MXPA99009734A - Mammalian cytokine-like factor 7 - Google Patents

Abstract

Novel mammalian zcyto7 polypeptides, polynucleotides encoding the polypeptides, and related compositions and methods including antibodies and anti-idiotypic antibodies.

Description

FACTOR 7 TYPE CTT05INA MAMMERY BACKGROUND OF THE INVENTION The proliferation and differentiation of cells and multicellular organisms are controlled by hormones and polypeptide growth factors. These propagatable molecules allow cells to communicate with each other and act in harmony to form cells and organs, and to repair and regenerate damaged tissue. Examples of hormones and growth factors include steroid hormones (eg, estrogen, testosterone), parathyroid hormone, follicle-stimulating hormone, interleukins, platelet-derived growth factor (PDGF), growth factors epidermal (EGF), the stimulation factor of the granulocyte-macrophage colony (GM-CSF), erythropoietin (EPO) and calcitonin. Hormones and growth factors have an influence on cellular metabolism by binding to proteins. The proteins can be integral membrane proteins that bind to the signaling pathways within the cell, such as REF .: 31880 second messenger systems. Another class of proteins are soluble molecules, such as transcription factors. Of particular interest are cytokines, molecules that promote the proliferation and / or differentiation of cells. Examples of cytokines include erythropoietin (EPO), which stimulates the development of red blood cells; thrombopoietin (TPO), which stimulates the development of cells of the megakaryocyte lineage; and the stimulation factor of the granulocyte colony (G-CSF), which stimulates the development of neutrophils. These cytokines are useful in restoring normal levels of blood cells in patients suffering from anemia or receiving chemotherapy for cancer. The demonstrated in vivo activities of these cytokines illustrate the enormous clinical potential of, and the need for, other cytokines, cytokine agonists and cytokine antagonists.

BRIEF DESCRIPTION OF THE INVENTION The present invention addresses this need by providing a new polypeptide called cytokine type factor 7, hereinafter referred to as Zcyto7 and related compositions and related methods. Thus, one aspect of the present invention provides an isolated Zcyto7 polypeptide having the amino acid sequences as follows. Both human and mouse cDNAs have been discovered. Human sequences are defined by SEQ ID Nos. 2. The murine amino acid and nucleotide sequences are defined by SEQ ID Nos. 11 and 12. The nucleotide sequence of SEQ ID No. 1 contains a reading frame. open coding for a polypeptide of about 180 amino acids in the initial Met as shown in SEQ ID No. 1 and SEQ ID No. 2. A predicted signal sequence is comprised of amino acid residues 1-20, and the Zcyto7 polypeptide mature, predicted, resulting, is represented by the amino acid sequence extending from the amino acid residue 21, a glutamine to and including the amino acid residue 180 a phenylalanine, also represented by SEQ ID No. 14. The correlation data peptides indicate that the mature Zcyto7 may be comprised of a number of mature N-terminal variants including the amino acid sequence extending from the amino acid residue 23, an arginine a and including the amino acid residue 180 of SEQ ID No. 2, also defined by SEQ ID No. 36; the amino acid sequence extending from amino acid residue 27, a serine a and including amino acid residue 180 of SEQ ID No. 2, also defined by SEQ ID No. 37; the amino acid sequence defined by amino acid residue 30, a lysine a and including amino acid residue 180 of SEQ ID No. 2, also defined by SEQ ID No. 38. The amino acid sequence extending from the amino acid 28, a lysine, ay including amino acid residue 180 of SEQ ID No. 2, also defined by SEQ ID No. 41 and the amino acid sequence extending from amino acid residue 53, a methionine, a and including the residue of amino acid 180, also defined by SEQ ID No. 42. The only cleavage observed at the carboxyl terminus is phenylalanine at position 180 which can be completely cleaved. This can occur in all mature Zcyto7 polypeptides, defined above, an example of which is shown by SEQ ID No. 43. Additional variants of human Zcyto7 are defined by SEQ ID Nos. 15-25. Within a further embodiment, the polypeptide further comprises an affinity tag. SEQ ID Nos. 11 and 12 define murine Zcyto7 wherein the mature protein extends between the amino acid residues, amino acid residue 21, a histidine, a and which includes amino acid residue 180, a phenylalanine, also defined by SEQ ID. No. 39; or as an alternative splice site from amino acid residue 23, an arginine, a and including amino acid 180 also defined by SEQ ID No. 40. The present invention is also comprised of polypeptides having an amino acid sequence of at least 90 % identical, more preferably 95%, 97% or 99% identical to those Zcyto7 polypeptides defined above. A further embodiment of the present invention relates to a peptide or polypeptide having the amino acid sequence of a portion having the epitope of a Zcyto7 polypeptide having an amino acid sequence described above. Peptides or polypeptides that have the amino acid sequence of a portion having the epitope of a Zcyto7 polypeptide of the present invention include portions of these polypeptides with at least 9, preferably at least 15 and more preferably at least 30 to 50 amino acids, although polypeptides having epitope of any length up to and which include the complete amino acid sequence of a polypeptide of the present invention described above are also included in the present invention. Examples of the polypeptides are defined by the amino acid sequences of SEQ ID Nos .: 25-35. Also claimed are any of these polypeptides that are fused to another polypeptide or carrier molecule. The present invention further comprises a polypeptide defined by SEQ ID Nos. 15-25, wherein the amino termini of the polypeptides are modified and start at either the amino acid residue 3, an arginine; amino acid residue 7, a serine; the amino acid residue 8, a lysine, the amino acid residue 10, a lysine or the amino acid residue 33, methionine. The present invention is further comprised of a polypeptide wherein the polypeptide is a polypeptide defined by SEQ ID.

Nos. 2, 12, 14-25 and 36 to 42, wherein the amino acid sequences terminate in the isoleucines at amino acid residue 179 of SEQ ID No. 2, at amino acid residue 159 of SEQ ID Nos. 14 -25, which correspond to the amino acid residue 157 of the? EQ ID No. 36, the residue of amino acid 153 of SEQ ID No. 37, the amino acid residue 150 of SEQ ID No. 38, the residue of amino acid 159 of SEQ ID No. 39, amino acid residue 157 of SEQ ID No. 150, amino acid residue 152 of SEQ ID No. 42, amino acid residue 127 of SEQ ID No. 42. The present invention is further comprised of a peptide or polypeptide isolated from the peptides or polypeptides described above having an amino acid sequence modified by the addition, deletion and / or replacement of one or more amino acid residues and maintaining the biological activity of the peptide or polypeptide . Within a further aspect of the invention, there is provided a chimeric polypeptide consisting essentially of a first portion and a second portion bound by a peptide bond. The first portion of the chimeric polypeptide consists essentially of (a) a polypeptide of Zcyto7 as described above (b) allelic variants of the polypeptides described above. The second portion of the chimeric polypeptide consists essentially of another polypeptide such as an affinity tag. Within one embodiment, the affinity tag is an immunoglobulin Fc polypeptide. The invention also provides expression vectors that encode the chimeric polypeptides and transfected host cells to produce the chