys141; or arg 142. Table 2: nucleotide sequence encoding a primate IL-173 polypeptide, eg, human, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 5 and 6.

TGC GCG GAC CGG CCG GAG GAG CTA CTG GAG CAG CTG TAC GGG CGC CTG Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu 1 5 10 15 rrc GCC GGC GTG CTC AGT GCC TTC CAC CAC ACG CTG CAG CTG GGG CCG 96 AS Sía GS Va? Leu Be Wing Phe Kis His Thr Leu Gln Leu Gly Pro 20 25 a? CGT GAG CAG GCG CGC AAC GCG AGC TGC CCG GCA GGG GGC AGG CCC GCC 144 Arg Glu Gln Ala Arg Aen Wing Ser Cys Pro Wing Gly Gly Arg Pro Wing 35 40 45 GAC CGC CGC TTC CGG ACG CCC ACC AAC CTG CGC AGC GTG TCG CCC TGG 192 Asp Arg Arg Phe Arg Thr Pro Thr Asn Leu Arg Ser Val Ser Pro Trp 50 55 60 GCC TAC AGA ATC TCC TAC GAC CCG GCG AGG TAC CCC AGG TAC CTG CCT 240 Wing Tyr Arg He Ser Tyr Asp Pro Wing Arg Tyr Pro Arg Tyr Leu Pro 65 70 75 80 GAA. GCC TAC TGC CTG TGC CGG GGC TGC CTG ACC GGG CTG TTC GGC GAG 288 Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr Giy Leu Phe Glv Glu 85 90 95 GAG GAC GTG CGC TTC CGC AGC GCC CCT GTC TAC ATG CCC ACC GTC GTC 336 Glu Asp Val Arg Phe Arg Ser Wing Pro Val Tyr Mee Pro Thr Val Val 100 105 110 CTG CGC CGC ACC CCC GCC TGC GCC GGC GGC CGT TCC GTC TAC ACC GAG 384 Leu Arg Arg Thr Pro Wing Cys Wing Gly Gly Arg Ser Val Tyr Thr Glu 115 120 125 GCC TAC GTC ACC ATC CCC GTG GGC TGC ACC TGC GTC CCC GAG CCG GAG 432 Wing Tyr Val Thr He Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu 130 135 140 AAG GAC GAC GAC AGC ATC AAC T 454 Asp Lys Asp Ala Ser Asn 145 150 CADRPEELLEQLYGRLAAGVLSAFHHTLQLGPREQARNA I? CPAGGRPADRRFRTPTNLRS VSPW? YRISYDPARYPRYLPEAYCLCRGCLTGLFGEEDVRFRSAPVYMPTWLRRTPACA GGRSV7TEAYVTIPVGCTCVPEPEKDADSIN additional nucleotide sequence encoding an IL-173 primate, e.g., human, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 7 and 8 gcccgggcag gtggcgacct cgctcagtcg gcttctcggt ccaegtcccc gggtctgg 58 atg ctg gta gcc gcc ctg ctg gcg ctg ccg ceg age tgc gcc gcg 106 Met Leu Val Wing Gly -Phe Leu Leu Ala Pro Leu Pro Ser Tr-p Ala Wing -15 -10 -5 ggc gcc ceg ags gcg ggc agg cgc ecc gcg cgg ceg cgg ggc tgc gcg 154 GÍy Ala Pro Arg Ala Gly Arg Arg Pro Ala Arg Pro Arg Gly Cys Ala -1 1 5 10 15 gag cgg gag gag cta cg cg gag cag ctg tac ggg cgc ctg gcg gcc 202 Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu Wing Ala 20 25. 30 ggc gtg etc agt gcc ttc falls falls acg ctg cag ctg ggg ceg cgt gag 250 Gly Val Leu Ser Wing Phe His His Thr Leu Gln Leu Gly Pro Arg Glu 35 40 45 cag gcg cgc aac gcg age tgc ceg gca ggg ggc agg ecc gcc gac cgc 298 Gln Wing Arg Asn Wing Cys Pro Wing Gly Gly Arg Pro Wing Asp Arg 50 55 60 cgc ttc cgg ceg ecc acc aac ctc cgc age gtg teg ecc tgg gcc tac 346 Arg Phe Arg Pro Pro Thr Asn Leu Arg Ser Val Ser Pro Trp Wing Tyr 65 70 75 aga ate tec tac gac ceg icg agg tac ecc agg tac ctg ect gaa gcc 394 Arg He Ser Tyr Asp Pro Ala Arg Tyr Pro Arg Tyr Leu Pro Glu Wing 80 85 90 95 tac tgc ctg tgc cgg ggc tgc ctg acc ggg ctg ttc ggc gag gag gac 442 Tyr Cys Leu Cys Arg Gly Cys Leu Thr Gly Leu Phe Gly Glu Glu Asp 100 105 110 9t9 cgc tcc cgc age gcc ect gtc tac atg ecc acc gtc gtc ctg cgc 490 Val Arg Phe Arg Ser Wing Pro Val Tyr Met Pro Thr Val Val Leu Arg 115 120 125 cgc acc ecc gcc tgc gcc ggc ggc cgt tec gte tac acc gag gcc tac 538 Arg Thr Pro Ala Cys Wing Gly Gly Arg Ser Val Tvr Thr Glu Ala Tyr 130 135 140 gtc acc ate ecc gtg ggc tgc acc tgc gtc ecc gao ceg gag aag gac 586 Val Thr He Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu Lys Asp 145 150 155 gca gac age ate aac tec age ate gac aaa cag ggc gcc aag etc ctg 634 Wing Aep Ser He Asn Ser Be He Asp Lys Gln Gly Wing Lys Leu Leu 160 155 170 175 ctg ggc ecc aac gac gcg ecc gct gcc ecc tgaygccggt cctgccccgg 684 Leu Gly Pro Asn Asp Ala Pro Wing Gly Pro 180 185 gaggtctccc cggcccgcat cccgaggcgc ccaagctgga sccgcctgga gggctcggtc 744 ggcgacctct gaagagagtg caccgagcaa accaagtgcc gcagcaccag cgccgccttt 804 ccatggagac tcgtaagcag cttcatctga cacgggcatc ccíggcttgc ttttagctac 864 aagcaagcag cgtggctgga agctgatggg aaacgacccg gcacgggcat cctgtgtgcg 924 gcccgcatgg agggtttgga aaagttcacg gaggctccct gaggagcctc tcagatcggc 984 tgctgcgggt gcagggcgtg actcaccgct gggtgcttgc: caaagagata gggacgcata 1044 agcaatctaa tgctttttaa aaataataat aagtatagcg to tatatacc tacttttaaa 1104 atcaactgtt ttgaatagag gcagagctat tttatattat caaatgagag ctactctgtt 1164 acatttetta acatataaac a tcgtttttt acttcttctg gtagaatttt ttaaagcata 1224 attggaatcc ttggataaat tttgtagctg gtacactctg gcctgggtct ctgaattcag 1284 cctgtcaccg atggctgact gatgaaatgg acacgtctca tctgacccac tcttccttcc 1344 actgaaggtc ttcacgggcc tccaggcctc gtgccgaatt c 1385 MLVAGFLLALPPSWAAGAPRAGRRPARPRGCADRPEELLEQLYGRLAAGVLSAFHHTLQLGPREQARNA SCPAGGRPADRRFRPPTNLRSVSPWAYRISYDPARYPRYLPEAYCLCRGCLTGLFGEEDVRFRSAPVYM PTWLRRTPACAGGRSV? T? AYVTIPVGCTCVPEPEKDADST SSIDKQGAKLLLGPNDAPAGP Important predicted reasons include, for example, cAMP PK at 50-53, 66-69, 72-75 and 113-116; Ca Phos at 82-84 and 166-168; myristoyzalyl sites at 57-61 and 164-166; phosphorylation sites at 50, 53, 72, 75, 80, 82, 113 and 116.

Nucleotide sequence encoding a rodent IL-173 polypeptide, eg, rat, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 9 and 10.

TTT CCG AGA TAC CTG CCC GAA GCC TAC TGC CTG TGC CGA GGC TGT CTG 48 Phe Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu 1 5 10 15 ACC GGG CTC TAC GGT GAG GAC GAC TTC CGC TTT CGC AGC GCA CCC GTC 96 Thr Gly Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Wing Pro Val 20 25 30 TTC TCT CCG GCG GTG GTG CTG CGG CGC ACG GCG GCC T 133 Phe Ser Pro Wing Val Val Leu Arg Arg Thr Wing Ala 35 40 FPRYLPEAYCLCRGCLTGLYGEEDFRFRSAPVFSPAWLRRTAA Supplemental nucleotide sequence encoding a rodent IL-173 polypeptide, eg, mouse, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 11 and 12 atg ttg ggg here ctg gtc tgg atg ecc etc gtc ggc ttc ctg ctg gca 48 Met Leu Gly Thr Leu Val Trp Met Leu Leu Val ¿? and Phe Leu Leu Ala -20 -15 -10 ctg gcg ceg ggc cgc gcg gcg ggc gcg ctg agg acc ggg agg cgc cgc ceg 96 Leu Wing Pro Gly Arg Wing Wing Gly Wing Leu Arg Thr Gly Arg Arg Pro -5 -1 1 5 gcg cgg ceg cgg gac tgc gcg gac cgg cea gag gag etc ctg gag cag 144 Wing Arg Pro Arg Asp Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln 10 15 20 ctg tac ggg cgg cg gcg gcg gcg czc aq? Cc ttc falls falls acg 192 Leu Tyr Gly Arg Leu Wing Wing Gly Val Leu Ser Wing Phe His His Thr 25 30 35 40 ctg cag etc ggg ceg cgc gag cag gcg cgc aat. gcc age tgc ceg gcc 240 Leu Gln Leu Gly Pro Arg Glu Gin Wing Arg Asn Wing Being Cys Pro Wing 5 50 55 ggg ggc agg gcc gcc gac cgc cgc ttc cgg cea ecc acc aac ctg cgc 288 Gly Gly Arg Wing Wing Asp Arg Arg Phe Arg Pro Pro Thr Asn Leu Arg 60 G5 70 age gtg teg ecc tgg gcg tac agg att tec tac gac ect gct cgc ttt 336 Ser Val Ser Pro Trp Wing Tyr Arg He Ser Tyr Aso Pro Wing Arg Phe 75 80 85 ceg agg tac ctg ecc gaa gcc tac tgc ctg tgc cga ggc tgc ctg acc 384 Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr 90 95 100 ggg etc drops ggg gag gag gac tcc cgc ttt cg ~ age here ecc gtc ttc 432 Ga and Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Thr Pro Val Phe 105 110 115 120 tet cea gcc gtg gtg ctg cgg cgc asa gcg gcc cgc gcg ggc ggc cgc 480 Ser Pro Ala Val Val Leu Arg Ai g Thr Ala A-la Cys Ala Gly Gly Arg 125 130 135 tet gtg tac gcc gaa falls tac ate acc ate ceg gtg ggc tgc acc tgc 528 Ser Val Tyr Ala Clu His Tyr He Thr He Pro Val Gly Cys Thr Cys 140 145 150 gtg ecc gag ceg gac aag tec gcg gac agt gcg aac tec age atg gac 576 Val Pro Glu Pro Asp Lys Ser Wing Asp Ser Wing Asn Ser Ser Met Asp 155 160 165 aag ctg ctg ggg ecc gcc gac agg ect gcg ggg cgc tgatgccggg 625 Lys Leu Leu Gly Pro Wing Asp Arg Pro Al Gly Arg 170 _175 180 gactgcccgc catggcccag cttcctgcat gcatcaggtc ccctggccct gacaaaaccc 685 accccatgat ccctggccgc tgcctaattt ttccaaaagg acagetacat aagctttaaa 745 aaagtagaca tatatttttc etacatatet acaactattt tgaatagtgg cagaaactat 805 gtaatttaga ttteatatta gcaagcatgt tgtttttaaa cttctttgat atacaageac 865 atcacacaca tcccgttttc ctctagtagg attcttgagt gcataattgt agtgetcaga 925 tgaacttcct tctgctgcac tgtgccctgt ccctgagtct ctcctgtggc ccaagcttac 985 taaggtgata atgagtgctc cggatctggg cacctaaggt ctccaggtcc ctggagaggg 1045 agggatgtgg gggggctagg aaccaagcgc ccctttgttc tttagcttat ggatggtctt 1105 aactttataa aga taaagt ttttggtgtt attctttc 1143 MLGTLVV7MLLVGFLLiAPGRAJGALRTGRRPARPRDCADRPEE L.EQLYGRLAAGVLSAFHHTLQLGPRE QARNASCPAGGRAADRRFRPPTNLRSVSPWAYRISYDPARFPRY.PEAYCLCRGCLTGLYGEEDFRFRSTP VFS PAWLRRTAACAGCRSVYAEHYITI PVGCTCVPEPDKSADSANSSMDKLLLGPADRPAGR.

Important predicted reasons include, for example, cAMP PK sites at 50-53, 66-69, 72-75 and 113-116; Ca phosphorylation sites at 82-84, 159-161 and 166-168; myristoyllil sites at 57-61 and 101-105; N-glycosyl sites at 51-53 and 164-166; phosphorylation sites at 50, 53, 72, 75, 80, 82, 113 and 116; and PKC phosphorylation sites at 4-6. Table 3: nucleotide sequence encoding a primate IL-174 polypeptide, eg, human, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 13 and 14. tgagtgtgca gtgccagc atg tac cag gtg gtt gca ttc ttg gca atg gtc 51 Met Tyr Gln Val Val Ala Phe Leu Ala Met Val -15 -10 atg gga acc fall acc tac age falls tgg ecc age cgc tgc ecc age aaa 99 Met Gly Thr His Thr Tyr Ser His Trp Pro Ser Cys Cys Pro Ser Lys -5 -1 1 5 10 ggg cag gac acc tet gag gag ctg ctg agg tgg age act gtg ect gtg 147 Gly Gln Asp Thr Ser Glu Glu Leu Leu Arg Trp Ser Thr Val Pro Val 15 20 25 ect ecc cta gag -ect gct agg ecc aac cgc cae cea gag tec tgt agg 195 Pro Pro Leu Glu Pro Ala ^ rg Pro Asn Arg H? S Pro Glu Ser Cys Arg 30 35 40 gcc agt gaa gat gga ecc etc aac age agg gcc ate tec ecc tgg aga 243 Wing Ser Glu Asp Gly Pro Leu Asn Ser Arg Wing He Ser Pro Trp Arg 45 50 55 tat gag ttg gac aga gae ttg aac cgg etc ecc cag gac ctg tac falls 291 Tyr Glu Leu Asp Arg A.sp Leu Asn Arg Leu Pro Gln A.sp Leu Tyr His 60 65 70 75 gcc cgt tgc ctg tgc ceg falls tgc gtc age cta cag here ggc tec drops 339 Wing Arg Cys Leu Cys Pro His Cys Val Ser Leu Gln Thr Gly Ser His 80 85 90 atg gac ecc cgg ggc aac teg gag ctg etc tac falls aac cag act gtc 387 Met Asp Pro Arg Cly A.sn Ser Glu Leu Leu Tyr His Asn Gln Thr Val 95 100 105 ttc tac cgg cgg cg tgc cat ggc gag aag ggc acc cae aag ggc tac 435 Phe Tyr Arg Arg Pro Cys His Gly Glu Lys Gly Thr His Lys Gly Tyr 110 115 120 tgc ctg gag cgc agg ctg tac cgt gtt tec t ta gct tgt gtg tgt gtg 483 Cys Leu Glu Arg Arg Leu Tyr Arg Val Ser Leu Ala Cys Val Cys Val 125 130 135 cgg ecc cgt gtg atg ggc tag Arg Pro Arg Val Met Gly 140 145 MYQWAFLAMVMGTHTYSHWPSCCPSKGQDTSEELLRWSTVPVPPLEPARPNRHPESCRASEDGPL NSRAI S P RYELDRDL RLPQDLYHARCLCPHCVSLQTGSHMDPRGNSELLYHNQTVFYRRPCHGE KGTHKGYCLERRLYRVSLACVCVRPRVMG Important predicted reasons include, for example, cAMP PK sites at 21-24, 53-56 and 95-98; Ca phosphorylation sites at 15-17, 16-18 and 45-47; myristoyzalyl sites at 12-16, 115-119 and 118-122; N-glycosyl site at 104-107; phosphorylation sites in 21, 23, 43, 53, 56, 95, 98 and 131; PKC phosphorylation sites at 41-43 and 119-121; and tyrosine kinase site at 95-102. Nucleotide sequence encoding a rodent IL-174 polypeptide, eg, mouse, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 15 and 16.

CGG CAC AGG CGG CAC AAA GCC CGG AGA GTG GCT GA? GTG GAG CTC TGC 48 Arg His Arg Arg His Lys Wing Arg Arg Val Wing Glu Val Glu Leu Cys 1 5 10 15 ATC TGT ATC CCC CCC AGA GCC TCT GAG CCA CAC CC? DC? CGC AGA ATC 96 He Cys He Pro Pro Arg Wing Ser Glu Pro His Pro Pro A.rg Arg He 20 25 30 CTG CAG GGC CAG CAA GGA TGG CCT CTC AAC AGC AGG GCC ATC TCT CCT 144 Leu Gln Gly Gln Gln Gly Trp Pro Leu Asn Ser Arg Wing He Ser Pro 35 40 _ 45 TGG AGC TAT GAG TTG GAC AGG GAC TTG AAT CGC GTC CCC CAG GAC TGG 192 Trp Ser Tyr Glu Leu Asp Arg Aep Leu Asn Arg Val Pro Gln Asp Trp 50 55 60 TAC CAC GCT CGA TGC CTG TGC CCA CAC TGC GTC ACG CTA CAG ACÁ GGC 240 Tyr His Wing Arg Cys Leu Cys Pro His Cys Val Thr Leu Gln Thr Gly 65 70 75 80 TCC CAC ATG GAC CCG CTG GGC AAC TCC GTC CCA CTT TAC CAC AAC CAG 288 Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn Gln 85 90 95 ACG GTC TTC CTG CGG CCA TGC ATG GCG AGG AAG GTA CCC ATC GCC 336 Thr Val Phe Tyr Arg Arg Pro Cys Met Ala Arg Lys val Pro lie Wing 100 105 110 GCT ACT GCT TGG AGC GCA GGT CTA CCG AGT CTC CTT GGC TTG TGT GTG 384 Wing Thr Wing Trp Ser Wing Gly Leu Pro Ser Leu Leu Gly Leu Cys Val 115 120 125 TGT GCG GCC CCG GGT CAT GGC TTA GTC ATG CTC ACC ATC TGC CTG AGG 432 Cys Wing Wing Pro Gly His Gly Leu Val Met Leu Thr He Cys Leu Arg 130 135 140 TGAATGCCGG GTGGG GAGA GGGCCAGGTG TACATCACCT GCC ATGCGG GCCGGGTTCA 92 AGCCTGCAAA GCCTACCTGA AGCAGCAGGT CCCGGGACAG GATGGAGACT TGGGGAGAAA '552 TCTGACTTTT GCACTTTTTG GAGCATTTTG GGAAGAGCAG GTTCGCTTGT GCTGTAGAGA 612 TGCTGTTG 620 RHRRHKARRVAEVELCICIPPF? SEPHPPRRILQGQQGWPLNSRAISPWSYELDRDLNRVPQDWYHARC T rPHrvTT.nTC.?HMDPLGNSVPLYHMQTVFYRRPCMA.RKVPI/?TA SAGLPSLLGLCVCAAPGHGLVM Supplemental nucleotide sequence encoding a rodent IL-174 polypeptide, eg, mouse, and predicted amino acid sequence. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 17 and 18 atg tac cag gct gtt cica ttc ttg gca atg ate gtg gga acc fall acc 48 Met Tyr Gln Ala Val Ala Phe Leu Ala Met He Vil Gly Thr His Thr -15 -10 -5 -1 gtc age ttg cgg ate cag gag ggc tgc acit falls ttg eco age tgc tgc 96 Val Ser Leu Arg He Gln Glu Gly Cys Ser His Leu Pro Ser Cys Cys 1 5 lú 15 ecc age aaa gag caa gaa ecc ceg gag gag tgg ctg aag tgg age tet 144 Pro Ser Lys Glu Gin Pro Pro Clu Glu Trp Leu Lys Trp Ser Ser 20 25 30 gca tet gtg tec ecc cea gag ect ctg age falls falls falls gca gaa 192 Wing Ser Val Ser Pro Pro Glu Pro Leu Ser His Tnr His His Wing Glu 35 40 45 tec tgc agg gcc age aag gat ggc ecc etc aac age agg gcc ate tet 240 Ser Cys Arg Wing Being Lys Asp Gly Pro Leu A.sn Being Arg Wing Being 50 55 60 ect tgg age tat gag ttg gac agg gac ttg aat cgg gtc ecc cag gac 288 Pro Trp Ser Tyr Glu Leu Asp Arg A.sp Leu Asn Arg Val Pro Gln Asp 65 70 75 80 ctg tac falls gct cga tgc ctg tgc cea falls tgc gtc age cta cag ac 336 Leu Tyr His Wing Arg Cvs Leu Cys Pro His Cys Val Ser Leu Gln Thr 85"90 95 ggc tec falls atg gac ceg ceg ggc aac tech gtc cea ett tac falls aa c 384 Gly Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn 100 105 110 cag acg gtc ttc tac cgg cgg cg cea tgc cat ggt gag gaa ggt acc cat 432 Gln Thr Val Phe Tyr Arg Arg Pro Cys His Gly Glu Glu Gly Thr His 115 120 125 cgc cgc tac tgc ttg gag cgc agg etc tac cga gtc tec ttg gct tgt 480 Arg Arg Tyr Cys Leu Glu Arg Arg Leu Tyr Arg Val Ser Leu Wing Cys 130 135 140 gtg tgt gtg cgg ecc cgg gtc atg gct tagtcatgc.t caccacctgc 527 Val Cys Val Arg Pro Arg Val Met Ala 145 150 ctgaggctga tgcccggttg gyagagaggg ccaggtgtac aatcaccttg ccaatgcggg 587 gccctccaaa ccgggttcaa gccctacctg aagcagcayg ctcccgggac aagatggagg 647 acttggggag aaactctgac ttttgcactt tttggaagca cttttgggaa ggagcaggtt 707 ccgcttgtgc tgctagagga tgctgttgtg gcatttctac tcaggaacgg actccaaagg 767 cctgctgacc ctggaagcca tactcctggc tcctttcccc tgaatccccc aactcctggc 827 acaggcactt tctccacctc tccccctttg ccttttgttg tgtttgtttg tgcatgccaa 887 ctctgcgtgc agccaggtgt aattgccttg aaggatggtt ctgaggtgaa agctgttatc 947 gaaagtgaag agatttatec aaataaacat ctgtgttt 985 MYQAV? FLAMIVGTHTVSLRIQEGCS; HLPSCCPSKEQEPPEEWL WSS? SVSPPEPLSHTHHAESCRAS Important predicted reasons include, for example, cAMP PK sites at 29-32 and 61-64; Ca phosphorylation sites at 18-20, 53-55 and 67-69; Myristoyzalyl site at 123-127; N-glycosylation site at 112-114; and phosphorylation sites in 29, 31, 51, 53, 61, 64, 139 and 141; and PKC phosphorylation sites in 2-4, 49-51 and 127-129.

Table 4: nucleotide sequence encoding a primate IL-171, eg, human, under the IUPAC code. In addition, you can use complementary nucleic acid sequences for many purposes. SEC ID NO: 19: GACACGGATG AGGACCGCTA TCCACAGAAG CTGGCCTTCG CCGAGTGCCT GTGCAGAGGC 60 TGTATCCATG CACGGACGGG CCGCGAGACA GCTGCGCTCA ACTCCGTGCG GCTGCTCCAG 120 AGCCTGCTGG TGCTGCGCCG CCGGCCCTGC TCCCGCGACG GCTCGGGGCT CCCCACACCT 180 GGGGCCTTTC CCTTCCACAC CGAGTTC? TC CACGTCCCCG TCGGCTGCAC CTGCGTGCTG 240 CCCCGTTC A GTGTGACCGC CAAGGCCGTG GGGCCCTTAG NTGACACCGT GTGCTCCCCA 300 GAGGGACCCC TATTTATGGG AATTATGGTA TTATATGCTT CCCACATACT TGGGGCTGGC 360 ATCCCGNGCT GAGACAGCCC CCTGTTCTAT TCAGCTATAT GGGGAGAAGA GTAGACTTTC 420 AGCTAAGTGA AAAGTGNAAC GTGCTGACTG TCTGCTGTCG TNCTACTNAT GCTAGCCCGA 480 GTGTTCACTC TGAGCCTGTT AAATATAGGC GGTTATGTAC C 521 SEQ ID NO: 20 and 21 are PATENTIN translatable cDNA and polypeptide sequences: GAC ACG GAT GAG GAC CGC TAT CCA CAG AAG CTG GCC TTC GCC GAG TGC 48 Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Wing Phe Wing Glu Cys 1 5 10 15 CTG TGC AGA GGC TGT ATC GAT GCA CGG ACG GGC CGC GAG ACÁ GCT GCG 96 Leu Cys Arg Gly Cys He Asp Wing Arg Thr Gly Arg Glu Thr Wing Wing 20 25 30 CTC AAC TCC GTG CGG CTG CTC CAG AGC CTG CTG GTG CTG CGC CGC CGG 14 Leu Asn Ser Val Arg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg Arg 35 40 45 CCC TGC TCC CGC GAC GGC TCG GGG CTC CCC AC CCT GGG GCC TTT GCC 192 Pro Cys Ser Arg Asp Gly Ser Gly Leu Pro Thr Pro Gly Wing Phe Wing 50 55 60 TTC CAC ACC GAG TTC ATC CAC GTC CCC GTC GGC TGC ACC TGC GTG CTG 240 Phe His Thr Glu Phe He His Val Pro Val Gly Cys Thr Cys Val Leu 65 70 75 80 CCC CGT TCA AGT GTG ACC GCC AAG GCC GTG GGG CCC TTA GnT GAC ACC 288 Pro Arg Ser Ser Val Thr Wing Lys Wing Val Gly Pro Leu Xaa Asp Thr 85 90 95 GTG TGC TCC CCA GAG GGA CCC CTA TTT ATG GGA ATT ATG GTA TTA TAT 336 Val Cys Ser Pro Glu Gly Pro Leu Phe Met Gly He Met Val Leu Tyr 100 105 110 GCT TCC CAC ATA CTT GGG GCT GGC ATC CCG nGC TGAGACAGCC CCCTGTTCTA 389 Ala Ser His He Leu Gly Ala Gly He Pro Xaa 115 120 TTCAGCTATA TGGGGAGAAG AGTAGACTTT CAGCTAAGTG AAAAGTGCAA CGTGCTGACT 449 GTCTGCTGTC GTCCTACTCA TCCTAGCCCG AGTGTTCACT CTGA.GCCTGT TAAATATAGG 509 CGGTTATGTA CC 521 DTDEDRYPQKLAFA.ECLCRGCI.J'.RTGRETAA NSVRLLQSLLVLRRRPCSRDGSGLPTPGAFAFHTEFI HVPVGCTCVLFRSSVTAKAVGPLXDTVCSPEGPLFMGIMVLYASHILGAGIPX Supplementary nucleotide sequence encoding a primate IL-171, eg, human. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 22 and 23. gtgtggcctc agotataaga scggctgctg ccaggtgca ggccaggtgc acctgtggga 60 ttgccgccag gtgtgcaggc cgctccaagc ccagcctgcc ccgctgccgc cace atg 117 Met acg etc etc ecc ggc etc ctg ttt ctg acc tgg ctg falls here tgc ctg 165 Thr Leu Leu Pro Gly Leu Leu Phe Leu Thr Trp Leu His Thr Cys Leu -15 -io- -5 -1 gcc falls cat gac ecc tec etc. agg ggg drops ecc falls agt falls ggt acc 213 Ala His His Asp Pro Ser Leu A.rg Gly Hie Pro His Ser His Gly Thr 1 5 10 15 cea falls tgc tac teg gct gag gaa ctg ecc etc ggc cag gcc ecc cea 261 Pro His Cys Tyr Ser Wing Glu Glu Leu Pro Leu Gly Gln Wing Pro Pro 20 25 30 ctg falls ctg gct cg ggt gcc aag tgg ggg cag gct ttg ect gta gcc 309 His Leu Leu Wing Arg Gly Wing Lys Trp Gly Gln Wing Leu Pro Val Wing 35 40 45 ctg gtg tec age ctg gag gca gca age falls agg ggg agg falls gag agg 357 Leu Val Ser Ser Leu Glu Wing Wing His Arg Glv Arg His Glu A.rg 50 55 60 c.cc tea gct acg acc fall tgc ceg gtg ctg cgg ceg g ~ ag gag gtg ttg 405 Pro Ser Ala Thr Thr Gin Cys Pro Val Leu Arg Pro Glu 'Glu Val Leu 65 70 75 80 geg gca gac acc falls cag cgc tec ate tea cec tgg aga tac cgt gtg 453 Glu Wing Asp Thr His Gln Arg Ser He Ser Pro Trp Arg Tyr Arg Val 85 90 95 gac acg gat gag gac cgc tat cea cag aag ctg gcc: tcc gcc gag tgc 501 Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Wing Phe Wing Glu Cys 100 105 110 ctg tgc aga ggc tgt ate gat gca cgg acg ggc cgc gag here gct gcg 549 5 Leu Cys Arg Gly Cys He Asp Ala Arg Thr Gly Arg Glu Thr Ala Ala 115 120 125 etc aac tec gtg cgg ctg etc cag age ctg ctg gtg ctg cgc cgc cgg 597 Leu Asn Ser Val Axg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg Arg 130 135 140 ecc tgc tec cgc gac ggc teg ggg etc ecc here ect ggg gcc ttt gcc 645 Pro Cys Ser A.rg Asp Gly Ser Gly Leu Pro Thr Pro Gly Wing Phe Wing 145 150 155 160 ttc falls acc gag ttc ate drops gtc ecc gtc ggc tgc acc tgc gtg ctg 693 Phe His Thr Glu Phe He His Val Pro Val Gly Cys Thr Cys Val Leu 165 170 175 10 ecc cgt tea gtg tgaccgccga ggccgtgggg cccctagact ggacacgtgt 745 Pro Arg Ser Val 180 gctccccaga gggcaccccc tatttatgtg tatttattgg tatttatatg cctcccc caa 805 cactaccctt ggggtctggg cattccccgt cagcccccca gtctggagga ctgttctcct 865 catctccagc ctcagtagtt gggggtagaa ggagetcage acctcttcca gcccttaaag 925 ctgcagaaaa ggtgtcacac ggctgcctgt accttggcte cctgtcctgc tcccggcttc 985 ccttacccta tcactggcct caggcccccg caggctgcct cttcccaacc tccttggaag 1045 '^ tacccctgtt tettaaacaa ttatttaagt gtacgtgtat tattaaactg atgaacacat 1105 ce 1107 MTLLPGLLFLTWLHTCLAJ-IHDPSLRGFPHSHGTPHCYSAEELPLGQ? PPHLLARGAKWGQALPVALVSS LEAASHRGRHERPSAT ^ QCPVLRPEEVLEADTHQRSISPWRYR ^ / DTDEDRYPQKLAFAECLCRGCIDAR TGRET.AAiNSVRLLQSLLVLRRRPCSRDGSGLPTPGAFAFHTLFIHVPVGCTCVLPRSV Table 5: nucleotide sequence encoding a primate IL-175 sequence, eg, human, under the IUPAC code. In addition, you can use complementary nucleic acid sequences for many purposes. SEC ID NO: 24: GAGAAAGAGC TTCCTGCACA AAGTAAGCCA CCAGCGCAAC ATGACAGTGA AGACCCTGCA 60 TGGCCCAGCC ATGGTCAAGT ACTTGCTGCT GTCGATATTG GGGCTTGCCT TTCTGAGTGA 120 GGCGGCAGCT CGGAAAATCC CCAAAGTAGG ACATACTTTT TTCCAAAAGC CTGAGAGTTG 180 CCCGCCTGTG CCAGGAGGTA GTATGAAGCT TGACATTGGC ATC? TCAATG AAAACCAGCG 240 CGTTTCCATG TCACGTAACA TCGAGAGCCG CTCCACCTCC CCCTGGAATT ACACTGTCAC 300 TTGGGACCCC AACCGGTACC CCTCGAAGTT GTACAGGCCC AAGTGTAGGA ACTTGGGCTG 360 TATCAATGCT CAAGGAAAGG AAGACATCTN CATGAATTCC GTC 403 SEQ ID NO: 25 and 26 are PATENTIN translatable polypeptide and cDNA sequences. Predicted signal decomposition site indicated, but there may be some debris on either side; putative glycosylation site at residues 53-55: GAGAAAGAGC TTCCTGCACA AAGTAAGCCA CCAGCGCAAC ATGAC? GTGA AGACCCTGCA 60 TGGCCCAGCC ATG GTC AAG TAC TTG CTG CTG TCG ATA TTG GGG CTT GCC 109 Met Val Lys Tvr Leu Leu Leu Ser He Leu Gly Leu Ala -20 -. -15 -10 TTT CTG AGT GAG GCG GCA GCT CGG AAA ATC CCC AAA GTA GGA CAT ACT 157 Phe Leu Ser Glu Wing Wing Arg Lvs He Pro Lys Val Gly His Thr -5 1 5 TTT TTC CAA AAG CCT GAG AGT TGC CCG CCT GTG CCA GGA GGT AGT ATG 205 Phe Phe Gln Lys Pro Glu Ser Cys Pro Pro Val Pro Gly Gly Ser Met 10 15 20 25 AAG CTT GAC ATT GGC ATC ATC AAT GAA AAC CAG CGC GTT TCC ATG TCA 253 Lys Leu Asp He Gly He He Asn Glu Asn Gln Arg Val Ser Met Ser 30 35 40 CGT AAC ATC GAG AGC CGC TCC ACC TCC CCC TGG AAT TAC ACT GTC ACT 301 Arg Asn He Glu Ser Are Ser Thr Ser Pro Trp Asn Tyr Thr Val Thr 45 50 55 TGG GAC CCC AAC CGG TAC CCC TCG AAG TTG TAC AGG CCC AAG TGT AGG 349 Trp Asp Pro Asn Arg Tyr Pro Ser Lys Leu Tyr Arg Pro Lys Cys Arg 60 65 70 AAC TTG GGC TGT ATC A? T GCT CAA GG? AAG GAA GAC ATC TCC ATG AAT 397 Asn Leu Gly Cys He Asn Wing Gln Gly Lys Glu Asp He Ser Met Asn 75 80 85 TCC GTC 40 Ser Val 90 MVKYLLLSILGLAFLS? AAARKIP VGHTFFQKPESCPPVPGCeMKLDtGIINENORVSMSRNIESRST SP NYTVTWDPNRYP? KLYRPKCPJÍLGCINAQGKEDIXK? 'SV Particularly interesting segments include, for example, those that start or end with argl; cys17; pro18; pro19; val20; thr49; Ser50; arg69; pro70; and the end of the sequence available. Table 6: nucleotide sequence encoding a primate IL-176, eg, human. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 27 and 28: te gtg ceg tat ett ttt aaa aaa att att ett falls ttt ttt gcc tec 47 Val Pro Tyr Leu Phe Lys Lys He He Leu His Phe Phe Wing Being 1 5 10 15 tat tac ttg tta ggg aga ecc aat ggt age ttt att ect tgg gga tac 95 Tyr Tyr Leu Leu Gly A.rg Pro Asn Gly Ser Phe Tje Pro Trp Gly Tyr 20 25 30 ata gta aat act tea tta aag teg agt here gaa ttt gat gaa aag tgt 143 He Val Asn Thr Ser Leu Lys Ser Ser Thr Glu Phe Asp Glu Lys Cys 35 40 45 gga tgt gtg gga tgt act gcc gcc ttc aga agt cea falls act gcc tgg 191 Gly Cys Val Gly Cys Thr Ala Ala Phe Arg Ser Pro His Thr Ala Trp 50 55 60 agg gag aga act gct tat tea ctg att aag cat ttg ctg tgt acc 239 Arg Glu Arg Thr Wing Val Tyr Ser Leu He Lys His Leu Leu Cys Thr 65 70 75 aac tac ttt tea tgt ett ate att att etc tie here gtc att 281 Asn Tyr Phe Ser Cys Leu He Leu He Leu He Thr Val I 80 85 90 aaaaacccca tgatatttta gaaatctgag aaagagataa agtggtttgc tcaaggttat 341 agaacagact accatgtgtt gtatttcaga ttttaattca tgtttgtctg attttaagtt 401 ttgttcgctt gccagggtac cccacanaaa tgccaggcag ggcattttca tgatgcactt 461 gagatacctg aaatgacagg gtagcatcac aectgagagg ggtaaaggat gggaacctac 521 cttccatggc cgctgcttgg cagtctcttg ctgeatgeta gcagagccac tgtatatgtg 581 ccgaggctct gagaattaac tgcttaaaga actgccttct. ggagggagaa gagcacaaga 641 tcacaattaa ccatatacac atcttactgt gcgaggtcat tgagcaatac aggagggatt 701 ttatacattt tageaactat cttcaaaacc tgagctatag ttgtattctg cccccttcct 761 ctgggcaaaa gtgtaaaagt ttg 784 VPYLFKKIILHFFASYYLLGRPNGSFIPWGYIVNTSLKSSTEFDEKCGCVGCTAAFRSPHTA RER TAVYSLIKHLLCTNYFSCLILILITVI Nucleotide sequence encoding a primate IL-177, eg, human. In addition, you can use complementary nucleic acid sequences for many purposes. SEQ ID NO: 29 and 30: gtg act gta ttg tgg gga cag gaa gca cac att ecc atg tgg ate act 48 Val Thr Val Lea Trp Gly Gln Gllu Ala Gln He Pro Mat Trp He Thr 1 5 10 15 agg aga gat aat aag tgg ggt cat ttc acc ect tgg tec ect gct tec 96 Arg Arg Asp Asn Lys Trp Gly His Phe Thr Pro Trp Ser Pro Wing Ser 20 25 30 aga ecc aaa gag gcc tac atg gca ttg tgc ttc ett ett agt tgt agg 144 Arg Pro Lys Glu Ala Tyr Me.t Ala Leu Cys rhe Leu Leu Ser Cys Arg 35 40 45 agg tgt gag ata caa tea ttt gcc tet gac ttt gag ggt tgg tec 189 Arg Cys Glu He Gln Ser Phe Wing Being Asp Phe Glu Gly Trp Ser 50 55 60 tagcatgccc ctgaccagta gccccttaaa tacttcattg atatggaagg tctctgaatc 249 ttcgtgggct taatctacca ctctctgaag ttcttatgtc tttcaaaggc ctctaaaatc 309 tctgccatgt cttgctcatc casttgttag catgacgtca ttgatacagt ggactttgga 369 atctaagtgg ggagacactg gtaagtgaec aattacttca cctgtggtgt gcaagccaga., 429 tcaggaagcc tctacctgca cgacaacaca t 460 VTVÍ, GQEAQIPMWITRRDNK GHFTPWSPASRPKEAY! ALCFLLSCRRCEIQ? FASDFEGVÍS Table 7: alignment of several members of the CTLA-8 family / IL-170. The rat CTLA-8 sequence is SEQ ID NO: 31 (see GB L13839; 293329/30; the mouse CTLA-8 sequence is SEQ ID NO: 32 (see GB 1469917/8), human CTLA-8 is SEQ ID NO: 32 NO: 33 (see GB U32659; 115222/3); and Herpes Saimiri virus ORF 13 is SEQ ID NO: 34 (see GB Y13183; 2370235): Multiple sequence alignment CLUSTAL X (1.64b).

IL-74 Mu MYQAVAFLAMIVGTHTVSLRI QEGCSHLPSCCPSKEQEPPEEWLKWS IL-74 ~~ Hu MYQWAFLAMVMCTHTY S --- HWPSCCPSKGQDTSEELLRWS IL-72 ~ Hu MDWPHNLLFLLTISIFLGLGQPRSPKSKRKGQGRPGPLVPGPHQVPLDLVSRMK IL-72 ~ Mu MD PHSLLFLLAISIFLAP? HPRNTKGKRKGQGRPSPLAPGPHQVPLDLVSRVK IL-73_Mu -M GTLV MLLVGFL ALAPGRAAGALRT - GRRP - ARPRDCADRPEE LEQLYGRLA IL-73 ~ Hu MVAGFLLALPPS AAGAPRA - GRRP - ARPRGCADRPEELLEQLYGRLA IL-17 ~ Hu --MTPGKTSLVSLLLLLSLEAIVKAGITIP RNPGCPNSEDKNFPRTVMVNL TT 17 ~ Hs --MTFRKT? LV-LLLLLSIDCIVKSEITSA QTPRCLAAMN-SFPRSVMVTL ? L-17 ~ Rt MCLMLLLLLNLEATVK? AVLIP QSSVCP AEANNFLQ VKV L IL-17 ~ Mu MLLLLLSLAATVKAAAIIP QSSACPNTEAKDFLQ VKVNL IL-75 ~ Hu MVKYLLLSILGLAFLSEAAARKIPKVGHTFFQKPESCPPVPGGSMKLDIGI1N IL-7I! HU MTLLPCLLFLTWLHTCLAHHDPSLRGHPHSHGTPHCYSAEELPLGQAPPHLLARGAK GQ IL- -74 Mu S ASVSPP-EPLSHTHHAES CRASKD-GPLNSRAISP SYELDRDLNRV IL- -74 Hu t VPVPPL-EPARPNRHPES CRASED-GPLNSRAISPWRYELDRDLNRL IL- -72 Hu P-YARI1EEYER IEEMVAQLPJ > ISSELAQ-RKCEVNLQL I > ISNKRSLSPWGYSINHDPSRI IL- -72 Mu P-YARMEEYERNLGEMVAQLR S? EPAK-KKCEV LOL LSNKRSLSPWGYSINHDPSRI IL- -73 Mu? GVLSAFHHTLQIßPR-EQARNASCPAGGRAADRRFR-PPTNLRSVSPWAYRISYDPARF IL- -73"Hu AGVLSAFHHTLQIiGPR-EQARIJASCPAGGRP DRRFR-PPTNLRSVSP AYRISYDPARY IL- -17. Hu w IHNRKTNTN P-KRSSDYYNRSTSPWNLHRNEDPERY IL- -17 _Hs S IRNNTSS KRASDYYNRSTSPWTLHRNEDQDRY IL- -17 _Rt K VINSLSSKA SSRRPSDYLNRSTSPWTLSRNEDPDRY IL- -17 Mu K VFNSLGAKV SSRRPSDYLNRSTSPWTLHRNEDPDRY IX- -75 _Hu E N - QRVSMS R - NIESRSTSPWNYTVTWDPNRY IL- -71 _Hu ALPVALVS? LEAASHRGRHERPSATTQCPVLRPEEVLEADTHQRSISPWRYRVDTDEDRY IL-74_MU PQDLYHAPCLCPHCVSLQTGSHMDPLGNSVPLYHNOTVFYRR - PCHGEEGTHRRYCLER IL-74_HU PQDLYHAFCLCPHCVSLQTG5HMDPRGNSELLYHNQTVFYRR - PCHGEKGTHKGYCLER IL-72_HU PVDLPEAGCLCLGCVNPFTM-QEDRSMVSVPVFS-QVPVRRR - LCPPPP - RTGPCRCF IL-72_MU P? DLPE? G.CÚCLGCVNPFTM-OEDRSÍ'ÍVSVPVFS-QVPVRRR-LCPQPP - RPGPCRCR IL-73._Mu PRYLPE? YCLCRGCLTGLYG-EEDFRFR? TPVFS-PAWLRRTATAGAG GRSVY? IL-73_Hu PRYLPEAYCLCRGCLTGLFG-EEDVRFRSAPVYM-PTWLRRTPACAG GRSVYT IL- 17_Hu P? VIWEAKCRHLGCINADGN- -VDYHM SVPIQQEILVLRREPPHCPN SFR IL- 17_Hs PSVIWEA CRYLGCVNADGN ---- VDYHM SVPIQQEILWRKGHQPCPN SFR IL-17_Rt PSVIWEAQCRHQRCVNAEGK- -LDHHM SVLIQQE3 VLKREPEKCPF TFR IL-17_Mu PSVIWE /? QCRHQRCVNAEGK - LDHHMNSVLIQQEILVLKREPESCPF TF R IL- 75_Hu PSE QAQCRNLGCINAQGK- -EDISMNSVPIQQETLWRRKHQGCSV SFG IL-71_Hu PQKLAFAECLCRGCIDARTG-RETAALNSVRLLQSLLVLRRRPCSRDGSGLPTPGAFAFH IL -74_Mu R YR-VSÍ? CVCVRPRVM? ? IL-74_Hu R YR-VSLACVCVRPRVMG IL-72_Hu AV ^ TIAVGCTCIF IL-72_Mu WMETI VGCTCIF IL-73_Mu EHYITIPVGCTCVPEPDKSADSANSSMDK LLLGPADRPAGR IL-73_Hu E YVTIPVGCTCVPEPEKDADSINSSIDKOGAKLLLGPNDAPAGP IL- 17_Hu LEKTL7EÑ / GCTCVTPIVHHVA IL- 17_Hs LEKMJJ / TVGCTCVTPIVHNVD IL-17_Rt VEKMLVGVGCTCVSSIVRHAS - IL-17_Mu VEKMLVCVG TCVASIVRQAA IL-7 5_HU "" LEKVLVTVGCTCVTPVIHHVQ IL-71_Hu TEFXHVPVGCTCVLPRSV *.

Particularly interesting segments include, for example, those corresponding to the segments of IL-172 or IL-175, indicated above, with the other members of the family. The purified polypeptides or proteins are useful for the generation of antibodies by standard methods, as described above. The synthetic peptides or purified protein can be presented to an immune system to generate a specific binding composition, for example, monoclonal or pliclonal antibodies. See, for example, Coligan (1991) Current Protocols in Immunoloqy and Wiley / Greene: and Harlow and Lane (1989) Antibodies: A Laboratory Manual Cold Spring Harbor Press. For example, the specific binding composition could be used for screening an expression library made from a cell line expressing an IL-170 protein. The screening may be standard protein staining expressed on the surface, or by panning. The screening of intracellular expression can also be performed by various staining or immunofluorescence procedures. The binding compositions could be used to purify by affinity, or to classify cells expressing the protein. This invention contemplates the use of isolated DNA or fragments to encode a corresponding biologically active IL-170 protein or polypeptide. In addition, this invention covers isolated or recombinant DNA that encodes a biologically active protein or polypeptide, and that is capable of hybridizing under appropriate conditions to the DNA sequences described herein. Said biologically active polypeptide or protein can be an intact antigen, or fragment, and have an amino acid sequence as described in tables 1-6. In addition, this invention covers the use of isolated or recombinant DNA, or fragments thereof, which encodes proteins that are homologous to an IL-170 protein or that were isolated using cDNA encoding an IL-170 protein as a probe. The isolated DNA may have the respective regulatory sequences on the 5 'and 3' flanks, for example, promoters, enhancers, poly-A addition signals and others. An "isolated" nucleic acid is a nucleic acid, for example, a RNA, DNA or a mixed polymer, which is substantially separated from other components that naturally accompany a natural sequence, for example, robosomes, polymerases and flanking genomic sequences of the species of origin. The term encompasses a nucleic acid sequence that has been removed from its natural environment, and includes recombinant or cloned DNA isolates and chemically synthesized analogs or biologically analogs synthesized by heterologous systems. A substantially pure molecule includes isolated forms of the molecule. Alternatively, a purified species can be separated from host components of a recombinant expression system. The homology size of said nucleic acid will typically be less than large vectors, eg, less than ten kB, typically less than several kB, and preferably in the range 2-6 kB.

An isolated nucleic acid will generally be a homogeneous composition of molecules, but, in some embodiments, will contain less heterogeneity. This heterogeneity is typically found at polymer ends or non-critical portions for a desired biological function or activity. A "recombinant" nucleic acid is defined either by its production method or by its structure. With reference to its production method, for example, a product made by a process, the process is the use of recombinant nucleic acid techniques, for example, which involve human intervention in the nucleotide sequence, typically selection or production. Alternatively, it may be a nucleic acid made by generating a sequence comprising the fusion of two fragments that are not naturally contiguous with each other, but is intended to exclude products of nature, for example, natural mutants. Thus, for example, products made by transforming cells with any non-natural vector are encompassed, as are nucleic acids comprising sequence derived using any synthetic oligonucleotide process. This is often done to replace a codon with a redundant codon encoding the same amino acid or a conservative, while typically entering or removing a sequence recognition site. Alternatively, it is performed by joining nucleic acid segments of desired functions, to generate a single genetic entity comprising a desired combination of functions that are not found in the commonly available natural forms. Restriction enzyme recognition sites are often the target of such artificial manipulations, but other site-specific targets, for example, promoters, DNA replication sites, regulatory sequences, control sequences, or other useful features, may be incorporated by design. A similar concept is proposed for a recombinant, eg, fusion polypeptide. Synthetic nucleic acids are specifically included which, by redundancy of genetic code, encode polypeptides similar to fragments of these antigens, and fusions of sequences of several different species variants. A significant "fragment" in a nucleic acid context is a contiguous segment of at least about 17 nucleotides, in general, at least 20 nucleotides, more generally, at least 23 nucleotides. Commonly at least 26 nucleotides, more commonly at least 29 nucleotides, often at least 32 nucleotides, more often at least 35 nucleotides, typically at least 38 nucleotides, more typically at least 41 nucleotides, usually at least minus 44 nucleotides, more usually at least 47 nucleotides, preferably at least 50 nucleotides, more preferably at least 53 nucleotides, and in particularly preferred embodiments, will be at least 56 or more nucleotides. Such fragments can have terms in any location, but especially in the boundaries between structural domain.In other embodiments, the invention provides polynucleotides (or polypeptides) comprising a plurality of distinct, for example, non-overlapping segments of the specified length. Typically, the plurality will be at least two, more usually at least three, and preferably 5, 7 or even more. While length minima are provided, longer lengths, of various sizes, may be appropriate, for example, one of length 7, and two of length 12. A DNA encoding an IL-170 protein will be particularly useful for identifying species of genes, mRNA and cDNA that code for related proteins or homologs, as well as DNAs that code for homologous proteins of different species. There are equal homologs in other species, including primates. Several CTLA-8 proteins should be homologous and are encompassed herein. However, even proteins that have a more distant evolution relationship to the antigen can be easily isolated under appropriate conditions, using these sequence if they are sufficiently homologous. Primate CTLA-8 protein proteins are of particular interest. This invention also covers recombinant DNA molecules and fragments having a DNA sequence identical or highly homologous to the isolated DNAs that are disclosed herein. In particular, the sequences will often be operably linked to DNA segments that control the transcription, translation and replication of DNA. Alteratively, recombinant clones derived from genomic sequences, for example, containing neutrons, will be useful for transgenic studies, including, for example, transgenic cells and organisms, and for gene therapy. See, for example, Goodnow (1992) "Transgenic Animáis" e Roitt (ed.) Encyclopedia of Immunoloqy Academic Press, San Diego, pages 1502 - 1504; Travis (1992) Science 256: 1392-1394; Kuhn et al. (1991) Science 254: 707-710; Capecchi (1989) Science 244: 1288, Robertson (ed. 1987) Teratocarcinomas and Embryonic Stem Cells: A Practical Approach IRL Press, Oxford; Rosenberg (1992) J. Clinical Qncology 10: 180-199; and Cournoyer and Caskey (1993) Ann. Rev. Immunol. 11: 297-329. The homologous nucleic acid sequences, when compared, exhibit significant similarity. Standards for nucleic acid homology are either measured for homology generally used in the art by comparison of sequences, or are based on hybridization conditions. Hybridization conditions are described in greater detail below. Substantial homology in the context of nucleic acid sequence comparison means that the segments, or their complementary strands, when aligned optically, with appropriate insertions or deletions of nucleotides, in at least about 50% of the nucleotides, in general at least less 56% more generally at least 59%, commonly at least 65%, often at least 68%, more often at least 71%, typically at least 74%, more typically at least 77% , usually at least 80%, more usually at least about 85%, preferably at least about 90%, more preferably at least about 95 to 98% or more, and in particular embodiments, as high as about 99 % or more of the nucleotides. Alternatively, substantial homology exists when the segments will hybridize under selective hybridization conditions, to a strand, or to its complement, typically using a sequence derived from Table 2, 3, or 6. Typically, selective hybridization will occur when there is at least about 55% homology over a stretch of at least about 14 nucleotides, preferably at least about 65%, more preferably at least about 75%, and most preferably at least about 90%. See Kanehisa (1984) Nuc. Acids Res. 12: 203-213. The length of homology comparison, as described, may be on longer stretches, and in certain embodiments, it will be on a stretch of at least about 17 nucleotides, usually at least about 20 nucleotides, more usually at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 40 nucleotides, preferably at least about 50 nucleotides, and more preferably at least about 75 to 100 or more nucleotides. Severe conditions, with reference to homology in the hybridization context, will be severe combined conditions of salt, temperature, organic solvents and other parameters, typically those controlled in hybridization reactions. Severe temperature conditions will usually include temperatures above about 30 ° C, more usually above about 37 ° C, typically above about 45 ° C, more typically above about 55 ° C, preferably above about 65 ° C, and more preferably greater than about 70 ° C. Severe salt conditions will commonly be less than about 1,000 mM, usually less than about 500 mM, more commonly less than about 400 mM, typically less than about 300 mM, preferably less than about 200 mM, and more preferably less than about 200 mM. approximately 150 mM. However, the combination of parameters is much more important than the measurement of any individual parameter. See, for example, Wetmur and Davidson (1968) J. Mol. Biol. 31: 349-370. Hybridization under severe conditions should provide a background of at least 2 times above the background, preferably at least 3-5 or more. Alternatively, for sequence comparison, typically one sequence acts as a reference sequence, to which evaluation sequences are compared. When a sequence comparison algorithm is used, the evaluation and reference sequences are loaded into a computer, subsequence coordinates are designated, if necessary, and the parameters of the sequence algorithm program are designated. The sequence comparison algorithm then calculates the percentage of sequence identity for the evaluation sequence (s) in relation to the reference sequence, based on the designated program parameters. The optical alignment of sequences for comparison can be conducted, for example, by the local homology algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2: 482, by the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48: 443, by the search for similarity method of Pearson and Lipman (1988) Proc. Nat'l Acad. Sci. USA 85: 2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsisn Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wl), or by visual inspection (see generally Ausubel and others, above). An example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment of a group of related sequences, using alignments in pairs, progressive, to show the relationship and percentage of sequence identity. In addition, diagram a tree or dendrogram that shows the grouping relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng and Doolittle (1987) J. Mol. Evol. 35: 351-360. The method used is similar to the method described by Higgins and Sharp (1989) CABIOS 5: 151-153. The program can align up to 300 sequences, each with a maximum length of 5,000 nucleotides or amino acids. The multiple alignment procedure begins with the alignment in the form of pairs of the two most similar sequences, producing a cluster of two aligned sequences. This cluster, then it is aligned to the next most related sequence or cluster of aligned sequences. Two clusters of sequences are aligned by a simple extension of the alignment in the form of pairs of two individual sequences. The final alignment is achieved by a series of alignments in the form of pairs, progressive. The program is executed by designating specific sequences and their coordinates of amino acids or nucleotides for regions of sequence comparison, and designating the parameters of the program. For example, a reference sequence can be compared to other evaluation sequences, to determine the percentage relationship of sequence identity using the following parameters: space weight default (3.00), weight space length default (0.10), and spaces heavy ends. Another example of an algorithm that is suitable for determining the percent sequence identity and sequence similarity is the BLAST algorithm, which is described by Altschul et al. (1990) J. Mol. Biol. 215: 403-410. Software for performing BLAST analyzes is publicly available through the National Center for Biotechnology Information (http: www.ncbi.nlm.nih.gov/). This algorithm involves first identifying pairs of high-ranking sequences (HSPs), identifying short words of length W is the sequence of questions, which either comply with, or satisfy, some of the positive-value T-threshold classifications when aligned with a word of the same length in a database sequence. T is referred to as the threshold of classification of neighboring words (Altschul et al., Above). These initial neighbor word hits act as seeds for the initiation of searches to find longer HSPs that contain them. Word hits then extend in both directions along each sequence, as much as the cumulative alignment ranking can be increased. The extension of word hits in each direction stops when: the cumulative alignment classification overflows by the amount of X of its maximum value achieved; the cumulative classification goes to zero or lower, due to the accumulation of one or more alignments of negative classification residues; or the end of any sequence is achieved. The parameters of the BLAST algorithm W, T and X determine the sensitivity and speed of the alignment. The BLAST program uses as defaults a word length (W) of 11, the BLOSUM62 classification matrix alignments (see Henikoff and Henikoff (1989) Proc. Nat'l. Acad. Sci. USA 89: 10915) (B) 50, expectation (E) of 10, M = 5, N = 4 and a comparison of both strands. In addition to calculating the percentage of sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin and Altschul (1993) Proc. Nat'l. Acad. Sci. USA 90: 5873- 5787). One measure of similarity provided by the BLAST algorithm is the minus sum probability (P (N)), which provides an indication of the probability by which a binding between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the likelihood of a minor sum in a comparison of the assay nucleic acid to the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and with more preference, less than about 0.001. Another indication that two polypeptide nucleic acid sequences are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically reactive cross-linked with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, when the two polypeptides differ only by conservative substitutions. Another indication that two substantially identical nucleic acid sequences is that the two molecules hybridize to each other under severe conditions, as described below. CTLa-8 proteins from other mammalian species can be cloned and isolated by cross-species hybridization of related species, eg, human, as described in tables 1-7. The homology may be relatively low between distantly related species, and on the other hand, it is advisable to hybridize relatively closely related species. Alternatively, the preparation of an antibody preparation that exhibits less species specificity may be useful in expression cloning approaches. lll. Purified IL-170 protein The predicted sequence of primate, eg, human, and rodent polypeptide IL-173, eg, mouse, is shown in table 2. Likewise, in table 3, IL-174 sequence is provided. primate, for example, human, and is assigned SEQ ID NO: 14. In addition, a rodent IL-174, for example murine, is described in Table 3. Peptide sequences allow the preparation of peptides to generate antibodies to recognize said segments . As used herein, the terms "primate IL-170 protein" and "rodent IL-170 protein" will encompass, when used in a protein context, a protein having designated amino acid sequences shown in the tables 1-7, or a significant fragment of said protein. It also refers to a polypeptide derived from primate or rodent that exhibits similar biological function, or interacts with specific binding components of IL-170 protein. These linker components, for example antibodies, typically bind to an IL-170 protein with high affinity, for example, therefore less than about 100 nM, usually better than 30 nM, preferably better than about 10 nM, and with more preference, to better than about 3 nM. Homologous proteins would be found in species of non-rat or human mammals, for example, mouse, primates and in the herpes virus genome, for example, ORF13. Non-mammalian species should also possess structurally or functionally related genes and proteins. The term "polypeptide" as used herein includes a fragment or segment significant a stretch of amino acid residues of at least about 8 amino acids, in general, at least 10 amino acids, more generally at least 12 amino acids, often at least 14 amino acids, more often at least 16 amino acids, typically at least 18 amino acids, more typically at least 20 amino acids, usually at least 22 amino acids, more usually at least 24 amino acids, preferably at least at least 26 amino acids, more preferably at least 28 amino acids, and in particularly preferred embodiments, at least about 30 or more amino acids. The specific ends of said segment will be in any combination within the protein, preferably encompassing structural domains. The term "binding composition" refers to molecules that bind with specificity to IL-170 protein, for example, in a receptor-ligand-like manner, an antibody-antigen interaction, or compounds, for example, proteins that are specifically associate with IL-170 protein, for example, in a natural physiologically relevant protein-protein interaction, whether covalent. The molecule can be a polymer, or chemical reagent. No hint is represented as to whether the IL-170 protein is either the ligand or the receptor of a ligand-receptor interaction, other than that the interaction exhibits similar specificity, e.g., specific affinity. A functional analog can be a protein with structural modifications, or it can be a completely unrelated molecule, for example, having a molecular form that interacts with the appropriate binding determinants. Proteins can serve as agonists or antagonists of a receptor, see, for example, Goodman, et al. (Eds. 1990) Goodman & Gilman's: The Pharmacological Bases of Therapeutics (8th Ed.), Pergamon Press. The solubility of a polypeptide or fragment depends on the environment and the polypeptide. Many parameters affect the solubility of the polypeptide, including temperature, electrolyte environment, size and molecular characteristics of polypeptide, and nature of the solvent. Typically, the temperature at which the polypeptide is used varies from about 4 ° C to about 65 ° C. Usually the temperature in use is greater than about 18 ° C, and more usually about 22 ° C. For diagnostic purposes, the temperature will usually be about room temperature or hotter, but less than the denaturing temperature of components in the test. For therapeutic purposes, the temperature will usually be body temperature, typically about 37 ° C for humans, although under certain situations the temperature may rise or fall in situ or in vitro.

Electrolytes will usually approximate physiological conditions in situ, but can be modified to greater or lesser ionic strength when convenient. Real ions can be modified, for example, to conform to standard pH regulators in physiological or analytical contexts. The size and structure of the polypeptide should generally be in a substantially stable state, and is usually not a denatured state. The polypeptide may be associated with other polypeptides in a quaternary structure, for example, to confer solubility, or it may be associated with lipids or detergents in a manner that approximates bilayer interactions of natural lipids. The solvent will usually be a biologically compatible pH regulator, of a type used for the preservation of biological activities, and will usually approximate physiological solvent. Usually the solvent will have a neutral pH, typically between about 5 and 10, and preferably about 7.5. In some instances a detergent, typically a mild non-denaturing, eg, CHS or CHAPS, or a sufficiently low concentration will be added so as to avoid significantly disrupting the structural or physiological properties of the antigen. The solubility is reflected by the sedimentation measured in Svedberg units, which are a measure of the sedimentation rate of a molecule under particular conditions. The determination of the sedimentation rate was conventionally carried out in an analytical ultracentrifuge, but is now typically carried out in a standard ultracentrifuge. See Freifelder4 (1982) Physical Biochemistry (2nd ed.). W.H. Freeman; and Cantor and Schimmel (1980) Biophvsical Chemistrv, parts 1-3, W.H. Freeman & Co., San Francisco. As a crude determination, a sample containing a putatively soluble polypeptide is rotated in a standard full-size ultracentrifuge, at about 50K, rpm, for about 10 minutes, and the soluble molecules will remain in the supernatant. A soluble polypeptide or particle will typically be less than about 30S, more typically less than about 15S, usually less than about 10S, more usually less than about 6S, and in particularly preferred embodiments, preferably less than about 4S, and more preferably less. at approximately 3S.

IV. Preparation of IL-170 protein; Mimetics DNA encoding the IL-170 protein or fragments thereof can be obtained by chemical synthesis, by screening cDNA libraries, or by screening libraries prepared from a wide variety of cell lines or tissue samples. This DNA can be expressed in a wide variety of host cells for the synthesis of a full-length protein or fragments which can in turn, for example, be used to generate polyclonal or monoclonal antibodies; for link studies; for the construction and expression of modified molecules; and for structure / function studies. Each antigen or its fragments can be expressed in host cells that are transformed or transfected with appropriate expression vectors. These molecules can be substantially purified to be free of protein or cellular contaminants, which are not those derived from recombinant host, and are therefore particularly useful in pharmaceutical compositions when combined with a pharmaceutically acceptable carrier and / or diluent. The antigen, or portions thereof, can be expressed as fusions with other proteins. Expression vectors are typically self-replicating DNA or RNA constructs containing the desired antigen gene or its fragments, usually operably linked to suitable genetic control elements that are recognized in a suitable host cell. These control elements are capable of effecting expression within a suitable host. The specific type of control elements necessary to effect the expression will depend on the possible host cell used. In general, the genetic control elements may include a prokaryotic promoter system or a eukaryotic promoter expression control system, and typically include a transcription promoter, an optional operator to control the start of transcription, transcription enhancers to raise the level of mRNA expression, a sequence encoding a suitable ribosome binding site, and sequences that terminate transcription and translation. Expression vectors also usually contain an origin of replication that allows the vector to replicate independently of the host cell. The methods for amplifying the number of vector copies are also known; see for example, Kaufman et al. (1985) Molec. and Cell. Biol. 5: 1750-1759. The vectors of this invention contain DNA encoding an IL-170 protein, or a fragment thereof, typically encoding a biologically active polypeptide. The DNA can be under the control of a viral promoter, and can encode a selection marker. This invention further contemplates the use of said expression vectors that are capable of expressing eukaryotic cDNA encoding an IL-170 protein in a prokaryotic or eukaryotic host, where the vector is compatible with the host, and where the eukaryotic cDNA encoding the antigen is inserted into the vector, so that the growth of the host containing the cDNA expresses the vector in question. Usually, expression vectors are designated for stable replication in their host cells, or for amplification to greatly increase the total number of copies of the desirable gene per cell. It is not always necessary to require that an expression vector is replicated in a host cell, for example, it is possible to effect the transient expression of the antigen or its fragments in several hosts using vectors that do not contain an origin of replication that is recognized by the host cell . It is also possible to use vectors that cause the integration of an IL-170 protein gene or its fragments, into the host DNA by recombination, or integrate a promoter that controls the expression of an endogenous gene. Vectors, as used herein, comprise plasmids, viruses, bacteriophages, integrable DNA fragments, and other vehicles that allow the integration of DNA fragments into the host's genome. Expression vectors are specialized vectors that contain genetic control elements that effect the expression of operably linked genes. Plasmids are the most commonly used form of vector, but all other forms of vectors that serve an equivalent function, and which are, or become, known in the art, are suitable for use herein. See, for example, Pouwels et al. (1985 and Supplements) Cloninq Vectors: A Laboratory Manual, Elsevier, N. Y., and Rodriguez et al. (Eds. 1988) Vectors: A Survey of Molecular Cloning Vectors and Their Uses. Buttersworth, Boston, MA. Transformed cells include cells, preferably mammals, that have been transformed or transfected with vectors containing an IL-170 gene, typically constructed using recombinant DNA techniques. The transformed host cells usually express the antigen or its fragments, but for purposes of cloning, amplification and manipulation of their DNA, they do not need to express the protein. This invention also contemplates the cultivation of transformed cells in a nutrient medium, thus allowing the protein to accumulate in the culture. The protein can be recovered, either from the culture or from the culture medium. For purposes of this invention, the DNA sequences are operably linked when functionally related to each other. For example, DNA for a presequence or secretory leader is optionally linked to a polypeptide if it is expressed as a preprotein or participates in the direction of the polypeptide to the cell membrane or in the secretion of the polypeptide. A promoter is operably linked to a coding sequence if it controls the transcription of the polypeptide; A ribosome binding site is operably linked to a coding sequence if it is located to allow translation. Usually, operably linked means contiguous and in reading frame, however, certain genetic elements, such as repressor genes, are not contiguously linked, but still link to operator sequences that in turn control the expression. Suitable host cells include prokaryotes, lower eukaryotes and higher eukaryotes. Prokaryotes include both gram negative and gram positive organisms, for example, E. coli and B. subtilis. Minor eukaryotes include yeasts, for example, S. cerevisiae and Pichia and species of the genus Dictyostelium. Higher eukaryotes include established tissue cell lines, from animal cells, both of non-mammalian origin, eg, insect cells, and birds, and of mammalian origin, eg, human, primate and rodent.The prokaryotic host vector systems include a wide variety of vectors for many different species. As used herein, E. coli and its vectors will be used generically to include equivalent vectors used in other prokaryotes. A representative vector for amplifying DNA is pBR322, or many of its derivatives. Vectors that can be used to express IL-170 proteins or fragments thereof include, but are not limited to, vectors such as those containing the lac promoter (pUC series); trp promoter (pBR322-trp); Ipp promoter (the pIN series); lambda-pP or pR promoters (pOTS); or hybrid promoters such as ptac (pDR540). See Brosius et al. (1988) "Expression Vectors Employing Lambda-, trp-, lac-, and Ipp-derived Promoters", in Rodríguez and Denhardt (eds) Vectors: A Survev of Molecular Cloninq Vectors and Their Uses, Buttersworth, Boston, Chapter 10, pages 205-236. Minor eukaryotes, for example, yeast and Dictyostelium, can be transformed with vectors that encode IL-170 proteins. For purposes of this invention, the most common minor eukaryotic host is the baker's yeast, Saccharomyces cerevisiae. It will be used to generically represent minor eukaryotes, although a number of other strains and species are also available. Yeast vectors typically consist of an origin of replication (unless of the integrating type), a selection gene, a promoter, DNA encoding the desired protein or its fragments, and sequences for translation termination, polyadenylation and termination of transcription. Suitable expression vectors for yeast include constitutive promoters such as 3-phosphoglycerate kinase and various other promoters of glycolytic enzyme genes, or inducible promoters such as the alcohol dehydrogenase 2 promoter or metallothionine promoter. Suitable vectors include derivatives of the following types: low self-replicating copy number (such as the YRp series), high self-replicating copy number (such as the YEp series): integrating types (such as the Ylp series), or mini-chromosomes (such as the YCp series). Higher eukaryotic tissue culture cells are the preferred host cells for expression of the functionally active IL-170 protein. In principle, many cell lines of higher eukaryotic tissue culture can be processed, for example, insect baculovirus expression systems, either from an invertebrate or vertebrate source. However, mammalian cells are preferred, in terms of processing, both in the form of co-translation and post-translation. The transformation or transfection and propagation of said cells has become a routine procedure. Examples of useful cell lines include HeLa cells, Chinese hamster ovary (CHO) cell lines, baby rat kidney cell lines (BRK), insect cell lines, bird cell lines and monkey cell lines (COS). Expression vectors for said cell lines usually include an origin of replication, a promoter, a translation initiation site, RNA plating sites (if genomic DNA is used), a polyaddenylation site, and a transcription termination site. These vectors also usually contain a selection gene or amplification gene. Suitable expression vectors can be plasmids, viruses, or retroviruses carrying promoters derived, for example, from sources such as adenovirus, SV40, parvovirus, vaccinia virus or cytomegalovirus. Representative examples of suitable expression vectors include pCDNAl; pCD; see Okayama et al. (1985) Mol. Cell Biol. 5: 1136-11422; pMCIneo Poly-A, see Thomas et al. (1987) Cell 51: 503-512; and a baculovirus vector such as pAC 373 or pAC 610, see O'Reilly et al. (1992) Baculovirus Expression Vectors: A Laboratory Manual Freeman and Co., CRC Press, Boca Raton, Fia. It will often be desired to express an IL-170 protein polypeptide in a system that provides a specific or defined glycosylation pattern. In this case, the usual pattern will be that provided naturally by the expression system. However, the pattern will be modifiable by exposure of the polypeptide, for example, a non-glycosylated form, or appropriate glycosylating protein introduced into a heterologous expression system. For example, the IL-170 protein gene can be co-transformed with one or more genes encoding mammalian or other glycosylating enzymes. Using this approach, certain patterns of mammalian glycosylation can be achieved or approximated in prokaryotic cells, or others. The IL-170 proteins, or a fragment thereof, can be designed to be phosphatidyl inositol (Pl) linked to a cell membrane, but can be removed from the membranes by treatment with a phosphatidyl inositol decomposition enzyme, for example, phosphatidyl inositol phospholipase-C. This releases the antigen in a biologically active form, and allows the purification by standard procedures of protein chemistry. See, for example, Low (1989) Biochim. Biophvs. Minutes 988: 427-454; Tse et al. (1985) Science 230: 1003-1008; and Brunner et al. (1991) J. Cell Biol. 114: 1275-1283. Now that the IL-170 protein has been characterized, fragments or derivatives thereof can be prepared by conventional processes for the synthesis of peptides. These include processes such as described in Stewart and Young (1984) Solid Phase Peptide Svnthesis. Pierce Chemical Co., Rockford, IL; Bodanszky and Bodanszky (1984) The Practice of Peptide Svnthesis, Springer-Verlag, New York; and Bodanszdy (1984) The Principies of Peptide Svnthesis Springer-Verlag, New York. For example, an azide process, an acid chloride process, an acid anhydride process, a mixed anhydride process, an active ester process (e.g., p-nitrophenyl ester, N-hydroxysuccinimide ester, or cyanomethyl) can be used. ester), a carbodiimidazole process, an oxidative reduction process, or a dicyclohexylcarbodiimide (DCCD) / additive process. The solution phase and solid phase syntheses are both applicable to the above processes. The IL-170 protein, fragments, or derivatives, are suitably prepared according to the above processes as are typically employed in the synthesis of peptides, generally by a so-called step process, which comprises condensing an amino acid to the terminal amino acid , one by one in sequence, or copulate peptide fragments to the terminal amino acid, one by one in sequence, or copulate peptide fragments to the terminal amino acid. The amino groups that are not used in the coupling reaction are typically protected to prevent coupling in an incorrect location. If a solid phase synthesis is adopted, the C-terminal amino acid is bound to an insoluble carrier or support through its carboxyl group. The insoluble carrier is not particularly limited, as long as it has a binding capacity to a reactive carboxyl group. Examples of such insoluble carriers include halomethyl resins, such as chloromethyl resin or bromomethyl resin, hydroxymethyl resins, phenolic resins, ter-alkyloxycarbonyl hydrazide resins, and the like. An amino acid protected amino group is linked in sequence through the condensation of its activated carboxyl group and the reactive amino group of the previously formed peptide or chain, to synthesize the peptide step by step. After synthesizing the entire sequence, the peptide is cleaved from the insoluble carrier to produce the peptide. This solid phase approach is generally described by Merrifield et al. (1963), in J. Am. Chem. Soc. 85: 2149-2156. The prepared protein and fragments thereof can be isolated and purified from the reaction mixture by means of peptide separation, for example, by extraction, precipitation, electrophoresis and various forms of chromatography, and the like. The IL-170 proteins of this invention can be obtained in various degrees of purity, according to their intended use. The purification can be achieved by means of the desired use. Purification can be achieved through the use of the protein purification techniques described herein, or through the use of antibodies described herein in immunoabsorbent affinity chromatography. This immunoabsorbent affinity chromatography is carried out by first attaching the antibodies to a solid support, and then contacting the bound antibodies with solubilized lysates of cells from appropriate sources, lysates of other cells expressing the protein, or lysates or cell supernatants. that produce the IL-170 protein as a result of DNA techniques, see below.

V. Physical Variants This invention further encompasses proteins or peptides ng substantial amino acid sequence homology to the amino acid sequence of the IL-170 protein. Variants include allelic or species variants. The homology of amino acid sequences, or sequence identity, is determined by optimizing junctions of residues, if necessary, introducing spaces as required. This changes when conservative substitutions are considered as unions. Conservative substitutions typically include substitutions within the following groups: glycine, alanine, valine, isoleucine, leucine; aspartic acid, glutamic acid; asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. Homologous amino acid sequences are typically intended to include natural interspecies and allelic variations in each protein sequence respectively. Typical homologous peptides or proteins will from 25-100% homology (if spaces can be introduced), at 50-100% homology (if conservative substitutions are included) with the amino acid sequence of the IL-170 protein. The homology measurements will be at least about 35%, in general at least 40%, more generally at least 45%, often at least 50%, more often at least 55%, typically at least 60%, more typically at least 65%, usually at least 70%, more usually at least 75%, preferably at least 80%, and most preferably at least 80%, and in particularly preferred embodiments, at least 85% or more. See also Needleham et al. (1970) J. Mol. Biol. 48: 443-453; Sankoff and others (1983) Chapter one in Time Warps. Strinq Edits. and Macromolecules: The Theory and Practice of Sequence Comparison Addison-Wesley, Reading, MA; and software packages from IntelliGenetics, Mountain View, CA; and the University of Wisconsin Genetics Computer Group, Madison, Wl. The isolated DNA encoding an IL-170 protein can be easily modified by nucleotide substitutions, nucleotide deletions, nucleotide insertions and inversions of nucleotide stretches. These modifications produce new DNA sequences encoding these antigens, their derivatives, or proteins that similar antigenic, immunogenic or physiological activity. These modified sequences can be used to produce mutant antigens or to improve expression. Improved expression may involve gene amplification, increased transcription, increased translation, and other mechanisms. Said mutant IL-170 protein derivatives include predetermined or site-specific mutations of the respective protein or its fragments. "IL-170 mutant protein" encompasses a polypeptide that is otherwise within the definition of homology of the human IL-170 protein or murine IL-170, as set forth above, but ng an amino acid sequence that defers of the IL-170 protein as found in nature, either as a deletion, substitution, or insertion. In particular, "site-specific mutant protein IL-170" generally includes proteins ng significant homology to the corresponding protein ng the sequences of Tables 1-6, and which share several biological activities, eg, antigenic or immunogenic, with those sequences, and in preferred embodiments contain the majority of the described sequences. Similar concepts apply to different IL-170 proteins, particularly those found in several warm-blooded animals, e.g., mammals and birds. As stated above, it is emphasized that the descriptions in general are intended to encompass all IL-170 proteins, not limited to the mouse modality that is specifically discussed.

While site-specific mutation sites are predetermined, mutants do not need to be site-specific. The mutagenesis of IL-170 protein can be conducted by making amino acid insertions or deletions. The substitutions, deletions, insertions, or any combination, can be generated to arrive at a final construction. Inserts include amino- or carboxy-terminal fusions. Random mutagenesis can be conducted in an objective codon and the expressed mutants can then be screened to determine the desired activity. Methods for making substitution mutations at predetermined sites in DNA having a known sequence are well known in the art, for example, by mutagenesis of M13 primer or polymerase chain reaction (PCR) techniques. See also Sambrook et al. (1989) and Ausubel et al. (1987 and Supplements). Mutations in DNA should normally not place coding sequences outside of reading frames, and preferably will not create complementary regions that could hybridize to produce secondary mRNA structure such as loops or hairpins. The present invention also provides recombinant proteins, for example, heterologous fusion protein using segments of these proteins. A heterologous fusion protein is a fusion of proteins or segments that naturally are not normally fused in the same way. Thus, the fusion product of an immunoglobulin with an IL-170 polypeptide is a continuous protein molecule having sequences fused to a typical peptide linkage, typically made as a single translation product and exhibiting properties derived from each source peptide. . A similar concept is applied to heterologous nucleic acid sequences. In addition, new constructs can be made combining similar functional domains of other proteins. For example, antigen binding segments, or others, can be "exchanged" between different new fragments or fusion polypeptides. See, for example, Cunningham et al. (1989) Science 243: 1330-1336; and O'Dowd et al. (1988) J. Biol. Chem. 263: 15985-15992. Thus, novel chimeric polypeptides that exhibit novel combinations of specificities will result from functional binding of biologically relevant domains, and other functional domains. The phosphoramidite method described by Beaucage and Carruthers (1981) Tetra. Letts. 22: 1859-1862, will produce suitable synthetic DNA segments. A double strand fragment will often be obtained by either synthesizing the complementary strand and aligning or fixing the strands together under appropriate conditions, or by adding the complementary strand using DNA polymerase with an appropriate primer sequence, eg, PCR techniques.

SAW. Functional variants The blocking of the physiological response to IL-170 proteins can result from the inhibition of the antigen binding to its natural binding partner, for example, through competitive inhibition. Accordingly, the in vitro assays of the present invention will often utilize isolated protein, cell membranes expressing an IL-170 protein associated with recombinant membrane, soluble fragments comprising linker segments, or fragments attached to solid phase substrates. These assays will also allow the diagnostic determination of the effects of either mutations and modifications of linker segments, or mutations and protein modifications, eg, analogues. This invention further contemplates the use of competitive drug screening assays, for example, when neutralizing antibodies for antigen or binding partner fragments compete with an evaluating compound for binding to the protein. In this way, the antibodies can be used to detect the presence of any polypeptide that shares one or more antigenic binding sites of the protein, and can also be used to occupy binding sites on the protein that might otherwise interact with the protein. a link partner. Additionally, neutralizing antibodies against IL-170 protein and soluble fragments of the antigen that contain a high affinity receptor binding site can be used to inhibit antigen function in tissues, for example, tissues that undergo abnormal physiology. "Derivatives" of the IL-170 antigens include mutants of amino acid sequences, glycosylation variants and aggregated or covalent conjugates, with other chemical moieties. Covalent derivatives can be prepared by linking functionalities to groups found in amino acid side chains IL-170, or in the N- or C- terminus, by means that are well known in the art. These derivatives may include, without limitation, aliphatic esters or amides of the carboxyl terminus, or residues containing carboxyl side chains, O-acyl derivatives of residues containing hydroxyl groups and N-acyl derivatives of residues containing amino groups, or amido amino acids. terminals, for example, lysine or arginine. The acyl groups are selected from the group of alkyl portions including C3 to C18 normal alkyl, thus forming alkanoyl aroyl species. Covalent binding to carrier proteins can be important when the inumunogenic portions are haptens. In particular, glycosylation alterations are included, for example, made by modifying the glycosylation patterns of a polypeptide during its synthesis and processing, or in other processing steps. Particularly preferred means for effecting this are by exposing the polypeptide to glycosylating enzymes derived from cells that normally provide such processing, for example, mammalian glycosylation enzymes. Deglycosylation enzymes are also contemplated. In addition, versions of the same primary amino acid sequence having other minor modifications are included, including phosphorylated amino acid residues, for example, phosphotyrosine, phosphoserine, or phosphothreonine.

A major group of derivatives are covalent conjugates of the IL-170 protein or fragments thereof with other proteins or polypeptides. These derivatives can be synthesized in recombinant culture, such as N- or C-terminal fusions, or by the use of agents known in the art for their usefulness in cross-linking proteins through reactive side groups. Preferred antigen derivatization sites with crosslinking agents are free amino groups, carbohydrate moieties and cysteine residues. Fusion polypeptides between IL-170 proteins and other homologous or heterologous proteins are also provided. Homologous polypeptides can be fusions between different surface markers, producing, for example, a hybrid protein exhibiting receptor binding specificity. Likewise, heterologous fusions can be constructed, which would exhibit a combination of properties or activities of the derived proteins. Typical examples are fusions of an informant polypeptide, eg, luciferase, with a segment or domain of an antigen, eg, a receptor binding segment, such that the presence or location of the fused antigen can be readily determined. See, for example, Dull et al., U.S. Patent. No. 4,859,609. Other gene fusion partners include bacterial β-galactosidase, trpE, protein A, β-lactamase, alpha amylated, alcohol dehydrogenase and yeast alpha mating factor. See, for example, Godowski et al. (1998) Science 241: 812-816.

The phosphoramidite method described by Beaucage and Carruthers (1981) Tetra. Letts 22: 1859-1862, will produce suitable synthetic DNA fragments. A double-stranded fragment will often be obtained by either synthesizing the complementary strand and fixing the strands together under appropriate conditions, or by adding the complementary strand using DNA polymerase with an appropriate primer sequence. Said polypeptides may furthermore have amino acid residues which have been chemically modified by phosphorylation, sulfonation, biotinylation, or diction or removal of other portions, in particular those having molecular forms similar to phosphate groups. In some embodiments, the modifications will be reactive labeling useful, or serve as purification targets, e.g., affinity ligands. The fusion proteins will typically be made either by recombinant nucleic acid methods, or by synthetic polypeptide methods. Techniques for nucleic acid manipulation and expression are generally described, for example, in Sambrook et al. (1989) Molecular Cloninq: A Laboratory Manual (2nd Ed.), Vol. 1-3, Cold Spring Harbor Laboratory. Techniques for the synthesis of polypeptides are described, for example, in Merrifield (1963) J. Amer. Chem. Soc. 85: 2149-2156; Merrifield (1986) Sciences 232: 341-347; and Atherton et al. (1989) Solid Phase Peptide Synthesis: A Practice! Aproach, IRL Pres, Oxford.

This invention further contemplates the use of derivatives of IL-170 proteins that are not variations in the amino acid sequence or glycosylation. Said derivatives may involve covalent or aggregative association with chemical portions. These derivatives are generally found in the three classes: (1) salts, (2) covalent modifications of terminal and side chain residues and (3) absorption complexes, for example, with cell membranes. Said covalent or aggregative derivatives are useful as immunogens, as reagents in immunoassays, or in purification methods such as for affinity purification of antigens or other binding proteins. For example, an IL-170 antigen can be immobilized by covalent bonding to a solid support, such as Sepharose activated by cyanogen bromide, by methods that are well known in the art, or absorbed on polyolefin surfaces, with or without crosslinking. glutaraldehyde, for use in the assay or purification of anti-IL-170 protein antibodies or its receptor or other binding partner. IL-170 antigens can also be labeled with a detectable group, for example, radioiodinated by the chloramine T method, covalently linked to rare earth chelates, or conjugated to another fluorescent moiety for use in diagnostic assays. The purification of IL-17O protein can be effected by binding partners or immobilized antibodies. A solubilized IL-170 antigen or fragment of this invention can be used as an immunogen for the production of antiserum or antibodies specific for the protein or fragments thereof. The purified antigen can be used to screen monoclonal antibodies or binding fragments prepared by immunization with various forms of impure preparations containing the protein. In particular, the term "antibodies" also encompasses antigen binding fragments of natural antibodies. The purified IL-170 proteins can also be used as a reagent to detect any antibody generated in response to the presence of high levels of the protein or cellular fragments containing the antigen, which can be diagnosed as a specific abnormal or physiological condition. or of illness. Additionally to produce the antibodies of the present invention, as described immediately below. For example, this invention contemplates antibodies raised against amino acid sequences encoded by the nucleotide sequences shown in Tables 1-6., or fragments of proteins that contain them. In particular, this invention contemplates antibodies that have binding affinity or that are produced against specific fragments that are predicted to be found outside of the biphered lipid. The present invention contemplates the asylation of variants of additional closely related species. The Southern blot analysis established that similar genetic entities exist in other mammals, for example, rat and human. It is possible that IL-170 proteins are widely disseminated in variants of species, for example, rodents, lagomorphs, carnivores, artiodactyls, perissodactyls and primates.

The invention further provides means for isolating a group of related antigens that exhibit both differences and similarities in structure, expression and function. The elucidation of many of the physiological effects of the antigens will be greatly accelerated by the isolation and characterization of variants of different species. In particular, the present invention provides useful probes for identifying additional homologous genetic entities in different species. The isolated genes will allow the transformation of cells lacking the expression of a corresponding IL-170 protein, for example, either species or cell types lacking corresponding antigens and should exhibit negative background activity, the expression of transformed genes will allow the isolation of antigenically pure cell lines, with variants of unique or defined species. This approach will allow a more sensitive detection and discrimination of the physiological effects of IL-170 proteins. The subcellular fragments, for example, cytoplasts or fragments of membranes, can be insulator and used. Dissection of the critical structural elements that effect the various differentiating or physiological functions provided by the proteins is possible using standard techniques of modern molecular biology, particularly in the comparison of members of the related classes. See, for example, the homologous scanning mutagenesis technique described in Cunningham et al. (1989) Science 243: 1339-1336; and approaches used in O'Dowd et al. (1988) J. Biol-Chem 263: 15985-15992; and Lechleiter et al. (1990) EMBO J. 9: 4381-4390. In particular, domains or functional segments can be substituted between species variants to determine which structural characteristics are important both in binding partner affinity and in specificity, as well as signal transduction. A range of different variants will be used to track to determine molecules that exhibit combined properties of interaction with different species variants of link partners. The antigenicization of the antigen can occur under certain circumstances, and the interaction between intracellular components and "extracellular" segments of proteins involved in the interactions can occur. Specific segments of IL-170 protein interaction with other intracellular components can be identified by mutagenesis or direct biochemical means, for example, cross-linking or affinity methods. In addition, structural analysis will be applicable by crystallographic and other physical methods. Further investigation of the mechanism of biological function will include the study of associated components that can be isolated by affinity methods or by genetic means, for example mutant complementation analysis. Further study of expression and control of IL-170 protein will be pursued. The control elements associated with the antigens may exhibit specific tissue patterns, differential development, or other expression patterns. The ascending or descending genetic regions, for example, control elements, are of interest. Structural studies of the antigen will lead to the design of new variants, in particular analogs that exhibit agonist or antagonist properties, in particular analogs that exhibit agonist or antagonist properties on binding partners. This can be combined with screening methods described above, to isolate variants that exhibit spectra of desired activities. Expression in other cell types will often result in glycosylation differences in a particular antigen. Several species variants may exhibit different base functions to structural differences that are not amino acid sequences. Differential modifications may be responsible for the differential function, and the elucidation of the effects now becomes possible. Therefore, the present invention provides important reagents related to the interaction of binding partner-antigen. While the foregoing description has focused primarily on the human IL-170 protein and murine IL-170, those skilled in the art will immediately recognize that the invention encompasses other antigens, e.g., from mouse and other mammalian species or allelic variants, as well as variants thereof.

VIL Antibodies Antibodies can be produced for the various IL-170 proteins, including allelic species or variants, and fragments thereof, both in their natural forms and in their recombinant forms. Additionally, antibodies to IL-170 proteins can be produced either in their active forms or in their inactive forms. Anti-idiotypic antibodies are also contemplated. Antibodies including binding fragments and single chain versions, against predetermined fragments of the antigens, can be produced by immunization of animals with conjugates of the fragments with immunogenic proteins. The monoclonal antibodies are prepared from cells that secrete the desired antibody. These antibodies can be screened for binding to normal or defective IL-170 proteins, or they can be screened for agonist or antagonist activity, for example, mediated through a binding partner. These monoclonal antibodies will usually bind with at least one KD of about 1 mM, more usually at least about 300 μM, typically at least about 10 μM, more typically at least about 30 μM, preferably at least about 10 μM, and most preferably at least about 3 μM μM or more. An IL-170 polypeptide that specifically binds or that is specifically immunoreactive with an antibody, for example, such as a polyclonal antibody, generated against a defined immunogen, for example, such as an immunogen consisting of an amino acid sequence of SEQ ID NO. mature NO: 8 or fragments thereof or a polypeptide generated from the nucleic acid of SEQ ID NO: 7, is typically determined in an immunoassay. Included within the boundaries and limits of the present invention are those nucleic acid sequences as described herein, including functional variants, which encode polypeptides that selectively bind to polyclonal antibodies raised against the prototypical polypeptide IL-173, IL-174. , IL-176 or IL-177, as structurally and functionally described herein. The immunoassay typically uses a polyclonal antiserum that was produced, for example, for a protein of SEQ ID NO: 8. This antiserum is selected to have low cross-reactivity against other members of the appropriate IL-170 family, preferably the same species, and any such cross-reactivity is eliminated by inunoabsorption before use in the immunoassay. Appropriate selective serum preparations can be isolated and characterized. To produce antiserum for use in an immunoassay, the protein, for example, of SEQ ID NO: 8, is isolated as described herein. For example, recombinant protein can be produced in a mammalian cell line. An appropriate host, e.g., an inbred mouse strain such as Balb / c, is immunized with the protein of SEQ ID NO: 8 using a standard adjuvant, such as Freund's adjuvant, and a standard mouse immunization protocol (see Harlow and Lane). Alternatively, a synthetic peptide of substantially full length, derived from the sequences described herein, can be used as an immunogen. The polyclonal sera are collected and titrated against the immunogen protein in an immunoassay, for example, a solid phase immunoassay with the immunogen immobilized on a solid support. Polyclonal antisera with a titer of 104 or more are screened and evaluated for cross-reactivity against other members of the IL-170 family, eg, IL-171, IL-172, or IL-175, using an immunoassay of competitive link such as the one described in Lane, above, on pages 570-573. Preferably at least two members of the IL-170 family are used in this determination in conjunction with the objective. These members of the IL-170 family can be produced as recombinant proteins and isolated using standard molecular biology and protein chemistry techniques, as described herein. Thus, preparations of antibodies having desired selectivity or specificity for subgames of members of the IL-170 family can be identified or produced. Immunoassays in the competitive binding format can be used for cross-reactivity determinations. For example, the mature SEQ ID NO: 8 protein can be immobilized to a solid support. The proteins added to the assay compete with the binding of the antisera to the immobilized antigen. The ability of the above proteins to compete with the binding of the antisera to the immobilized protein is compared to the protein of SEQ ID NO: 8. The percent cross-reactivity for the above proteins is calculated, using standard calculations. These antisera with less than 10% cross-reactivity with each of the proteins listed above are selected and pooled. The cross-reactive antibodies are then removed from the pooled antisera by immunoabsorption with the proteins listed above. The immunosorbed and pooled antisera are then used in a competitive binding immunoassay as described above, to compare a second protein with the immunogen protein. To make this comparison, the two proteins are each assayed at a wide range of concentrations, and the amount of each protein necessary to inhibit 50% of the binding of the antisera to the immobilized protein is determined. If the amount of the second necessary protein is less than twice the amount of the protein of, for example, SEQ ID NO: 8 that is required, then the second protein is said to bind specifically to an antibody generated to the immunogen. The antibodies, including, antigen binding fragments, of this invention, may have significant diagnostic or therapeutic value. They can be potent antagonists that bind to a binding partner and inhibit the binding of antigen or inhibit the ability of an antigen to produce a biological response. They may also be useful as non-neutralizing antibodies, and may be coupled to toxins or radionuclides so that when the antibody binds to the antigen, a cell expressing it, for example, on its surface, dies. In addition, these antibodies can be conjugated with drugs or other therapeutic agents, either directly or indirectly by means of a linker, and can effect the identification of the drug. The antibodies of this invention may also be useful in diagnostic applications. As capture or non-neutralizing antibodies, they can be screened for their ability to bind antigens without inhibiting binding by a partner. As neutralizing antibodies, they can be useful in competitive binding assays. They will also be useful in the detection or quantification of IL-170 protein or its binding partners. See, for example, Chan (ed. 1987) Immunoassay: A Practical Guide Academic Press, Orlando, Fia.; Ngo (ed. 1988) Nonisotopic Immunoassavs Plenum Press, NY; and Price and Newman (eds. 1991) Principles and Practice of Immunoassay Stockton Press, NY. Fragments of antigens can be bound to other materials, in particular polypeptides, such as polypeptides fused or covalently linked to be used as immunogens. An antigen and its fragments can be fused or covalently linked to a variety of immunogens, such as limpet hemocyanin, bovine serum albumin, tetanus toxoid, etc. See Microbioloqy, Hoeber Medical Division, Harper and Row, 1969; Landsteiner (1962) Specificity of Serological Reactions. Dover Publications, New York, and Williams et al. (1967) Mehods in Immunology and Immunochemistry, Vol. 1, Academic Press, New York, for descriptions of methods for the preparation of polyclonal antisera. A typical method involves the hyperimmunization of an animal with an antigen. The animal's blood is then collected shortly after repeated immunizations, and gamma globulin is isolated. In some cases, it is convenient to prepare monoclonal antibodies from several mammalian hosts, such as mice, rodents, primates, humans, etc. The description of the techniques for the preparation of said monoclonal antibodies can be found, for example, in Stites et al. (Eds.) Basic and Clinical Immunology (4th ed.), Lange Medical Publications, Los Altos, CA, and references cited in the same; Harlow and Lane (1988) Antibodies: A Laboratorv Manual. CSH Press; Goding (1986) Monoclonal Antibodies: Principies and Practice (2nd ed.) Academic Press, New York; and in particular in Kohler and Milstein (1975) in Nature 256: 495-497, which discusses a method for the generation of monoclonal antibodies. Briefly summarized, this method involves injecting an animal with an immunogen. The animal is then sacrificed and the cells of the arm are taken, which are then fused with myeloma cells. The result is a hybrid cell or "hybridoma", which is capable of reproducing in vitro. The population of hybridomas is then screened to isolate individual clones, each of which secretes a single species of antibody to the immunogen. In this way, the individual antibody species obtained are the products of simple B cells cloned and immortalized, from the immune animal generated in response to a specific site recognized in the immunogenic substance.

Other suitable techniques involve the in vitro exposure of lymphocytes to the antigenic polypeptides, or alternatively, the selection of antibody libraries in phage or similar vectors. See, Huse et al. (1989) "Generation of a Large Combinatorial Library of the Immuoglobulin Repertoire in Phage Lambda", Science 246: 1275-1281; and Ward et al. (1989) Nature 341: 544-546. The polypeptides and antibodies of the present invention can be used with or without modification, including chimeric or humanized antibodies. Frequently, polypeptides and antibodies will be labeled by binding, either covalently or non-covalently, a substance that provides a detectable signal. A wide variety of labels and conjugation techniques are known, and are reported extensively in both the scientific and patent literature. Suitable labels include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent moieties, chemiluminescent portions, magnetic particles, and the like. The patents that teach the use of said labels include the Patents of E.U.A. Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241. In addition, recombinant immunoglobulins can be produced; see Cabilly, Patent of E.U.A. No. 4,816,567. The antibodies of this invention can also be used for affinity chromatography in the isolation of the protein. Columns can be prepared where the antibodies bind to a solid support, for example, particles, such as agarose, Sephadex, or the like, where a cell lysate can be passed through the column, the column washed, followed by increasing concentrations. of a soft denaturant, which will release purified IL-170 protein. The antibodies can also be used to screen expression libraries for particular expression products. Usually the antibodies used in said procedure will be labeled with a portion allowing the easy detection of the presence of antigen by antibody binding. The antibodies raised against each IL-170 protein will be useful to produce anti-idiotypic antibodies. These will be useful in the detection or diagnosis of various immunological conditions related to the expression of the respective antigens.

VIII. Uses The present invention provides reagents that will find use in diagnostic applications as described elsewhere herein, for example, in the general description for physiological or developmental abnormalities, or thereafter, in the description of diagnostic equipment. This invention also provides reagents with significant therapeutic value. The IL-170 protein (natural or recombinant), fragments thereof, and antibodies thereto, together with compounds identified to have binding affinity to IL-170 protein should be useful in the treatment of conditions associated with development or physiology. abnormal, including abnormal proliferation, for example, cancerous conditions, or degenerative conditions. Abnormal proliferation, regeneration, degeneration and atrophy can be modulated by appropriate therapeutic treatment, using the compositions provided herein. For example, a disease or disorder associated with abnormal expression or abnormal signaling by an IL-170 antigen should be a possible target for a protein agonist or antagonist. Other conditions of abnormal development are known in the cell types that show to possess IL-170 antigen mRNA by Northern blot analysis. See Berkow (ed.) The Merck Manual of Diagnosis and Therapy. Merck & Co., Rahway, N. J .; and Thorn and others Harrison's Principies of Intemal Medicine. McGraw-Hill, N. Y. These problems may be susceptible to prevention or treatment using the compositions provided herein. Recombinant antibodies that bind to IL-170 can be purified and then administered to a patient. These reagents can be combined for therapeutic use with inert ingredients or additional active ingredients, for example, in conventional pharmaceutically acceptable carriers or diluents, for example, immunogenic adjuvants, together with physiologically harmless stabilizers and excipients. These combinations can be sterile filtered and placed in dosage forms, such as by lyophilization, in dosing containers, or storage in stabilized aqueous preparations. This invention also contemplates the use of antibodies or binding fragments thereof, including forms that are not complement binders. A screening can be performed using IL-170 for binding partners or compounds that have binding affinity to IL-170 antigen, including isolation of associated components. Subsequent biological assays can then be used to determine whether the compound has intrinsic biological activity, and is therefore an agonist or antagonist in that it blocks an activity of the antigen. This invention also contemplates the therapeutic use of antibodies to the IL-170 protein as antagonists. This approach should be particularly useful with other variants of IL-170 protein species. The amounts of reagents necessary for effective therapy will depend on many different factors, including means of administration, target sites, physiological status of the patient, and other medications administered. Thus, treatment dosages should be titrated to optimize safety and efficacy. Typically, dosages used in vitro can provide useful guidance in amounts useful for in situ administration of these reagents. The evaluation of animals of effective doses for the treatment of particular disorders will provide an additional predictive indication of human dosage. Several considerations are described, for example, in Gilman et al. (Eds. 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 8th Ed., Pergamon Press; and Remington's Pharmaceuticals Sciences. 17th Ed. (1990), Mack Publishing Co., Easton, Penn. Methods for administration are discussed therein and then, for example, for oral, intravenous, intraperitoneal, or intramuscular administration, transdermal diffusion and others. See also Langer (1990) Science 249: 1527-1533. The pharmaceutically acceptable carriers will include water, saline, pH regulators and other compounds that are described, for example, in the Merck Index, Merck & Co., Rahway, New Jersey. It would be commonly expected that the dosing ranges be in amounts less than concentrations of 1 mM, typically lower than concentrations of about 10 μM, usually less than about 100 nM, preferably less than about 10 pM (picomolar), and more preferably, less than approximately 1fM (femtomolar), with an appropriate carrier. Slow release formulations, or a slow release device, will often be used for continuous administration. The IL-170 protein, fragments thereof and antibodies thereto or their fragments, antagonists and agonists, can be administered directly to the host to be treated, or according to the size of the compounds, it may be convenient to conjugate them with carrier proteins of such as ovalbumin serum albumin before its administration. The therapeutic formulations can be administered in any conventional dosage formulation. While it is possible for the active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation. The formulations typically comprise at least one active ingredient, as defined above, together with one or more acceptable carriers thereof. Each carrier should be both pharmaceutically and physiologically acceptable, in the sense of being compatible with the other ingredients, and not harmful to the patient. The formulations include suitable for oral, rectal, nasa, or parenteral administration (including subcutaneous, intramuscular, intravenous and intradermal). The formulations may conveniently be presented in unit dosage form, and may be prepared by any method well known in the art of pharmacy. See, for example, Gilman et al. (Eds. 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics. 8th Ed., Pergamon Press, Parr town, NY; Remington's Pharmaceuticals Sciences. 17th Ed. (1990), Mack Publishing Co., Easton, Penn .; Avis et al. (Eds. 1993) Pharmaceutical Dosage Forms: Parenteral Medications 2nd. Ed., Dekker, NY; Lieberman et al. (Eds. 1990) Pharmaceutical Dosaqe Forms: Tablets 2nd. Ed., Dekker, NY; and Lieberman et al. (eds. 1990) Pharmaceutical Dosage Forms: Disperse Systems Dekker, NY. The therapy of this invention can be combined or used in association with other therapeutic reagents, including cytokine. Both the natural and recombinant formulas of the IL-170 proteins of this invention are particularly useful in kits and assay methods that are capable of screening compounds for the determination of protein binding activity. Several methods of automated testing have been developed in recent years, in order to allow the tracking of tens of thousands of compounds in a short period. See, for example, Fodor et al. (1991) Science 251: 767-773, which describes means for evaluating binding affinity by a plurality of defined polymers synthesized on a solid substrate. The development of suitable assays can be greatly facilitated by the availability of large amounts of soluble, purified IL-170 protein, as provided by this invention. This invention is particularly useful for the screening of compounds using recombinant antigen in any of a variety of drug screening techniques. The advantages of using a recombinant protein in screening for specific ligands include, (a) improved renewable source of the antigen from a specific source, (b) potentially greater number of antigen molecules per cell, providing better signal to noise ratio in the essays; and (c) specificity of variety of species (theoretically providing greater biological and disease specificity). The purified protein can be evaluated in numerous assays, typically in in vitro assays, which evaluate biologically important responses. See for example, Coligan Current Protocols in Immunoloqy; Hood and other Immunology Benjamin / Cummings; Paul (ed.) Fundamental Immunology; and Methods in Enzymology Academic Press. A method of drug screening utilizes eukaryotic or prokaryotic host cells that are stably transformed with recombinant DNA molecules that express the IL-170 antigens. Cells expressing an antigen isolated from other functionally equivalent antigens can be isolated. Said cells, either in a viable or fixed form, can be used for standard protein-protein binding assays. See also, Parce et al., (1989) Science 246: 243-247, and Owicki et al. (1990) Proc. Nat'l Acad. Sci. USA 87: 4007-4011, which describes sensitive methods for detecting cellular responses. Competitive assays are particularly useful, where the cells (IL-170 protein source) are contacted and incubated with an antibody or labeled binding partner having known binding affinity to the ligand, such as 125I antibody, and a sample of evaluation whose binding affinity to the link composition is being measured. The free labeled and bound binding compositions are then separated to evaluate the degree of antigen binding. The amount of bound evaluation compound is inversely proportional to the amount of receptor bond labeled to the known source. Any of numerous techniques can be used to separate antigen bound from the free, to evaluate the degree of binding. This separation step could typically involve a procedure such as adhesion to filters, followed by washing, adhesion to plastic followed by washing, or centrifugation of the cell membranes. In adon, viable cells could be used to screen for the effects of the drugs on IL-170 protein-mediated functions, eg, secondary messenger levels, i.e., Ca ++, cell proliferation, inositol phosphate binding changes; and others.

Some detection methods allow the elimination of a separation step, for example, a sensitive proximity detection system. Calcium sensitive dyes will be useful for the detection of Ca ++ levels, with a fluorescence cell selection apparatus, or a fluorimeter. Other method uses transformed prokary or eukary host cell membranes as the source of the IL-170 protein. These cells are stably transformed with DNA vectors that direct the expression of an IL-170 protein associated with a membrane, for example, a designed membrane-bound form. Essentially, the membranes would be prepared from the cells, and would be used in any receptor / ligand binding assay, such as the competitive assay discussed above. Yet another approach is the use of purified, solubilized or purified solubilized IL-170 protein from transformed eukary or prokary host cells. This allows a "molecular" binding assay, with the advantages of increased specificity, the automation capability, and high drug evaluation performance. Another technique for drug screening involves an approach that provides high throughput screening of compounds that have adequate binding affinity to IL-170, and is described in detail in Geysen, European Patent Application 84/03564, published on 13 September 1984. First, large numbers of different small peptide evaluation compounds are synthesized on a solid substrate, for example, plastic pins or some other appropriate surface; see Fodor et al., (1991). All the pins are then reacted with solubilized, purified, solubilized or solubilized IL-170 binding composition, and washed. The next step involves detecting linked link composition. A rational drug design can also be based on structural studies of the molecular forms of the IL-170 protein and other effectors or the like. The effectors may be other proteins that mediate other functions in response to antigen binding, or other proteins that normally interact with the antigen. A means of determining which sites interact with other specific proteins is a determination of physical structure, for example, X-ray crystallography or 2-dimensional NMR techniques. This will provide guidance as to which amino acid residues form molecular contact regions. For a detailed description of the structural determination of proteins, see, for example, Blundell and Johnson (1976) Protein Crystallography. Academic Press, New York. Plates can be coated directly with the IL-170 protein, for use in the aforementioned drug screening techniques. However, non-neutralizing antibodies to these ligands can be used as capture antibodies to immobilize the respective ligand on the solid phase.

IX. Equipment This invention further contemplates the use of IL-170 proteins, fragments thereof, peptides and their fusion products, in a variety of diagnostic equipment and methods, for the detection of the presence of a binding composition. Typically, the kit will have a compartment containing either a defined IL-170 peptide, or segment; genetic, or a reagent that recognizes one or the other, for example, antibodies or antigen fragments. A kit for determining the binding affinity of an evaluation compound to an IL-170 protein would typically comprise an evaluation compound; a labeled compound, for example, an antibody having known binding affinity for the antigen; a source of protein IL-170 (natural or recombinant); and a means for separating labeled free compound from binding, such as a solid phase to immobilize the antigen. Once the compounds are screened, those that have adequate binding affinity to the antigen can be evaluated in appropriate biological assays, as they are well known in the art, to determine if they exhibit biological activities similar to the natural antigen. The availability of recombinant IL-170 protein polypeptides also provides well-defined standards for calibrating such assays. A preferred equipment for determining the concentration of, for example, an IL-170 protein in a sample would typically comprise a labeled compound, eg, antibody, which has known binding affinity for the antigen, a source of antigen (natural or recombinant) and a means for separating the free labeling compound from the linkage, eg, a solid phase to immobilize the IL-170 protein. Normally, compartments containing the reagents, and instructions will be provided. A method for determining the concentration of IL-170 protein in a sample would typically comprise the steps of: (1) preparing membranes from a sample composed of a membrane-bound protein source IL-170; (2) wash the membranes and suspend them in a buffer; (3) solubilizing the antigen, incubating the membranes in a culture medium to which a suitable detergent has been added; (4) adjust the detergent concentration of the solubilized antigen; (5) contacting and incubating said dilution with radio-labeled antibody to form complexes; (6) recovering the complexes, such as by filtration through treated polyethyleneimine filters; and (7) measure the radioactivity of the recovered complexes. Antibodies, including antigen binding fragments, specific for IL-170 protein or fragments, are useful in diagnostic applications to detect the presence of high levels of IL-170 protein and / or its fragments. Such diagnostic assays may employ lysates, living cells, fixed cells, immunofluorescence, cell cultures, body fluids and may also involve the detection of protein related antigens in serum, or the like. Diagnostic assays can be homogeneous (without a separation step between the free reagent and protein-protein complex) or heterogeneous (with a separation step). Several commercial assays exist, such as radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), enzyme immunoassay (EIA), multiplied enzyme immunoassay (EMIT) technique, labeled substrate fluorescent immunoassay (SLFIA) and the like. For example, unlabeled antibodies can be used using a second antibody that is labeled and recognizes the antibody for an IL-170 protein or for a particular fragment thereof. Similar trials in the literature have also been discussed extensively in the literature. See, for example, Harlow and Lane (1988) Antibodies: A Laboratory Manual. CSH Anti-idiotypic antibodies may have similar use to diagnose the presence of antibodies against an IL-170 protein, such as the diagnosis of several abnormal conditions. For example, overproduction of IL-170 protein can result in the production of various immunological reactions that can be diagnostic of abnormal physiological states, in particular in conditions of proliferating cells such as cancer or abnormal differentiation. Frequently, the reagents for the diagnostic tests are supplied in equipment, in order to optimize the sensitivity of the assay. For the present invention, according to the nature of the assay, protocol and label, either labeled or unlabeled antibody, or labeled IL-170 protein is provided. This is usually found in conjunction with other additives, such as buffers, stabilizers, materials necessary for signal production, such as substrates for enzymes, and the like. Preferably, the equipment will also contain instructions for the proper use and disposal of the contents after use. Typically, the equipment has compartments for each useful reagent. Conveniently, the reagents are provided as a dry lyophilized powder, where the reagents can be reconstituted in an aqueous medium that provides the appropriate concentrations of reagents to perform the assay. Any of the aforementioned constituents of the drug screening, and of the diagnostic assays, can be used without modification, or they can be modified in a variety of ways. For example, labeling can be achieved by covalently or non-covalently attaching a portion that directly or indirectly provides a detectable signal. In any of these assays, in antigen, evaluation compound, IL-170 protein, or antibodies thereto, they can be labeled either directly or indirectly. The possibilities for direct labeling include label groups: radiolabeled such as 125l, enzymes (US Patent No. 3,645,090), such as peroxidase and alkaline phosphatase, and fluorescent labels (US Patent No. 3,940,475) capable of monitoring the intensity change of fluorescence, wavelength change, or fluorescence polarization. The possibilities for indirect labeling include biotinylation of a constituent followed by the coupled avidin linkage of the groups of labels mentioned above.

In addition, there are numerous methods for separating the bound antigen from the free, or alternatively, the binding compound bound from the free. The IL-170 protein can be immobilized on several matrices, followed by washing. Suitable matrices include plastic, such as an ELISA plate, filters and beads. Methods for immobilization of the IL-170 protein to a matrix include, without limitation, direct adhesion to plastic, use of a capture antibody, chemical coupling and biotin-avidin. The last step in this approach involves the precipitation of protein-protein complex by any of several methods, including those using, for example, an organic solvent such as polyethylene glycol, or a salt such as ammonium sulfate. Other suitable separation techniques include, without limitation, the fluorescein antibody magnetizable particle method described in Rattle et al., (1984) Clin. Chem. 30: 1457-1461, and magnetic particle separation of double antibody as described in U.S. Patent No. 4,659,678. Methods for binding proteins or their fragments to the various labels have been reported extensively in the literature, and do not need detailed description in the present. Many of the techniques involve the use of activated carboxyl groups, either through the use of carbodiimide or active esters to form peptide bonds, the formation of thioethers by reaction of a mercapto group with an activated halogen, such as chloroacetyl, or a activated olefin such as maleimide, for binding, or the like. The fusion proteins will also find use in these applications. Another diagnostic aspect of this invention involves the use of oligonucleotide or polynucleotide sequences taken from the sequence of an IL-170 protein. These sequences can be used as probes to detect antigen message levels in samples from patients suspected of having an abnormal condition, eg, cancer or developmental problem. The preparation of both DNA and RNA nucleotide sequences, the labeling of the sequences, and the preferred size of the sequences, has received ample description and discussion in the literature. Normally, an oligonucleotide probe should have at least about 14 nucleotides, usually at least about 18 nucleotides, and the polynucleotide probes can be up to several kilobases. Several labels can be used, most commonly radionuclides, in particular 32P. However, other techniques can also be employed, such as the use of modified biotin nucleotides for introduction into a polynucleotide. Biotin then serves as the site for binding to avidin or antibodies, which can be labeled with a wide variety of labels, such as radionuclides, fluorescers, enzymes, or the like. Alternatively, antibodies can be used that can recognize specific pairs, including DNA pairs, RNA pairs, hybrid DNA-RNA pairs, or DNA-protein pairs. In turn, the antibodies can be labeled and the assay can be carried out, where the duple is bound to a surface, so that with the formation of the duple on the surface, the presence of antibody bound to the dupl can be detected. The use of probes for the new antisense RNA can be carried out in any conventional technique, such as nucleic acid hybridization, plus and minus screening, recombinatory probing, hybrid released translation (HRT) and hybrid stopped translation (HART). This also includes amplification techniques, such as polymerase chain reaction (PCR). Another approach uses, for example, antisense nucleic acid, including the introduction of double-stranded RNA (dsRNA), to genetically interfere with genetic function as described, for example, in Misquitta et al., (1999) Proc. Nat'l. Acad. Sci. E.U.A. 96: 1451-1456, and / or ribozymes to block the translation of a specific IL-170 mRNA. The use of antisense methods to inhibit in vitro translation of genes is well known in the art. Marcus-Sakura (1988) Anal. Biochem. 172: 289; Akhtar (ed. 1995) Deliverv Strategies for Antisense Oligonucleotide Therapeutics CRC Press, Inc. In addition, diagnostic kits are also evaluated to determine the qualitative or quantitative presence of other markers. The diagnosis or prognosis may depend on the combination of multiple indications used as markers. Thus, teams can evaluate for combinations of markers. See, for example, Viallet et al., (1989) Proqress in Growth Factor Res. 1: 89-97.

The broad scope of this invention is best understood with reference to the following examples, which are not intended to limit the invention to specific embodiments.

EXAMPLES I. General methods Some of the standard methods are described, or are referred to, for example, in Maniatis et al. (1982) Molecular Cloning. A Laboratory Manuals. Cold Spring Harbor Laboratory, Cold Spring Harbor Press; Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual. (2nd ed.). vol. 1-3, CSH Press, NY; Ausubel et al., Bioloqy. Greene Publishíng Associates, Brooklyn, NY; or Ausubel et al., (1987 and Supplements) Current Protocols in Molecular Bioloqy. Greene / Wiley, New York; Innis, et al., (Eds. 1990) PCR Protocols: A Guide to Methods and Applications Academic Press, N. Y .; and Kohler, et al., (1995) Quantitation of mRNA by Polvmerase Chain Reaction Springer-Verlag, Berlin. Methods for protein purification include methods such as ammonium sulfate precipitation, column chromatography, electrophoresis, centrifugation, crystallization and others. See, for example, Ausubel, et al., (1987 and periodic supplements); Deutscher (1990) "Guide to Protein Purification" in Methods in Enzymology, vol. 182, and other volumes in this series; and manufacturer's literature on the use of protein purification products, for example Pharmacia, Piscataway, N.J., or Bio-Rad, Richmond, CA. The combination with recombinant techniques allows fusion to appropriate segments, for example, to a FLAG sequence or an equivalent that can be fused by means of a protease removal sequence. See, for example, Hochuli (1989) Chemische Industrie 12: 69-70; Hochuli (1990) "Purification of Recombinant Proteins with Metal Chelate Absorbent" in Setlow (ed.) Genetic Enqineerinq. Principie and Methods 12: 87-98, Plenum Press, N.Y .; and Crowe et al., (1992) QIAexpress: The Hiqh Level Expression & Protein Purification System QUIAGEN, Inc., Chatsworth, CA. In addition, a similar patent application directed to the cytokines IL-171 and IL-175, Proxy case number DX0918P, filed on the same date as this one is incorporated herein by reference. Standard immunological techniques are described, for example, in Hertzenberg et al., (Eds. 1996) Weir's Handbook of Experimental Immunology vol. 1-4, Blackwell Science; Coligan (1991) Current Protocols in Immunoloqy and Wiley / Greene, NY; and Methods in Enzvmology vol. 70, 73, 74, 84, 92, 93, 108, 116, 121, 132, 150, 162 and 163. Cytokine assays are described, for example, in Thomson (ed., 1998) The Cytokine Handbook (3rd ed. ). Academic Press. San Diego; Mire-Sluis and Thorpe (1998) Cvtokines Academic Press, San Diego; Metcalf and Nicola (1995) The Hematopoietic Colony Stimulatinq Factors Cambridge University Press; and Aggarwal and Gutterman (1991) Human Cvtokines Blackwell Pub.

Assays for vascular biological activities are well known in the art. They will cover angiogenic and angiostatic activities in tumor, or other tissues, for example, arterial smooth muscle proliferation (see, for example, Koyoma et al., (1996) Cell 87: 1069-1078), adhesion of monocytes to the vascular epithelium (see McEvoy et al., (1997) J. Exp. Med. 185: 2069-2077), etc. See also Ross (1993) Nature 362: 801-809; Rekhter and Gordon (1995) Am. J. Pathol. 147: 668-677; Thyberg et al., (1990) Atherosclerosis 10: 966-990; and Gumbiner (1996) CeN 84: 345-357. Assays for determination of the biological activities of neural cells are described, for example, in Wouteriood (ed. 1995) Neuroscience Protocols Modules 10, Elsevier; Methods in Neurosciences Academic Press; and Neuromethods Humana Press, Totowa, NJ. The methodology of development systems is described, for example, in Meisami (ed.) Handbook of Human Growth and Developmental Biology CRC Press; and Chrispeeis (ed.) Molecular Techniques and Approaches in Developmental Biology Interscience. The analysis of computer sequences is carried out, for example, using available software programs, including those from the GCG (U. Wisconsin) and GenBank sources. In addition, public sequence databases were used, for example, from GenBank and others. Many techniques applicable to IL-170 can be applied to these new entities, as described, for example, in USSN, each of which is incorporated herein by reference for all purposes.

FACS analyzes are described in Melamed et al., (1990) Flow Cvtometry and Sorting Wiley-Liss, Inc., New York, NY; Shapiro (1988) Practical Flow Cytometry Liss, New York, NY; and Robinson et al., (1993) Handbook of Flow Cytometry Methods Wiley-Liss, New York, NY.

II. Isolation of a DNA clone encoding IL-170 protein Isolation of murine CTLA-8 is described in Rouvier et al., (1993) J. Immunol. 150: 5445-5456. Similar methods are available for isolation of counterpart species of IL-173, IL-174, IL-176 and IL-177, together with IL-171, IL-172 and IL-175.

Source of IL-170 messages Several cell lines are scanned using an appropriate probe for high-level message expression. Appropriate cell lines are selected based on the expression levels of the appropriate IL-170 message.

Isolation of an IL-170 coding clone Standard PCR techniques are used to amplify an IL-170 genetic sequence from a genomic or cDNA library, or from mRNA. A cDNA or genomic library is obtained and screened with a suitable synthetic or cDNA probe. PCR primers can be prepared. Appropriate primers are selected, for example, from the sequences provided, and a full-length clone is isolated. Various combinations of primers, of various lengths and possibly with differences in sequence, can be prepared. The full-length clone can be used as a hybridization probe to screen for other homologous genes using severe or less severe hybridization conditions. In another method, oligonucleotides are used to screen a library. In combination with polymerase chain reaction (PCR) techniques, synthetic oligonucleotides are used in appropriate orientations, as primers to select the correct clones from a library.

III. Biozymatic Characterization of IL-170 Proteins An IL-170 protein is expressed in heterologous cells, eg, the wild type or a recombinant form that exhibits the FLAG peptide at the carboxy terminus. See, for example, Crowe et al., (1992) QIAexpress: The High Level Expression and Protein Purification System QIAGEN, Inc. Chatsworth, CA; and Hopp et al., (1988) Bio / Technology 6: 1204-1210. These two forms are introduced into expression vectors, for example, pME18S, or pEE12, and subsequently transfected into appropriate cells, eg, COS-7 or NSO cells, respectively. Electroporated cells are cultured, for example, for 48 hours in RPMI medium supplemented with 10% Fetal Calf Serum. The cells are then incubated with 35S-Met and 35S-Cys, to label the cellular proteins. Comparison of the proteins under reducing conditions in SDS-PAGE should show that cells transfected with full-length clones should secrete a polypeptide of the appropriate size, for example about 15,000 daltons. Treatment with endoglycosidases will show if there are N-glycosylated forms.

IV. Large Scale Production Purification of IL-170 For biological assays, mammalian IL-170 is produced in large quantities, for example, with transfected COS-7 cells grown in RPMI medium supplemented with Nutridoma HU 1% (Boehringer Mannheim, Mannheim, Germany) and subsequently purified . The purification can use affinity chromatography using antibodies, or protein purification techniques, for example, using antibodies to determine the separation properties. To produce large amounts of natural proteins, stable transformants of NSO cells can be prepared according to the methodology developed by Celltech (Slough, Berkshire, UK: International patent applications WO 86/05807, WO 87/04462, WO 89/01036 and WO 89/10404). Typically, 1 liter of supernatant containing human IL-173 or IL-173-FLAG is passed, for example, on a 60 ml column of grafted Zn ++ ions to a Fast Flow Sepharose Chelating Matrix.

(Pharmacia, Upsalla, Sweden). After washing with 10 volumes of binding buffer (His-Bind Buffer kit, Novagen, Madison, Wl), the proteins retained by the metal ions are eluted with a gradient of 20-100 mM Imidazole. The content of human IL-173-FLAG in the eluted fractions is determined by dot blots using the monoclonal anti-FLAG antibody M2 (Eastman Kodak, New Haven, CT), while the content of human IL-173 is evaluated, by example, by silver staining of non-reducing SDS-PAGE. Fractions containing IL-170 are then pooled and dialysed against PBS, and either used in biological assays or additionally purified, for example, by anion exchange HPLC or on a DEAE column. A third step of gel filtration chromatography can be performed on a SUPERDEX G-75 HRD30 column (Pharmacia Uppsala, Sweden). The purification can be evaluated, for example, by SDS-PAGE with silver staining.

V. Preparation of antibodies against IL-173 Inbred Balb / c mice are immunized intraperitoneally, for example, with 1 ml of purified human IL-173-FLAG emulsified in complete Freund's adjuvant on day 0, and in incomplete Freund's adjuvant on days 15 and 22. The mice are stimulated with 0.5 ml of purified human IL-173 administered intravenously. The polyclonal antiserum is collected. The serum can be purified for antibodies. The antibodies can be further processed, for example, to Fab, Fab2, Fv or similar fragments. Hybridomas are created using, for example, non-secreting myeloma cells line SP2 / 0-Ag8 and polyethylene glycol 1000 (Sigma, St.

Louis, MO) and the fusion agent. Hybridoma cells are placed in a 96 container Falcon tissue culture dish (Becton Dickinson, NJ) and fed with F12 DMEM (Gibco, Gaithersburg, MD) supplemented with 80 μm / ml gentamicin, 2 mM glutamine, horse 10% (Gibco, Gaithersburg, MD), ADCM 1% (CRTS, Lyon, France) 10"5 M azaserin (Sigma, St. Louis, MO) and 5 x 10 ~ 5 M hypoxanthine. Hybridoma supernatants are screened for determination of production of antibodies against human IL-173, by immunocytochemistry (ICC) using COS-7 cells transfected with human IL-173 fixed in acetone, and by ELISA using purified human IL-173-FLAG from COS-7 supernatants as a coating antigen Aliquots of positive cell clones are expanded for 6 days, and cryopreserved, as well as propagated in ascites of Balb / c mice treated with pristane (2, 6, 10, 14-teramethylpentadecane, Sigma, St. Louis, MO) that had received with intraperitoneal injection of pristane 15 d Typically, approximately 105 hybridoma cells are provided in 1 ml of PBS, intraperitoneally, and 10 days later, ascites is collected from each mouse. After centrifugation of the ascites, the antibody fraction is isolated by precipitation of ammonium sulfate and anion exchange chromatography on a Zephyr-D silicon column (IBF Sepracor) equilibrated with 20 mM Tris pH 8.0. The proteins are eluted with a gradient of NaCl (ranging from 0 to 1 M NaCl). 2 ml fractions are collected and evaluated by ELISA to detect the presence of anti-IL-173 antibody. Fractions containing specific anti-IL-173 activity are pooled, dialyzed and frozen. They can be labeled with peroxidase, aliquots of purified monoclonal antibodies. Antibody preparations, polyclonal or monoclonal, can be cross-absorbed, reduced, or combined to create reagents exhibiting desired combinations of selectivity and specificities. Specific defined antigens can be immobilized to a solid matrix and used to selectively decrease or select desired binding capabilities.

SAW. Quantitation of human IL-173 Among the antibodies specific for IL-173, appropriate clonal isolates are selected to quantitate the levels of human IL-173 using a sandwich assay. The purified antibodies are diluted, for example, to 2 μg / ml in coating buffer (carbonate buffer, pH 9.6 15 mM Na2CO3, 35 mM NaHCO3). With this diluted solution, the containers of a 96-well ELISA plate (Immunoplate Maxisorp F96 certified, NUNC, Denmark) are coated overnight at room temperature. The plates are then washed manually, for example with washing buffer consisting of Saline with Phosphate Buffer and 0.05% Tween 20 (Technicon Diagnositics, USA). 110 μl of purified human CTLA-8 diluted in TBS-B-T buffer [20 mM Tris, 150 mM NaCl, 1% BSA (Sigma, St. Louis, MO) and 0.05% Tween 20] are added to each vessel. After 3 hours of incubation at 37 ° C, the plates are washed once. Add 100 μl of labeled peroxidase Ab diluted to 5 μg / ml in TBS-B-T buffer, to each container, and incubate for 2 hours at 37 ° C. The containers are then washed three times in washing buffer. Add 100 μl of peroxidase substrate, 2,2 'Azino-bis (3-ethylbenzthiazoin-6-sulfonic acid) (ABTS), diluted at 1 mg / ml in citrate / phosphate buffer, to each container, and read the reaction colorimetric at 405 nm.

Vile. Gene distribution IL-170 Human IL-173 was identified from sequence derived from a cDNA library of an epileptic cerebral frontal cortex. Rat IL-173 derived from a cDNA library of cochlea, brain, cerebellum, eye, lung and kidney. Again, the genes appear to be quite rare, suggesting that the expression distributions would be highly restricted. The mouse IL-174 of sequence derived from a cDNA library derived from a mouse embryo was identified. The gene seems to be quite rare, suggesting that the distribution of expression would be highly restricted. Human IL-171 was identified from a sequence derived from an apoptotic T cell. The gene seems to be quite rare, suggesting that the distribution of expression would be highly restricted. Human IL-172 was identified from sequences derived from human fetal heart, liver and spleen, thymus, thymic tumor and total fetus. Mouse was derived from sequences derived from mouse, embryo, mammary gland and assembled organs. Both genes appear to be quite rare, suggesting that their expression distribution would be highly restricted. The human IL-175 of a sequence derived from a T cell activated by thiouridine 12 h was identified. The gene seems to be quite rare, suggesting that the distribution of expression would be highly restricted.

VIII. Gene chromosome mapping IL-170 An isolated cDNA encoding the appropriate IL-170 gene is used. Chromosome mapping is a standard technique. See, for example, BIOS Laboratories (New Haven, CT) and methods for the use of a hybrid panel of mouse somatic cells with PCR. The human IL-173 gene maps to human chromosome 13q11.

IX. Isolation of IL-170 Homologs A binding composition, eg, antibody, is used for the screening of an expression library made of a cell line expressing an IL-170 protein. Standard staining techniques are used to detect or classify antigen expressed on the surface or intracellular, or transformed cells that express on surface are tracked by panning. The screening of intracellular expression is carried out by various staining or immunofluorescence procedures. See also McMahan et al., (1991) EMBO J. 10: 2821-2832.

Similar methods are applicable to isolate either allelic variants or species. Species variants are isolated using cross-species hybridization techniques based on a full-length isolate or fragment of a species such as a probe, or appropriate species.

X. Isolation of receptors for IL-170 Methods for the screening of an expression library made from a cell line expressing potential IL-170 receptors are available. A labeled IL-170 ligand is produced, as described above. Standard staining techniques are used to detect or classify surface expressed receptor, or transformed cells that express on surface are tracked by panning. See also McMahan et al., (1991) EMBO J. 10: 2821-2832. For example, on day 0, pre-coat 2-chamber permanox slides, with 1 ml per fibronectin chamber, 10 ng / ml in PBS, for 30 minutes at room temperature. Rinse once with PBS. Then place on plates COS cells at 2 - 3 x 105 cells per chamber in 1.5 ml of growth medium. Incubate overnight at 37 ° C. On day 1 for each sample, prepare 0.5 ml of a solution of 66 μg / ml DEAE-dextran, 66 μM chloroquine and 4 μg DNA in serum-free DME. For each set, a positive control is prepared, for example, of hulL-170-FLAG cDNA at 1 and 1/200 dilution, and a negative imitation. Rinse the cells with serum-free DME. Add the DNA solution and incubate for 5 hours at 37 ° C. Remove the medium and add 0.5 ml DMSO 10% in DME for 2.5 minutes. Stir and wash once with DME. Add 1.5 ml growth medium and incubate overnight. On day 2, change the medium. On days 3 or 4 the cells are fixed and stained. Rinse the cells twice with Saline with Hank's Buffer (HBSS) and fix in paraformaldehyde 4% (PFA) / glucose for 5 minutes. Wash 3 X with HBSS. The slides can be stored at -80 ° C after all the liquid is removed. For each camera, incubations of 0.5 ml are carried out in the following manner. Add HBSS / saponin (0.1%) with 32 μl / ml of 1 M NaN3 for 20 minutes. The cells are then washed with HBSS / 1 X saponin. Soluble antibody is added to the cells and incubated for 30 minutes. Wash the cells twice with HBSS / saponin. Add second antibody, for example, vector anti-mouse antibody, at 1/200 dilution, and incubate for 30 minutes. Prepare ELISA solution, for example, Vector Elite ABC radish horseradish peroxidase solution, and pre-incubate for 30 minutes. Use, for example, 1 drop of solution A (avidin) and 1 drop of solution B (biotin) per 2.5 ml HBSS / saponin. Wash the cells twice with HBSS / saponin. Add ABC HRP solution and incubate for 30 minutes. Wash the cells twice with HBSS, wash second for 2 minutes, bringing the cells closer. Then add diaminobenzoic acid Vector (DAB) for 5 to 10 minutes. Use 2 drops of buffer plus 4 DAB drops plus 2 drops of H2O2 per 5 ml of distilled glass water. Carefully remove the chamber and rinse the slide in water. Air dry for a few minutes, then add 1 drop of Crystal Mount and cover strip. Bake for 5 minutes at 85-90 ° C. Alternatively, the labeled ligand is used to purify by affinity or to classify cells expressing the receptor. See, for example, Sambrook et al., Or Ausubel et al. All references cited herein are incorporated herein by reference to the same extent as if each publication or individual patent application were specifically and individually indicated to be incorporated by reference. Many modifications and variations of this invention can be made, without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments described herein are offered by way of example only, and the invention should be limited only by the terms of the appended claims, together with the full scope of equivalents to which those claims are entitled.

PRESENTATION OF SEQUENCES SEQ ID NO: 1 is primate IL-172 nucleic acid sequence. SEQ ID NO: 2 is primate IL-172 polypeptide sequence. SEQ ID NO: 3 is murine IL-172 nucleic acid sequence. SEQ ID NO: 4 is murine IL-172 polypeptide sequence. SEQ ID NO: 5 is primate IL-173 nucleic acid sequence. SEQ ID NO: 6 is primate IL-173 polypeptide sequence. SEQ ID NO: 7 is IL-173 supplemental primate nucleic acid sequence. SEQ ID NO: 8 is IL-173 supplemental primate polypeptide sequence.

SEQ ID NO: 9 is murine IL-173 nucleic acid sequence. SEQ ID NO: 10 is murine IL-173 polypeptide sequence. SEQ ID NO: 11 is IL-173 supplemental murine nucleic acid sequence. SEQ ID NO: 12 is IL-173 supplemental murine polypeptide sequence.

SEQ ID NO: 13 is primate IL-174 nucleic acid sequence. SEQ ID NO: 14 is primate IL-174 polypeptide sequence. SEQ ID NO: 15 is murine IL-174 nucleic acid sequence. SEQ ID NO: 16 is murine IL-174 polypeptide sequence. SEQ ID NO: 17 is IL-174 nucleic acid sequence of murine supplementary. SEQ ID NO: 18 is IL-174 supplemental murine polypeptide sequence.

SEQ ID NO: 19 is primate IUPAC IL-171 nucleic acid sequence.

SEQ ID NO: 20 is primate IL-171 nucleic acid sequence. SEQ ID NO: 21 is primate IL-171 polypeptide sequence. SEQ ID NO: 22 is IL-171 supplemental primate nucleic acid sequence. SEQ ID NO: 23 is IL-171 supplemental primate polypeptide sequence.

SEQ ID NO: 24 is primate IUPAC IL-175 nucleic acid sequence.

SEQ ID NO: 25 is primate IL-175 nucleic acid sequence. SEQ ID NO: 26 is primate IL-175 polypeptide sequence. SEQ ID NO: 27 is primate IL-176 nucleic acid sequence. SEQ ID NO: 28 is primate IL-176 polypeptide sequence. SEQ ID NO: 29 is primate IL-177 nucleic acid sequence. SEQ ID NO: 30 is primate IL-177 polypeptide sequence. SEQ ID NO: 31 is a rat CTLA-8 polypeptide sequence. SEQ ID NO: 32 is mouse CTLA-8 polypeptide sequence. SEQ ID NO: 33 is primate CTLA-8 polypeptide sequence. SEQ ID NO: 34 is a viral CTLA-8 polypeptide sequence.

LIST OF SEQUENCES < 110 > Schering Corporation < 120 > Purified mammalian cytokines; Related reagents and methods < 130 > DX0917K < 140 > < 141 > < 150 > USSN 09 / 228,822 < 151 > 1999-01-11 < 160 > 34 < 170 > Patentln Ver. 2.1 < 210 > 1 < 211 > 543 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (1) .. (540) < 220 > < 221 > peptide mat_ < 222 > (61) .. (540) < 400 > 1 atg gac tgg ect cae aac ctg ctg ttt ett ett acc att tec ate ttc 48 Met Asp Trp Pro His Asn Leu Leu Phe Leu Leu Thr lie Ser lie Phe -20 -15 -10 -5 ctg ggg ctg ggc cag ecc agg age ecc aaa age aag agg aag ggg ca 96 Leu Gly Leu Gly Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln -1 1 5 10 ggg cgg ect ggg ecc ctg gtc ect ggc ect cae cag gtg cea ctg gac 144 Gly Arg Pro Gly Pro Leu Val Pro Gly Pro His Gln Val Pro Leu Asp 15 20 25 ctg gtg tea cgg atg aaa ceg tat gcc cgc atg gag gag tat gag agg 192 Leu Val Ser Arg Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg 30 35 40 aac ate gag gag atg gtg gcc cag ctg agg aac age tea gag ctg gcc 240 Asn lie Glu Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing 45 50 55 60 cag aga aag tgt gag gtc aac ttg cag ctg tgg atg tec aac aag agg 288 - | Q Gln Arg Lys Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg 65 70 75 age ctg tet ecc tgg ggc tac age ate aac falls gac ecc age ct 336 Ser Leu Ser Pro Trp Gly Tyr Ser lie Asn His Asp Pro Ser Arg lie 80 85 90 ecc gtg gac ctg ceg gag gca cgg tgc ctg tgt ctg ggc tgt gtg aac 384 Pro Val Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn 95 100 105 15 ecc ttc acc atg cag gag gac cgc age atg gtg age gtg ceg gtg ttc 432 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 110 115 120 age cag gtt ect gtg cgc cgc cgc etc tgc ceg cea ceg ecc cgc here 480 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr 125 130 135 140 ggg ect tgc cgc cag cgc gca gtc atg gag acc ate gct gtg ggc tgc 528 Gly Pro Cys Arg Gln Arg Ala Val Met Glu Thr lie Wing Val Gly Cys 145 150 155 acc tgc ate ttc tga 543 Thr Cys lie Phe 160 < 210 > 2 < 211 > 180 < 212 > PRT < 213 > primate < 400 > 2 Met Asp Trp Pro His Asn Leu Leu Phe Leu Leu Thr lie Ser lie Phe -20 -15 -10 -5 . { - Leu Gly Leu Gly Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln -1 1 5 10 Gly Arg Pro Gly Pro Leu Val Pro Gly Pro His Gln Val Pro Leu Asp 15 20 25 Leu Val Ser Arg Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg 30 35 40 Asn lie Glu Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing 45 50 55 60 Gln Arg Lys Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg '"65 70 75 Ser Leu Ser Pro Trp Gly Tyr Ser lie Asn His Asp Pro Ser Arg lie 80 85 90 Pro Val Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn 95 100 105 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 110 115 120 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr 5 125 130 135 140 Gly Pro Cys Arg Gln Arg Wing Val Met Glu Thr lie Wing Val Gly Cys 145 150 155 Thr Cys lie Phe 160 < 210 > 3 < 211 > 543 < 212 > DNA < 213 > rodent 0 < 220 > < 221 > CDS < 222 > (1) .. (540) < 220 > < 221 > peptide mat_ < 222 > (67) .. (540) < 400 > 3 atg gac tgg ceg falls age ctg etc ttc etc ctg gcc ate ate ttc tec 48 Met Asp Trp Pro His Ser Leu Leu Phe Leu Leu Ala lie Ser lie Phe -20 -15 -10 ctg gcg cea age falls ecc cgg aac acc aaa ggc aaa aga aaa ggg caa 96 Leu Ala Pro Ser His Pro Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln -5 -1 1 5 10 ggg agg ecc agt ecc ttg gcc ect ggg ect cat cag gtg ceg ctg gac 144 Gly Arg Pro Pro Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp 15 20 25 ctg gtg tet cga gta aag ecc tac gct cga atg gaa gag tat gag cgg 192 Leu Val Ser Arg Val Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg 30 35 40 aac ett ggg gag atg gtg gcc cag ctg agg aac age tec gag cea gcc 240 Asn Leu Gly Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Pro Wing 45 50 55 aag aaag aaa tgt gaa gtc aat cta cag ctg tgg ttg tec aac aag agg 288 Lys Lys Cys Glu Val Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg 60 65 70 age ctg tec ce tgg ggc tac age ate aac falls gac ecc age cgc ate 336 Ser Leu Ser Pro Trp Gly Tyr Ser lie Asn His Asp Pro Ser Arg lie 75 80 85 90 ect gcg gac ttg ecc gag gcg cgg tgc cta tgt ttg ggt tgc gtg aat 384 Pro Wing Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn 95 100 105 ecc ttc acc atg cag gag gac cgt age atg gtg age gtg cea gtg ttc 432 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 110 115 120 age cag gtg ceg gtg cgc cgc cgc etc tgt ect cact ect ect cctcct 480 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro 125 130 135 ggg ecc tgc cgc cag cgt gtc gtc atg gag acc ate gct gtg ggt tgc 528 Gly Pro Cys Arg Gln Arg Val Val Met Glu Thr lie Wing Val Gly Cys 140 145 150 acc tgc ate ttc tga 543 Thr Cys lie Phe 155 < 210 > 4 < 211 > 180 < 212 > PRT < 213 > rodent < 400 > 4 Met Asp Trp Pro His Ser Leu Leu Phe Leu Leu Ala lie Ser lie Phe -20 -15 -10 Leu Ala Pro Ser His Pro Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln -5 -1 1 5 10 Gly Arg Pro Pro Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp 15 20 25 Leu Val Ser Arg Val Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg 30 35 40 Asn Leu Gly Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Pro Wing 45 50 55 Lys Lys Cys Glu Val Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg 60 65 70 Ser Leu Ser Pro Trp Gly Tyr Ser lie Asn His Asp Pro Ser Arg lie 75 80 85 90 Pro Wing Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn 95 100 105 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 110 115 120 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro 125 130 135 Gly Pro Cys Arg Gln Arg Val Val Met Glu Thr lie Wing Val Gly Cys 140 145 150 Thr Cys lie Phe 155 < 210 > 5 < 211 > 454 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (1) .. (453) < 400 > 5 tgc gcg gac cgg ceg gag gag cta cg gag cag ctg tac ggg cgc ctg 48 Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu 1 5 10 15 gcg gcc ggc gtg etc agt gcc ttc falls falls acg ctg cag ctg ggg ceg 96 Wing Wing Gly Val Leu Ser Wing Phe His His Thr Leu Gln Leu Gly Pro 20 25 30 cgt gag cag gcg cgc aac gcg age tgc ceg gca ggc ggc agg ecc gcc 144 Arg Glu Gln Wing Arg Asn Wing Being Cys Pro Wing Gly Gly Arg Pro Wing 35 40 45 gac cgc cgc ttc cgg acg ecc acc aac ctc cgc age gtg teg ecc tgg 192 Asp Arg Arg Phe Arg Thr Pro Thr Asn Leu Arg Ser Val Ser Pro Trp 50 55 60 gcc tac aga ate tec tac gac cec gcg agg tac ecc agg tac ctg ect 240 Wing Tyr Arg lie Ser Tyr Asp Pro Wing Arg Tyr Pro Arg Tyr Leu Pro 65 70 75 80 gaa gcc tac tgc ctg tgc cgg ggc tgc ctg acc ggg ctg ttc ggc gag 288 Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr Gly Leu Phe Gly Glu 85 90 95 gag gac gtg cgc ttc cgc age gcc ect gtc tac atg ecc acc gtc gtc 336 Glu Asp Val Arg Phe Arg Ser Wing Pro Val Tyr Met Pro Thr Val Val 100 105 110 ctc cgc ccg ccc acccc ccc gcc cc gcc gcc gcc gcc ccc acc gag 384 Leu Arg Arg Thr Pro Ala C ys Wing Gly Gly Arg Ser Val Tyr Thr Glu 115 120 125 gcc tac gtc acc ate ecc gtg ggc tgc acc tgc gtc ecc gag ceg gag 432 Wing Tyr Val Thr lie Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu 130 135 140 aag gac gca gac age ate aac t 454 Lys Asp Ala Asp Ser lie Asn 145 150 < 210 > 6 < 211 > 151 < 212 > PRT < 213 > primate < 400 > 6 Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu 1 5 10 15 Ala Ala Gly Val Leu Ser Ala Phe His His Thr Leu Gln Leu Gly Pro 20 25 30 Arg Glu Gln Ala Arg Asn Ala Ser Cys Pro Wing Gly Gly Arg Pro Wing 35 40 45 Asp Arg Arg Phe Arg Thr Pro Thr Asn Leu Arg Ser Val Ser Pro Trp 50 55 60 Wing Tyr Arg lie Ser Tyr Asp Pro Wing Arg Tyr Pro Arg Tyr Leu Pro 65 70 75 80 Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr Gly Leu Phe Gly Glu 85 90 95 Glu Asp Val Arg Phe Arg Ser Wing Pro Val Tyr Met Pro Thr Val Val 100 105 110 Leu Arg Arg Thr Pro Wing Cys Wing Gly Gly Arg Ser Val Tyr Thr Glu 115 120 125 Wing Tyr Val Thr lie Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu 130 135 140 Lys Asp Wing Asp Ser lie Asn 145 150 < 210 > 7 < 211 > 1385 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (59) .. (664) < 220 > < 221 > peptide mat_ < 222 > (110) .. (664) < 400 > 7 gcccgggcag gtggcgacct cgctcagtcg gcttctcggt ccg gcctetgg gcggcgcg cgct ggcgcg gcgcgc gcg ccg ctg ccg gcg gcg gcg gcg ctg gcg ctg cg ggc gcg 106 Met Leu Val Wing Gly Phe Leu Leu Wing Le Pro Pro Ser Wing Trp Wing -15 -10 -5 ggc gcc cg agg gcg ggc agg cgc ecc gcg cgg ceg cgg ggc tgc gcg 154 Gly Ala Pro Arg Ala Gly Arg Arg Pro Ala Arg Pro Arg Gly Cys Ala -1 1 5 10 15 gac cgg ceg gag gag cta ctg gag cag ctg tac ggg cgc ctg gcg gcc 202 Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu Ala Wing 20 25 30 ggc gtg etc agt gcc ttc falls falls acg ctg cag ctg ggg ceg cgt gag 250 Gly Val Leu Ser Wing Phe His His Thr Leu Gln Leu Gly Pro Arg Glu 35 40 45 cag gcg cgc aac gcg age tgc ceg gca ggg ggc agg ecc gcc gac cgc 298 Gln Wing Arg Asn Wing Ser Cys Pro Wing Gly Gly Arg Pro Wing Asp Arg 50 55 60 cgc ttc cgg ceg ecc acc aac ctc cgc age gtg teg ecc tgg gcc tac 346 Arg Phe Arg Pro Pro Thr Asn Leu Arg Ser Val Ser Pro Trp Wing Tyr 65 70 75 aga ate tec tac gac ceg gcg agg tac ecc agg tac ctg ect gaa gcc 394 Arg l ie Ser Tyr Asp Pro Wing Arg Tyr Pro Arg Tyr Leu Pro Glu Wing 80 85 90 95 tac tgc ctg tgc cgg ggc tgc ctg acc ggg ctg ttc ggc gag gag gac 442 Tyr Cys Leu Cys Arg Gly Cys Leu Thr Gly Leu Phe Gly Glu Glu Asp 100 105 110 gtg cgc ttc cgc age gcc ect gtc tac atg ecc acc gtc gtc cgc cgc 490 Val Arg Phe Arg Ser Wing Pro Val Tyr Met Pro Thr Val Val Leu Arg 115 120 125 cgc acc ecc gcc tgc gcc ggc ggc cgt tec gtc tac acc gag gcc tac 538 Arg Thr Pro Wing Cys Wing Gly Gly Arg Ser Val Tyr Thr Glu Wing Tyr 130 135 140 gtc acc ate ecc gtg ggc tgc acc tgc gtc ecc gag ceg gag aag gac 586 Val Thr lie Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu Lys Asp 145 150 155 gca gac age ate aac tec age ate gac aaa cag ggc gcc aag etc ctg 634 Wing Asp Ser lie Asn Ser Ser lie Asp Lys Gln Gly Ala Lys Leu Leu 160 165 170 175 ctg ggc ecc aac gac gcg ecc gct gcc ecc tgaggccggt cctgccccgg 684 Leu Gly Pro Asn Asp Ala Pro Wing Gly Pro 180 185 gaggtctccc cggcccgcat cccgaggcgc ccaagctgga gccgcctgga gggctcggtc 744 ggcgacctct gaagag agtg caccgagcaa accaagtgcc ggagcaccag cgccgccttt 804 ccatggagac tcgtaagcag cttcatctga cacgggcatc cctggcttgc ttttagctac 864 aagcaagcag cgtggctgga agctgatggg aaacgacccg gcacgggcat cctgtgtgcg 924 gcccgcatgg agggtttgga aaagttcacg gaggctccct gaggagcctc tcagatcggc 984 tgetgegggt gcagggcgtg actcaccgct gggtgcttgc gggaegeata caaagagata 1044 agcaatctaa tgctttttaa aaataataat aagtatagcg actatatacc tacttttaaa 1104 atcaactgtt ttgaatagag gcagagctat tttatattat caaatgagag ctactctgtt 1164 acatttetta acatataaac atcgtttttt acttcttctg gtagaatttt ttaaagcata 1224 attggaatcc ttggataaat tttgtagctg gtacactctg gcctgggtct ctgaattcag 1284 cctgtcaccg atggctgact gatgaaatgg acacgtctca tctgacccac tcttccttcc 1344 actgaaggtc ttcacgggcc tccaggcctc gtgccgaatt c 1385 < 210 > 8 < 211 > 202 < 212 > PRT < 213 > primate < 400 > 8 Met Leu Val Wing Gly Phe Leu Leu Wing Leu Pro Pro Wing Trp Wing Wing -15 -10 -5 Gly Wing Pro Wing Arg Wing Arg Arg Pro Wing Wing Arg Pro Arg Gly Wing Cys -1 1 5 10 15 Asp Arg Pro Glu Glu Leu Leu Glu Gln Leu Tyr Gly Arg Leu Wing Ala 20 25 30 Gly Val Leu Be Ala Phe His His Thr Leu Gln Leu Gly Pro Arg Glu 35 40 45 Gln Wing Arg Asn Wing Cys Pro Wing Gly Gly Arg Pro Wing Asp Arg 50 55 60 Arg Phe Arg Pro Pro Thr Asn Leu Arg Ser Val Ser Pro Trp Ala Tyr 65 70 75 Arg lie Ser Tyr Asp Pro Ala Arg Tyr Pro Arg Tyr Leu Pro Glu Wing 80 85 90 95 Tyr Cys Leu Cys Arg Gly Cys Leu Thr Gly Leu Phe Gly Glu Glu Asp 100 105 110 Val Arg Phe Arg Ser Wing Pro Val Tyr Met Pro Thr Val Val Leu Arg 115 120 125 Arg Thr Pro Wing Cys Wing Gly Gly Arg Ser Val Tyr Thr Glu Wing Tyr 130 135 140 Val Thr lie Pro Val Gly Cys Thr Cys Val Pro Glu Pro Glu Lys Asp 145 150 155 Wing Asp Ser lie Asn Ser Ser lie Asp Lys Gln Gly Wing Lys Leu Leu 160 165 170 175 Leu Gly Pro Asn Asp Ala Pro Wing Gly Pro 180 185 < 210 > 9 < 211 > 133 < 212 > DNA < 213 > rodent < 220 > < 221 > CDS < 222 > (1) .. (132) < 400 > 9 ttt ceg aga tac ctg ecc gaa gcc tac tgc ctg tgc cga ggc tgt ctg 48 Phe Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu '1 5 10 15 acc ggg etc tac ggt gag gac gac tcc cgc ttt cgc age gca ecc gtc 96 Thr Gly Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Wing Pro Val 20 25 30 ttc tet ceg gcg gtg gtg ctg cgg cgc acg gcg gcc t 133 Phe Ser Pro Ala Val Val Leu Arg Arg Thr Ala Ala 35 40 < 210 > 10 < 211 > 44 < 212 > PRT < 213 > rodent < 400 > 10 Phe Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu 1 5 10 15 Thr Gly Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Wing Pro Val 20 25 30 Phe Ser Pro Wing Val Val Leu Arg Arg Thr Ala Ala 35 40 < 210 > 11 < 211 > 1143 < 212 > DNA < 213 > rodent < 220 > < 221 > CDS < 222 > (1) .. (615) < 220 > < 221 > peptide mat_ < 222 > (73) .. (615) < 400 > 11 atg ttg ggg here ctg gtc tgg atg etc etc gtc ggc ttc ctg ctg gca 48 Met Leu Gly Thr Leu Val Trp Met Leu Leu Val Gly Phe Leu Leu Wing -20 -15 -10 ctg gcg ceg ggc cgc gcg gcg ggc gcg ctg agg acc ggg agg cgc ceg 96 Leu Ala Pro Gly Arg Ala Wing Gly Ala Leu Arg Thr Gly Arg Arg Pro -5 -1 1 5 gcg cgg ceg cgg gac tgc gcg gac cgg cea gag gag etc ctg gag cag 144 Wing Arg Pro Arg Asp Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln 10 15 20 ctg tac ggg cgg cg gcg gcg ggc gtg etc age gcc ttc falls falls acg 192 Leu Tyr Gly Arg Leu Wing Wing Gly Val Leu Ser Wing Phe His His Thr 25 30 35 40 ctg cag etc ggg ceg cgc gag cag gcg cgc aat gcc age tgc ceg gcc 240 Leu Gln Leu Gly Pro Arg Glu Gln Wing Arg Asn Wing Cys Pro Wing 45 50 55 999 ggc agg gcc gcc gac cgc cgc ttc cgg cea ecc acc aac ctg cgc 288 Gly Gly Arg Wing Wing Asp Arg Arg Phe Arg Pro Pro Thr Asn Leu Arg 60 65 70 age gtg teg ecc tgg gcg tac agg att tec tac gac ect gct cgc ttt 336 Ser Val Ser Pro Trp Wing Tyr Arg lie Ser Tyr Asp Pro Wing Arg Phe 75 80 85 ceg agg tac ctg ecc gaa gcc tac tgc ctg tgc cga ggc tgc ctg acc 384 Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr 90 95 100 ggg etc tac ggg gag gac gac ttc cgc ttt cgc age here ecc gtc ttc 432 Gly Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Thr Pro Val Phe 105 110 115 120 tet cea gcc gtg gtg ctg cgg cgc here gcg gcc tgc gcg ggc ggc cgc 480 Ser Pro Ala Val Val Leu Arg Arg Thr Ala Ala Cys Ala Gly Gly Arg 125 130 135 tet gtg tac gcc gaa fall tac ate acc ate ceg gtg ggc tgc acc tgc 528 Ser Val Tyr Ala Glu His Tyr He Thr He Pro Val Gly Cys Thr Cys 140 145 150 gtg ecc gag ceg gac aag tec gcg gac agt gcg aac tec age atg gac 576 Val Pro Glu Pro Asp Lys Ser Wing Asp Ser Wing Asn Ser Ser Met Asp 155 160 165 aag ctg ctg ctg ggg ecc gcc gac agg ect gcg ggg cgc tgatgccggg 625 Lys Leu Leu Glu Pro Pro Wing Asp Arg Pro Ala Gly Arg 170 175 180 gactgcccgc catggcccag cttcctgcat gcatcaggtc ccctggccct gacaaaaccc 685 accccatgat ccctggccgc tgcctaattt ttccaaaagg acagetacat aagctttaaa 745 aaagtagaca tatatttttc etacatatet acaactattt tgaatagtgg cagaaactat 805 gtaatttaga ttteatatta gcaagcatgt tgtttttaaa cttctttgat atacaageac 865 atcacacaca tcccgttttc ctctagtagg attettgagt gcataattgt agtgetcaga 925 tgaa cttcct tctgctgcac tgtgccctgt ccctgagtct ctcctgtggc ccaagcttac 985 taaggtgata atgagtgctc cggatetggg cacctaaggt ctccaggtcc ctggagaggg 1045 agggatgtgg gggggctagg aaccaagcgc ccctttgttc tttagcttat ggatggtctt 1105 aactttataa agattaaagt ttttggtgtt attctttc 1143 < 210 > 12 < 211 > 205 < 212 > PRT < 213 > rodent < 400 > 12 5 Met Leu Gly Thr Leu Val Trp Met Leu Leu Val Gly Phe Leu Leu Wing -20 -15 -10 Leu Wing Pro Gly Arg Wing Wing Gly Wing Leu Arg Thr Gly Arg Arg Pro -5 -1 1 5 Wing Arg Pro Arg Asp Cys Wing Asp Arg Pro Glu Glu Leu Leu Glu Gln 10 15 20 Leu Tyr Gly Arg Leu Wing Wing Gly Val Leu Ser Wing Phe His His Thr 25 30 35 40 - | Q Leu Gln Leu Gly Pro Arg Glu Gln Wing Arg Asn Wing Being Cys Pro Wing 45 50 55 Gly Gly Arg Wing Wing Asp Arg Arg Phe Arg Pro Pro Thr Asn Leu Arg 60 65 70 Ser Val Ser Pro Trp Wing Tyr Arg He Be Tyr Asp Pro Wing Arg Phe 75 80 85 Pro Arg Tyr Leu Pro Glu Wing Tyr Cys Leu Cys Arg Gly Cys Leu Thr 90 95 100 AC Gly Leu Tyr Gly Glu Glu Asp Phe Arg Phe Arg Ser Thr Pro Val Phe 105 110 115 120 Ser Pro Ala Val Val Leu Arg Arg Thr Ala Ala Cys Ala Gly Gly Arg 125 130 135 Ser Val Tyr Ala Glu His Tyr He Thr He Pro Val Gly Cys Thr Cys 140 145 150 Val Pro Glu Pro Asp Lys Ser Wing Asp Ser Ala Asn Ser Ser Met Asp 155 160 165 Lys Leu Leu Gly Pro Wing Asp Arg Pro Wing Gly Arg 0 170 175 180 < 210 > 13 < 211 > 504 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (19) .. (501) < 220 > < 221 > peptide mat_ < 222 > (67) .. (501) < 400 > 13 tgagtgtgca gtgccagc atg tac cag gtg gtt gca ttc ttg gca atg gtc 51 Met Tyr Gln Val Val Ala Phe Leu Ala Met Val -15 -10 atg gga acc falls age tagg tgg tcc ecc age tgc ecc age aaa 99 Met Gly Thr His Thr Tyr Ser His Trp Pro Ser Cys Cys Pro Ser Lys -5 -1 1 5 10 ggg cag gac acc tet gag gag ctg ctg agg tgg age act gtg ect gtg 147 Gly Gln Asp Thr Ser Glu Glu Leu Leu Arg Trp Ser Thr Val Pro Val 15 20 25 ect ecc cta gag ect gct agg ecc aac cgc cae ce gag tec tgt agg 195 Pro Pro Leu Glu Pro Wing Arg Pro Asn Arg His Pro Glu Ser Cys Arg 30 35 40 gcc agt gaa gat gga ecc etc aac age agg gcc ate tec ecc tgg aga 243 Wing Ser Glu Asp Gly Pro Leu Asn Ser Arg Ala He Ser Pro Trp Arg 45 50 55 tat gag ttg gac aga gac ttg aac cgg etc ecc cag gac ctg tac falls 291 Tyr Glu Leu Asp Arg Asp Leu Asn Arg Leu Pro Gln Asp Leu Tyr His 60 65 70 75 gcc cgt tgc ctg tgc ceg falls tgc gtc age cta cag here ggc tec falls 339 Wing Arg Cys Leu Cys Pro His Cys Val Ser Leu Gln Thr Gly Ser His 80 85 90 atg gac ecc cgg ggc aac teg gag ctg etc tac falls aac cag act gtc 387 Met Asp Pro Arg Gly Asn Ser Glu Leu Leu Tyr His Asn Gln Thr Val 95 100 105 ttc tac cgg cgg cea tgc cat ggc gag aag ggc acc falls aag ggc tac 435 Phe Tyr Arg Arg Pro Cys His Gly Glu Lys Gly Thr His Lys Gly Tyr 110 115 120 tgc ctg gag cgc agg ctg tac cgt gtt tec tta gct tgt gtg tgt gtg 483 Cys Leu Glu Arg Arg Leu Tyr Arg Val Ser Leu Ala Cys Val Cys Val 125 130 135 cgg ecc cgt gtg atg ggc tag 504 Arg Pro Arg Val Met Gly 140 145 < 210 > 14 < 211 > 161 < 212 > PRT < 213 > primate < 400 > 14 Met Tyr Gln Val Val Ala Phe Leu Ala Met Val Met Gly Thr His Thr -15 -10 -5 -1 Tyr Ser His Trp Pro Ser Cys Cys Pro Ser Lys Gly Gln Asp Thr Ser 1 5 10 15 Glu Glu Leu Leu Arg Trp Ser Thr Val Pro Pro Pro Leu Glu Pro 20 25 30 Wing Arg Pro Asn Arg His Pro Glu Ser Cys Arg Wing Ser Glu Asp Gly 35 40 45 Pro Leu Asn Ser Arg Wing He Ser Pro Trp Arg Tyr Glu Leu Asp Arg 50 55 60 Asp Leu Asn Arg Leu Pro Gln Asp Leu Tyr His Wing Arg Cys Leu Cys 65 70 75 80 Pro His Cys Val Ser Leu Gln Thr Gly Ser His Met Asp Pro Arg Gly 85 90 95 Asn Ser Glu Leu Leu Tyr His Asn Gln Thr Val Phe Tyr Arg Arg Pro 100 105 110 Cys His Gly Glu Lys Gly Thr His Lys Gly Tyr Cys Leu Glu Arg Arg 115 120 125 Leu Tyr Arg Val Ser Leu Ala Cys Val Cys Val Arg Pro Arg Val Met 130 135 140 Gly 145 < 210 > 15 < 211 > 620 < 212 > DNA < 213 > rodent < 220 > < 221 > CDS < 222 > (1) - • (432) < 400 > 15 cgg drops agg cgg drops aaa gcc cgg aga gtg gct gaa gtg gag etc tgc 48 Arg His Arg Arg His Lys Ala Arg Arg Val Ala Glu Val Glu Leu Cys 1 5 10 15 ate tgt ate ecc ecc aga gcc tet gag cea cae cea cea cgc aga ate 96 He Cys He Pro Pro Arg Ala Ser Glu Pro His Pro Pro Arg Arg He 20 25 30 ctg cag ggc cag ca gga tgg ect etc aac age agg gcc ate tet ect 144 Leu Gln Gly Gln Gln Gly Trp Pro Leu Asn Ser Arg Ala He Ser Pro 35 40 45 tgg age tat gag ttg gac agg gac ttg aat cgg gtc ecc cag gac tgg 192 c Trp Ser Tyr Glu Leu Asp Arg Asp Leu Asn Arg Val Pro Gln Asp Trp 50 55 60 tac drops gct cga tgc ctg tgc cea drops tgc gtc acg cta cag here ggc 240 Tyr His Wing Arg Cys Leu Cys Pro His Cys Val Thr Leu Gln Thr Gly 65 70 75 80 tec falls atg gac ceg ctg ggc aac tec gtc cea ett tac falls aac cag 288 Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn Gln 85 90 95"10 acg gtc ttc tac cgg cgg cgc atc gcg agg aag gta ecc ate gcc 336 Thr Val Phe Tyr Arg Arg Pro Cys Met Wing Arg Lys Val Pro lie Wing 100 105 110 gct act gct tgg age gg ggt cta ceg agt etc gtc gtc gtg tgt tgt 384 Wing Thr Wing Trp Wing Wing Gly Leu Pro Leu Leu Gly Leu Cys Val 115 120 125 tgt gcg gcg ggt ggt cat ggc tta gtc tg atg etc acc ate tgc ctg agg 432 <; | 5 Cys Ala Ala Gly Leu Pro Gly His Val Thr Met Leu Leu Arg Cys He 130135140 tgaatgccgg gtgggagaga gggccaggtg tacatcacct gccaatgcgg gccgggttca 492 gcctacctga agcctgcaaa agcagcaggt cccgggacag gatggagact tggggagaaa 552 tctgactttt gcactttttg gagcattttg ggaagagcag gttcgcttgt gctgtagaga 612 tgctgttg 620 < 210 > 16 0 < 211 > 144 < 212 > PRT < 213 > rodent < 400 > 16 Arg His Arg Arg His Lys Wing Arg Arg Val Wing Glu Val Glu Leu Cys 1 5 10 15 He Cys He Pro Pro Arg Wing Ser Glu Pro His Pro Pro Arg Arg He 20 25 30 Leu Gln Gly Gln Gln Gly Trp Pro Leu Asn Be Arg Ala Be Ser Pro 35 40 45 Trp Ser Tyr Glu Leu Asp Arg Asp Leu Asn Arg Val Pro Gln Asp Trp 50 55 60 Tyr His Wing Arg Cys Leu Cys Pro His Cys Val Thr Leu Gln Thr Gly 65 70 75 80 Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn Gln 85 90 95 Thr Val Phe Tyr Arg Arg Pro Cys Met Wing Arg Lys Val Pro He Wing 100 105 110 Wing Thr Wing Trp Wing Wing Gly Leu Pro Ser Leu Leu Gly Leu Cys Val 115 120 125 Cys Wing Wing Pro Gly His Gly Leu Val Met Leu Thr He Cys Leu Arg 130 135 140 < 210 > 17 < 211 > 985 < 212 > DNA < 213 > rodent < 220 > < 221 > CDS < 222 > (1) .. (507) < 220 > < 221 > peptide mat_ < 222 > (49) .. (507) < 400 > 17 atg tac cag gct gtt gt ttc ttg gca atg ate gtg gga acc fall 48 Met Tyr Gln Ala Val Ala Phe Leu Ala Met He Val Gly Thr His Thr -15 -10 -5 -1 gtc age ttg cgg ate cag gag ggc tgc agt falls ttg ecc age tgc tgc 96 Val Ser Leu Arg He Gln Glu Gly Cys Ser His Leu Pro Ser Cys Cys 1 5 10 15 ecc age aaa gag caa gaa ecc ceg gag gag tgg ctg aag tgg age tet 144 Pro Ser Lys Glu Gln Glu Pro Pro Glu Glu Trp Leu Lys Trp Ser Ser 20 25 30 gca tet gtg tec ecc cea gag ect ctg age falls acc fall falls gca gaa 192 Wing Ser Val Ser Pro Pro Glu Pro Leu Ser His Thr His His Wing Glu 35 40 45 tec tgc agg gcc age aag gat ggc ecc etc aac age agg gcc ate tet 240 Ser Cys Arg Ala Ser Lys Asp Gly Pro Leu Asn Ser Arg Ala He Ser 50 55 60 ect tgg age tat gag ttg gac agg gac ttg aat cgg gtc ecc cag gac 288 Pro Trp Ser Tyr Glu Leu Asp Arg Asp Leu Asn Arg Val Pro Gln Asp 65 70 75 80 ctg tac drops gct cga tgc ctg tgc cea falls tgc gtc age cta cag here 336 Leu Tyr His Ala Arg Cys Leu Cys Pro His Cys Val Ser Leu Gln Thr 85 90 95 ggc tec falls atg gac ceg ctg ggc aac tec gtc cea ett tac falls aac 384 Gly Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn 100 105 110 cag acg gtc ttc tac cgg cgg cg cea tgc cat ggt gag gaa ggt acc cat 432 Gln Thr Val Phe Tyr Arg Arg Pro Cys His Gly Glu Glu Gly Thr His 115 120 125 cgc cgc tac tgc ttg gag cgc agg etc tac cga gtc tec ttg gct tgt 480 Arg Arg Tyr Cys Leu Glu Arg Arg Leu Tyr Arg Val Ser Leu Ala Cys 130 135 140 gtg tgt gtg cgg ecc cgg gtc atg gct tagtcatget caccacctgc 527 Val Cys Val Arg Pro Arg Val Met Ala 145 150 ctgaggctga tgcccggttg ggagagaggg ccaggtgtac aatcaccttg ccaatgcggg 587 gccctccaaa ccgggttcaa gccctacctg aagcagcagg ctcccgggac aagatggagg 647 acttggggag aaactctgac ttttgcactt tttggaagca cttttgggaa ggageaggtt 707 ccgcttgtgc tgctagagga tgctgttgtg gcatttctac tcaggaacgg actccaaagg 767 cctgctgacc ctggaagcca tactcctggc tcctttcccc tgaatccccc aactcctggc 827 acaggcactt tctccacctc tccccctttg ccttttgttg tgtttgtttg tgcatgccaa 887 ctctgcgtgc agccaggtgt aattgccttg aaggat ggtt ctgaggtgaa agctgttatc 947 gaaagtgaag agatttatec aaataaacat ctgtgttt 985 < 210 > 18 < 211 > 169 < 212 > PRT < 213 > rodent < 400 > 18 Met Tyr Gln Ala Val Ala Phe Leu Ala Met He Val Gly Thr His Thr - 15 - 10 -5 - 1 Val Ser Leu Arg He Gln Glu Gly Cys Ser His Leu Pro Ser Cys Cys 1 5 10 15 Pro Ser Lys Glu Gln Glu Pro Pro Glu Glu Trp Leu Lys Trp Ser Ser 20 25 30 Wing Ser Val Ser Pro Pro Glu Pro Leu Ser His Thr His His Wing Glu 35 40 45 Ser Cys Arg Wing Ser Lys Asp Gly Pro Leu Asn Ser Arg Wing He Ser 50 55 60 Pro Trp Ser Tyr Glu Leu Asp Arg Asp Leu Asn Arg Val Pro Gln Asp 65 70 75 80 Leu Tyr His Wing Arg Cys Leu Cys Pro His Cys Val Ser Leu Gln Thr 85 90 95 Gly Ser His Met Asp Pro Leu Gly Asn Ser Val Pro Leu Tyr His Asn 100 105 110 Gln Thr Val Phe Tyr Arg Arg Pro Cys His Gly Glu Glu Gly Thr His 115 120 125 Arg Arg Tyr Cys Leu Glu Arg Arg Leu Tyr Arg Val Ser Leu Ala Cys 130 135 140 Val Cys Val Arg Pro Arg Val Met Wing 145 150 < 210 > 19 < 211 > 521 < 212 > DNA < 213 > primate < 220 > < 221 > feature mis_ < 222 > (1) .. (521) < 223 > note = "n can be a, c, g, or t" < 400 > 19 gacacggatg aggaccgcta tccacagaag ctggccttcg ccgagtgcct gtgcagaggc 60 tgtatcgatg cacggacggg ccgcgagaca gctgcgctca actccgtgcg gctgctccag 120 agcctgctgg tgctgcgccg ccggccctgc tcccgcgacg gctcggggct ccccacacct 180 ggggcctttg ccttccacac cgagttcatc cacgtccccg tcggctgcac ctgcgtgctg 240 ccccgttcaa gtgtgaccgc caaggccgtg gggcccttag ntgacaccgt gtgctcccca 300 gagggacccc tatttatggg aattatggta ttatatgctt cccacatact tggggctggc 360 atcccgngct gagacagccc cctgttctat tcagctatat ggggagaaga gtagactttc 420 agctaagtga aaagtgnaac gtgctgactg tctgctgtcg tnctactnat gctagcccga 480 gtgttcactc tgagcctgtt aaatataggc ggttatgtac c 521 <; 210 > 20 < 211 > 521 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (1) .. (369) < 220 > < 221 > feature mis_ < 222 > (281) < 223 > note = "nucleotides 281, 367, 437, 462, and 468 are indicating c; each may be alternatively a, g, ot, the translation amino acid depends on the genetic code <400> 20 gac acg gat gag gac cgc tat cea cag aag ctg gcc tcc gcc gag tgc 48 Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Wing Phe Wing Glu Cys 1 5 10 15 ctg tgc aga ggc tgt ate gat gca cgg acg ggc cgc gag here gct gcg 96 Leu Cys Arg Gly Cys He Asp Wing Arg Thr Gly Arg Glu Thr Wing Wing 20 25 30 etc aac tec gtg cgg ctg etc cag age ctg ctg gtg ctc cgc cgc cgg 144 Leu Asn Ser Val Arg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg Arg 35 40 45 ecc tgc tec cgc gac ggc teg ggg etc ecc here ect ggg gcc ttt gcc 192 Pro Cys Ser Arg Asp Gly Ser Gly Leu Pro Thr Pro Gly Ala Phe Wing 50 55 60 ttc falls acc gag ttc ate drops gtc ecc gtc ggc tgc acc tgc gtg ctg 240 phe His Thr Glu Phe He His Val Pro Val Gly Cys Thr Cys Val Leu 65 70 75 80 ecc cgt tea agt gtg acc gcc aag gcc ggg ggg ecc tta gct gac acc 288 Pro Arg Ser Val Thr Ala Lys Ala Val Gly Pro Leu Ala Asp Thr 85 90 95 gtg tgc tec cea gag gga ecc cta ttt atg gga att atg gta tta tat 336 Val Cys Ser Pro Glu Gly Pro Leu Phe Met Gly He Met Val Leu Tyr 100 105 110 gct tec falls ata et ggg gct ggc ate ceg cgc tgagacagee ccctgttcta 389 Ala Ser His He Leu Gly Ala Gly He Pro Arg 115 120 tteagetata tggggagaag agtagacttt cagetaagtg aaaagtgeaa cgtgctgact 449 gtctgctgtc gtcctactca tgctagcccg agtgttcact ctgagcctgt taaatatagg 509 cggttatgta ce 521 < 210 > 21 < 211 > 123 < 212 > PRT < 213 > primate < 400 > 21 Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Wing Phe Wing Glu Cys 1 5 10 15 Leu Cys Arg Gly Cys He Asp Wing Arg Thr Gly Arg Glu Thr Wing Wing 20 25 30 Leu Asn Ser Val Arg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg Arg 35 40 45 Pro Cys Ser Arg Asp Gly Ser Gly Leu Pro Thr Pro Gly Wing Phe Wing 50 55 60 Phe His Thr Glu Phe He His Val Val Val Gly Cys Thr Cys Val Leu 65 70 75 80 Pro Arg Ser Ser Val Thr Wing Lys Wing Val Gly Pro Leu Wing Asp Thr 85 90 95 Val Cys Ser Pro Glu Gly Pro Leu Phe Met Gly He Met Val Leu Tyr 100 105 110 Wing Ser His He Leu Gly Wing Gly He Pro Arg 115 120 < 210 > 22 < 211 > 1107 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (115) .. (705) < 220 > < 221 > peptide mat_ < 222 > (166) .. (705) < 400 > 22 gtgtggcctc aggtataaga gcggctgctg ccaggtgcat ggecaggtge aectgtggga 60 ttgccgccag gtgtgcaggc cgctccaagc ccagcctgcc ccgctgccgc cace atg 117 Met acg etc etc ecc ggc etc ctg ttt ctg acc tgg ctg falls here tgc ctg 165 Thr Leu Leu Pro Gly Leu Leu Phe Leu Thr Trp Leu His Thr Cys Leu -15 -10 -5 -1 gcc falls cat gac ecc tec etc agg ggg drops ecc falls agt falls ggt acc 213 Wing His His Asp Pro Ser Leu Arg Gly His Pro His Ser His Gly Thr 1 5 10 15 cea falls tgc tac teg gct gag gaa ctg ecc etc ggc cag gcc ecc cea 261 Pro His Cys Tyr Ser Ala Glu Glu Leu Pro Leu Gly Gln Ala Pro Pro 20 25 30 falls ctg ctg gct cga ggt gcc aag tgg ggg cag gct ttg ect gta gcc 309 His Leu Leu Ala Arg Gly Ala Lys Trp Gly Gln Ala Leu Pro Val Ala 35 40 45 ctg gtg tec age ctg gag gca gca age falls agg ggg agg falls gag agg 357 Leu Val Ser Ser Leu Glu Ala Ala Ser His Arg Gly Arg His Glu Arg 50 55 60 ecc tea gct acg acc cag tgc ceg gtg ctg cgg ceg gag gag gtg ttg 405 Pro Be Wing Thr Thr Gln Cys Pro Val Leu Arg Pro Glu Glu Val Leu 65 70 7 5 80 gag gca gac acc fall cag cgc tec ate tea ecc tgg aga tac cgt gtg 453 Glu Wing Asp Thr His Gln Arg Ser He Ser Pro Trp Arg Tyr Arg Val 85 90 95 gac acg gat gag gac cgc tat cea cag aag ctg gcc ttc gcc gag tgc 501 Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Ala Phe Ala Glu Cys 100 105 110 ctg tgc aga ggc tgt ate gat gca cgg acg ggc cgc gag here gct gcg 549 Leu Cys Arg Gly Cys He Asp Ala Arg Thr Gly Arg Glu Thr Wing Wing 115 120 125 etc aac tec gtg cgg ctg etc cag age ctg ctg gtg ctc cgc cgc cgg 597 Leu Asn Ser Val Arg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg Arg 130 135 140 ecc tgc tec cgc gac ggc teg ggg etc ecc here ect ggg gcc ttt gcc 645 Pro Cys Ser Arg Asp Gly Ser Gly Leu Pro Thr Pro Gly Ala Phe Ala 145 150 155 160 ttc falls acc gag ttc ate falls gtc ecc gtc ggc tgc acc tgc gtg ctg 693 His Phe Phe Thr Glu Pro Val Val He His Cys Thr Gly Val Leu Cys 165 170 175 cgt tea ecc GTG tgaccgccga ggccgtgggg cccctagact ggacacgtgt Arg Ser Val Pro 745 iso gctccccaga gggcaccccc tatttatgtg tatttattg 805 g tatttatatg cctcccccaa cactaccctt ggggtetggg cattccccgt cagcccccca gtctggagga ctgttctcct 865 catctccagc ctcagtagtt gggggtagaa ggagetcage acctcttcca gcccttaaag 925 ctgcagaaaa ggtgtcacac ggctgcctgt accttggctc cctgtcctgc tcccggcttc 985 ccttacccta tcactggcct caggcccccg caggctgcct cttcccaacc tccttggaag 1045 tacccctgtt tettaaacaa ttatttaagt gtacgtgtat tattaaactg atgaacacat ce 1105 1107 < 210 > 23 < 211 > 197 < 212 > PRT < 213 > primate < 400 > 23 Met Thr Leu Leu Pro Gly Leu Leu Phe Leu Thr Trp Leu His Thr Cys -15 -10 -5 Leu Wing His His Asp Pro Ser Leu Arg Gly His Pro His Ser His Gly -1 1 5 10 15 Thr Pro His Cys Tyr Ser Wing Glu Glu Leu Pro Leu Gly Gln Wing Pro 20 25 30 Pro His Leu Leu Wing Arg Gly Wing Lys Trp Gly Gln Wing Leu Pro Val 40 40 45 Wing Leu Val Ser Ser Leu Glu Wing Wing His Arg Gly Arg His Glu 50 55 60 Arg Pro Be Wing Thr Thr Gln Cys Pro Val Leu Arg Pro Glu Glu Val 65 70 75 Leu Glu Wing Asp Thr His Gln Arg Ser He Ser Pro Trp Arg Tyr Arg 80 85 90 95 Val Asp Thr Asp Glu Asp Arg Tyr Pro Gln Lys Leu Wing Phe Wing Glu 100 105 110 Cys Leu Cys Arg Gly Cys He Asp Wing Arg Thr Gly Arg Glu Thr Wing 115 120 125 Wing Leu Asn Ser Val Arg Leu Leu Gln Ser Leu Leu Val Leu Arg Arg 130 135 140 Arg Pro Cys Ser Arg Asp Gly Ser Gly Leu Pro Thr Pro Gly Wing Phe 145 150 155 Wing Phe His Thr Glu Phe He His Val Val Val Cly Thr Cys Val 160 165 170 175 Leu Pro Arg Ser Val 180 < 210 > 24 < 211 > 403 < 212 > DNA < 213 > primate < 220 > < 221 > feature mis_ < 222 > (1) .. (403) < 223 > note = "n can be a, c, g, or t" < 400 > 24 ttcctgcaca gagaaagagc ccagcgcaac aagtaagcca agaccctgca atgacagtga 60 tggcccagcc atggtcaagt acttgctgct gtcgatattg gggcttgcct ttctgagtga 120 ggcggcagct cggaaaatcc ccaaagtagg acatactttt ttccaaaagc ctgagagttg 180 cccgcctgtg ccaggaggta gtatgaagct tgacattggc atcatcaatg aaaaccagcg 240 cgtttccatg tcacgtaaca tcgagagccg ctccacctcc ccctggaatt acactgtcac 300 ttgggacccc aaccggtacc cctcgaagtt gtacaggccc aagtgtagga aettgggetg tatcaatget 360 403 gtc caaggaaagg aagacatctn catgaattec < 210 > 25 < 211 > 403 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (71) .. (403) < 220 > < 221 > peptide mat_ < 222 > (131) .. (403) < 220 > < 221 > feature mis_ < 222 > (1) .. (403) < 223 > note = "n can be a, c, g, or t; the translation amino acid depends on the genetic code" < 400 > 25 gagaaagagc ttcctgcaca aagtaagcca ccagcgcaac atgacagtga agaccctgca 60 tggcccagcc atg gtc aag tac ttg ctg ctg teg atag tg ggg ett gcc 109 Met Val Lys Tyr Leu Leu Leu Ser He Leu Gly Leu Ala -20 -15 -10 ttt ctg agt gag gcg gca gct cgg aaa ate ecc aaa gta gga cat act 157 Phe Leu Ser Glu Ala Wing Wing Arg Lys He Pro Lys Val Gly His Thr -5 -1 1 5 ttt ttc caa aag ect gag agt tgc ceg ect gtg cea gga ggt agt atg 205 Phe Phe Gln Lys Pro Glu Ser Cys Pro Pro Val Pro Gly Gly Ser Met 10 15 20 25 aag ett gac att ggc ate ate aat gaa aac cag cgc gtt tec atg tea 253 Lys Leu Asp He Gly He He Asn Glu Asn Gln Arg Val Ser Met Ser 30 35 40 cgt aac ate gag age ccc tec ac tc cc tc tc tc tc tc tc tc act act 301 Arg Asn He Glu Ser Arg Ser Thr Ser Pro Trp Asn Tyr Thr Val Thr 45 50 55 tgg gac ecc aac cgg tac ecc teg aag ttg tac agg ecc aag tgt agg 349 Trp Asp Pro Asn Arg Tyr Pro Ser Lys Leu Tyr Arg Pro Lys Cys Arg 60 65 70 aac ttg ggc tgt ate aat gct ca gga aag gaa ate tnc atg aat 397 Asn Leu Gly Cys He Asn Ala Gln Gly Lys Glu Asp He Xaa Met Asn 75 80 85 tec gtc 403 Ser Val 90 < 210 > 26 < 211 > 111 < 212 > PRT < 213 > primate < 400 > 26 Met Val Lys Tyr Leu Leu Leu Ser He Leu Gly Leu Ala Phe Leu Ser -20 -15 -10 -5 Glu Ala Ala Ala Arg Lys He Pro Lys Val Gly His Thr Phe Phe Gln -1 1 5 10 Lys Pro Glu Ser Cys Pro Pro Val Pro Gly Gly Ser Met Lys Leu Asp 15 20 25 He Gly He He Asn Glu Asn Gln Arg Val Ser Met Ser Arg Asn He 30 35 40 Glu Ser Arg Ser Thr Ser Pro Trp Asn Tyr Thr Val Thr Trp Asp Pro 45 50 55 60 Asn Arg Tyr Pro Ser Lys Leu Tyr Arg Pro Lys Cys Arg Asn Leu Gly 65 70 75 Cys He Asn Wing Gln Gly Lys Glu Asp He Xaa Met Asn Ser Val 80 85 90 < 210 > 27 < 211 > 784 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (3) .. (281) < 400 > 27 te gtg ceg tat ett ttt aaa aaa att att ett cae ttt ttt gcc tec 47 Val Pro Tyr Leu Phe Lys Lys He He Leu His Phe Phe Wing Being 1 5 10 15 tat tac ttg tta ggg aga ecc aat ggt agt ttt att ect tgg gga tac 95 Tyr Tyr Leu Leu Gly Arg Pro Asn Gly Ser Phe He Pro Trp Gly Tyr 20 25 30 ata gta aat act tea tta aag teg agt here gaa ttt gat gaa aag tgt 143 He Val Asn Thr Ser Leu Lys Ser Ser Thr Glu Phe Asp Glu Lys Cys 35 40 45 gga tgt gtg gga tgt act gcc gcc ttc aga agt cea falls act gcc tgg 191 Gly Cys Val Gly Cys Thr Wing Wing Phe Arg Ser Pro His Thr Wing Trp 50 55 60 agg gag aga act gct tat tea ctg att aag cat ttg ctg tgt acc 239 Arg Glu Arg Thr Ala Val Tyr Ser Leu He Lys His Leu Leu Cys Thr 65 70 75 aac tac ttt tea tgt et t ate tta att etc ata ata gtc att 281 Asn Tyr Phe Ser Cys Leu He Leu He Leu He Thr Val I 80 85 90 aaaaacccca tgatatttta gaaatctgag aaagagataa agtggtttge tcaaggttat 341 agaacagact accatgtgtt gtatttcaga ttttaattca tgtttgtctg attttaagtt 401 ttgttcgctt gccagggtac cccacaaaaa tgccaggcag ggcattttca tgatgcactt 461 gagatacctg aaatgacagg gtagcatcac aectgagagg ggtaaaggat gggaacctac 521 cttccatggc cgctgcttgg cagtctcttg ctgeatgeta gcagagccac tgtatatgtg 581 ccgaggctct gagaattaac tgcttaaaga actgccttct ggagggagaa gagcacaaga 641 tcacaattaa ccatatacac atcttactgt gcgaggtcat tgagcaatac aggagggatt 701 ttatacattt tageaactat cttcaaaacc tgagctatag ttgtattctg cccccttcct 761 ctgggcaaaa gtgtaaaagt ttg 784 < 210 > 28 < 211 > 93 < 212 > PRT < 213 > primate < 400 > 28 Val Pro Tyr Leu Phe Lys Lys He He Leu His Phe Phe Wing Ser Tyr 1 5 10 15 Tyr Leu Glu Arg Pro Asn Gly Ser Phe He Pro Trp Gly Tyr He 20 25 30 Val Asn Thr Ser Leu Lys Ser Ser Thr Glu Phe Asp Glu Lys Cys Gly 35 40 45 Cys Val Gly Cys Thr Wing Wing Phe Arg Ser Pro His Thr Wing Trp Arg 50 55 60 Glu Arg Thr Wing Val Tyr Ser Leu He Lys His Leu Leu Cys Thr Asn 65 70 75 80 Tyr Phe Ser Cys Leu He Leu He Leu He Thr Val He 85 90 < 210 > 29 < 211 > 460 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (1) .. (189) < 400 > 29 gtg act gta ttg tga gga cag gaa gca caa att ecc atg tgg ate act 48 Val Thr Val Leu Trp Gly Gln Gllu Ala Gln He Pro Met Trp He Thr 1 5 10 15 agg aga gat aat agg tgg ggt cat ttc acc ect tgg tec ect gct tec 96 Arg Arg Asp Asn Lys Trp Gly His Phe Thr Pro Trp Ser Pro Wing Ser 20 25 30 aga ecc aaa gag gcc tac atg gca ttg tgc ttc ett ett agt tgt agg 144 Arg Pro Lys Glu Ala Tyr Met Ala Leu Cys Phe Leu Leu Ser Cys Arg 35 40 45 agg tgt gag ata caa tea ttt gcc tet gac ttt gag ggt tgg tec 189 Arg Cys Glu He Gln Ser Phe Wing Being Asp Phe Glu Gly Trp Ser 50 55 60 tagcatgccc ctgaccagta gccccttaaa tacttcattg atatggaagg tctctgaatc 249 ttcgtgggct taatctacca ctctctgaag ttcttatgtc tttcaaaggc ctctaaaatc 309 tctgccatgt cttgctcatc cagttgttag catgatgtca ttgatacagt ggaetttgga 369 atctaagtgg ggagacactg gtaagtgacc aattacttca cctgtggtgt gcaagecaga 429 tcaggaagcc tctacctgca cgacaacaca t 460 < 210 > 30 < 211 > 63 < 212 > PRT < 213 > primate < 400 > 30 Val Thr Val Leu Trp Gly Gln Glu Wing Gln He Pro Met Trp He Thr 1 5 10 15 Arg Arg Asp Asn Lys Trp Gly His Phe Thr Pro Trp Ser Pro Wing Ser 20 25 30 Arg Pro Lys Glu Wing Tyr Met Wing Leu Cys Phe Leu Leu Ser Cys Arg 35 40 45 Arg Cys Glu He Gln Ser Phe Wing Being Asp Phe Glu Gly Trp Ser 50 55 60 < 210 > 31 < 211 > 150 < 212 > PRT < 213 > rodent < 400 > 31 Met Cys Leu Met Leu Leu Leu Leu Leu Asn Leu Glu Ala Thr Val Lys 1 5 10 15 Wing Wing Val Leu He Pro Gln Ser Val Cys Pro Asn Wing Glu Wing 20 25 30 Asn Asn Phe Leu Gln Asn Val Lys Val Asn Leu Lys Val He Asn Ser 35 40 45 Leu Ser Ser Lys Wing Ser Ser Arg Arg Pro Ser Asp Tyr Leu Asn Arg 50 55 60 Ser Thr Ser Pro Trp Thr Leu Ser Arg Asn Glu Asp Pro Asp Arg Tyr 65 70 75 80 Pro Ser Val He Trp Glu Wing Gln Cys Arg His Gln Arg Cys Val Asn 85 90 95 Wing Glu Gly Lys Leu Asp His His Met Asn Ser Val Leu He Gln Gln 100 105 110 Glu He Leu Val Leu Lys Arg Glu Pro Glu Lys Cys Pro Phe Thr Phe 115 120 125 Arg Val Glu Lys Met Leu Val Gly Val Gly Cys Thr Cys Val Ser Ser 130 135 140 He Val Arg His Wing Ser 145 150 < 210 > 32 < 211 > 147 < 212 > PRT < 213 > rodent < 400 > 32 Met Leu Leu Leu Leu Leu Ser Leu Ala Ala Thr Val Lys Ala Ala Ala 1 5 10 15 He He Pro Gln Ser Be Wing Cys Pro Asn Thr Glu Wing Lys Asp Phe 20 25 30 Leu Gln Asn Val Lys Val Asn Leu Lys Val Phe Asn Ser Leu Gly Wing 35 40 45 Lys Val Ser Ser Arg Arg Pro Ser Asp Tyr Leu Asn Arg Ser Thr Ser 50 55 60 Pro Trp Thr Leu His Arg Asn Glu Asp Pro Asp Arg Tyr Pro Ser Val 65 70 75 80 He Trp Glu Wing Gln Cys Arg His Gln Arg Cys Val Asn Wing Glu Gly 85 90 95 Lys Leu Asp His His Met Asn Ser Val Leu He Gln Gln Glu He Leu 100 105 110 Val Leu Lys Arg Glu Pro Glu Ser Cys Pro Phe Thr Phe Arg Val Glu 115 120 125 Lys Met Leu Val Gly Val Gly Cys Thr Cys Val Ala Ser He Val Arg 130 135 140 Gln Ala Ala 145 < 210 > 33 < 211 > 155 < 212 > PRT < 213 > primate < 400 > 33 Met Thr Pro Gly Lys Thr Ser Leu Val Ser Leu Leu Leu Leu Leu Ser 1 5 10 15 Leu Glu Wing He Val Lys Wing Gly He Thr He Pro Arg Asn Pro Gly 20 25 30 Cys Pro Asn Ser Glu Asp Lys Asn Phe Pro Arg Thr Val Met Val Asn 35 40 45 Leu Asn He His Asn Arg Asn Thr Asn Thr Asn Pro Lys Arg Ser Ser 50 55 60 Asp Tyr Tyr Asn Arg Ser Thr Ser Pro Trp Asn Leu His Arg Asn Glu 65 70 75 80 Asp Pro Glu Arg Tyr Pro Ser Val He Trp Glu Wing Lys Cys Arg His 85 90 95 Leu Gly Cys He Asn Wing Asp Gly Asn Val Asp Tyr His Met Asn Ser 100 105 110 Val Pro He Gln Gln Glu He Leu Val Leu Arg Glu Pro Pro His 115 120 125 Cys Pro Asn Ser Phe Arg Leu Glu Lys He Leu Val Ser Val Gly Cys 130 135 140 Thr Cys Val Thr Pro He Val His His Val Wing 145 150 155 < 210 > 34 < 211 > 151 < 212 > PRT < 213 > viral < 400 > 34 Met Thr Phe Arg Lys Thr Ser Leu Val Leu Leu Leu Leu Leu Ser He 1 5 10 15 Asp Cys He Val Lys Ser Glu He Thr Ser Wing Gln Thr Pro Arg Cys 20 25 30 Leu Wing Wing Asn Asn Being Phe Pro Arg Ser Val Met Val Thr Leu Ser 35 40 45 He Arg Asn Trp Asn Thr Ser Ser Lys Arg Ala Ser Asp Tyr Tyr Asn 50 55 60 Arg Ser Thr Ser Pro Trp Thr Leu His Arg Asn Glu Asp Gln Asp Arg 65 70 75 80 Tyr Pro Ser Val He Trp Glu Wing Lys Cys Arg Tyr Leu Gly Cys Val 85 90 95 Asn Wing Asp Gly Asn Val Asp Tyr His Met Asn Ser Val Pro He Gln 100 105 110 Gln Glu He Leu Val Val Arg Lys Gly His Gln Pro Cys Pro Asn Ser 115 120 125 Phe Arg Leu Glu Lys Met Leu Val Thr Val Gly Cys Thr Cys Val Thr 130 135 140 Pro He Val His Asn Val Asp 145 150

Claims

NOVELTY OF THE INVENTION CLAIMS

1. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising a sequence selected from the group consisting of: a) a mammalian IL-173 sequence that: i) encodes at least a) 147 contiguous amino acids of a coding portion of SEQ ID mature NO: 6; b) 154 contiguous amino acids of a coding portion of mature SEQ ID NO: 8; c) 8 contiguous amino acids of a coding portion of mature SEQ ID NO: 12 or d) two different segments of at least 5 contiguous amino acids of a coding portion of mature SEQ ID NO: 12; ii) encodes the coding portion of mature SEQ ID NO: 6, 8, or 12; (iii) comprises at least a) 441 contiguous nucleotides of the coding portion of mature SEQ ID NO: 5, b) 461 contiguous nucleotides of the coding portion of mature SEQ ID NO: 7 or c) 21 contiguous nucleotides of the coding portion of SEQ ID mature NO: 11; V) comprises the coding portion of mature SEQ ID NO: 5, 7, or 11, or v) comprises the coding portion of mature SEQ ID NO: 9 wherein the polynucleotide is operably linked to a genetic control element; and b) a mammalian IL-174 sequence that: i) encodes at least a) 16 contiguous amino acids of a coding portion of mature SEQ ID NO: 14, b) 140 contiguous amino acids of a coding portion of mature SEQ ID NO: 16, oc ) 31 contiguous amino acids of a coding portion of mature SEQ ID NO: 18; ii) encodes the coding portion of mature SEQ ID NO: 14, 16, or 18; iii) comprises at least a) 27 contiguous nucleotides of the coding portion of mature SEQ ID NO: 13, b) 419 contiguous nucleotides of the coding portion of mature SEQ ID NO: 15 or c) 84 contiguous nucleotides of the coding portion of SEQ Mature ID NO: 17, or iv) comprises the coding portion of mature SEQ ID NO: 13, 15 or 17.

2 - An expression vector, comprising the polynucleotide according to claim 1.

3.- A method for making : a) a polypeptide, comprising expressing said expression vector of claim 2, thereby producing said polypeptide; b) a double nucleic acid, comprising contacting a polynucleotide of claim 2 with a complementary nucleic acid, thereby resulting in the production of said double nucleic acid; or c) a polynucleotide of claim 2, which comprises amplifying using a PCR method.

4. A cell containing said expression vector of claim 2, wherein said cell is: a) a prokaryotic cell; b) a eukaryotic cell; c) a bacterial cell; d) a yeast cell; e) an insect cell; f) a mammalian cell; g) a mouse cell; h) a primate cell; or i) a human cell.

5. An isolated or recombinant antigenic polypeptide comprising a sequence selected from the group consisting of a) (IL-173) comprising at least: i) 147 contiguous amino acids of a coding portion of the mature SEQ ID NO: 6 I) 154 contiguous amino acids of a coding portion of the mature SEQ ID NO: 8, iii) 8 contiguous amino acids of a coding portion of the mature SEQ ID NO: 12, v) two different segments of at least 5 contiguous amino acids of a coding portion of the mature SEQ ID NO: 12; or v) the coding portion of the mature SEQ ID NO: 6, 8, or 12; and b) (IL-174) comprises at least: i) 16 contiguous amino acids of a coding portion of the mature SEQ ID NO: 14 ii) 140 contiguous amino acids of a coding portion of the mature SEQ ID NO: 16, iii) 31 amino acids contiguous of a coding portion of the mature SEQ ID NO: 18; or iv) the coding portion of the mature SEQ ID NO: 14, 16, 0 18.

6. The polypeptide according to claim 5, further characterized in that: a) (IL-173) the polypeptide: i) binds with selectivity to a polyclonal antibody generated against an immunogen derived from the coding portion of the mature SEQ ID NO: 6, 10, or 12; I) is a natural allelic variant of the coding portion of the mature SEQ ID NO: 6, 10, or 12; or iii) has at least two non-overlapping epitopes that are selective for the coding portion of the mature SEQ ID NO: 6, 10, or 12; or b) (IL-174), the polypeptide: i) binds selectively to a polygonal antibody generated against an immunogen derived from the coding portion of the mature SEQ ID NO: 14, 16, or 18; ii) is a natural allelic variant of the coding portion of the mature SEQ ID NO: 14, 16, or 18; or iii) has at least two non-overlapping epitopes that are selective from the coding portion of the mature SEQ ID NO: 14, 16, or 18.

7. The polypeptide of claim 11, which: a) is in a sterile composition; b) it is not glycosylated; c) is denatured; d) is a synthetic polypeptide; e) is attached to a solid substrate; f) is a fusion protein with a detection or purification tag; g) is a substitution of 1 to 5 folds of a natural sequence; or h) is a deletion variant or insertion of a natural sequence.

8. A method for the use of said polypeptide of claim 5: a) for labeling said polypeptide, comprising labeling said polypeptide with a radioactive label; b) to separate said polypeptide from another polypeptide in a mixture, which comprises running the mixture in a chromatography matrix, thus separating said polypeptides; c) to identify a compound that selectively binds said polypeptide, which comprises incubating said compound with said polypeptide under appropriate conditions; thereby causing the compound to bind to said polypeptide; or d) to conjugate said polypeptide to a matrix, which comprises derivatizing said polypeptide with a reacting reagent, and conjugating said polypeptide to said matrix.

9. A binding compound comprising an antigen binding portion of an antibody that binds selectivity to said polypeptide of claim 6, wherein said polypeptide: a) (IL-173) comprises coding portion of SEQ. mature NO: 6, 8, 10, or 12; or b) (IL-174) comprises the coding portion of mature SEQ ID NO: 14, 16, or 18.

10. The binding compound according to claim 9, wherein the antibody is a polyclonal antibody that is raised against: a) (IL-173) the coding portion of the mature SE ID NO: 6, 8, 10, or 12; or b) (IL-174) the coding portion of mature SE ID NO: 14, 16, 0 18.

11. The binding compound of claim 14, wherein said: a) the antibody: i) is immunoselected; ii) binds to a denatured protein; or iii) exhibits a Kd to said polypeptide of at least 30 mM; or b) said linking compound: i) is attached to a solid substrate, including a bead or plastic membrane; ii) is in a sterile composition; or iii) is detectably labeled, including a radioactive or fluorescent label.

12. A method for the production of an antigen: antibody complex, comprising contacting a polypeptide derived from the coding portion of mature SEQ ID NO: 6, 8, 10, 12, 14, 16, or 18, with a linking compound of claim 9, under conditions that allow said complex to be formed.

13. The method of claim 12, wherein said binding compound is an antibody, and said polypeptide is in a biological sample.

14. - A kit comprising said binding compound of claim 19 and: a) a polypeptide of the coding portion of mature SEQ ID NO: 6, 8, 10, 12, 14, 16, or 18 b) instructions for use of said binding compound for detection; or c) instructions for the disposal of said binding compound or other reagents of said equipment.

15. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of mature SEQ ID NO: 5.

16. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of the mature SEQ ID NO: 7.

17. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of the mature SEQ ID NO: 9 wherein the polynucleotide is operably linked to a genetic control element.

18. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of mature SEQ ID NO: 11.

19. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of the mature SEQ ID NO: 13

20. - An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of mature SEQ ID NO: 15.

21. An isolated or recombinant polynucleotide encoding an antigenic polypeptide comprising the coding portion of mature SEQ ID NO: 17.

22. A substantially pure or asylated polypeptide comprising the coding portion of the mature SEQ ID NO: 6.

23. A substantially pure or isolated polypeptide comprising the coding portion of the mature SEQ ID NO: 8. 24. - A substantially pure or asylated polypeptide comprising the coding portion of the mature SEQ ID NO: 10. - A substantially pure or asylated polypeptide comprising the coding portion of the mature SEQ ID NO: 12. 26.- A polypeptide substantially pure or asylated comprising the coding portion of mature SEQ ID NO: 14. 27.- A substantially pure or asylated polypeptide comprising the coding portion of mature SEQ ID NO: 16. 28.- A substantially pure or isolated polypeptide comprising the coding portion of mature SEQ ID NO: 18.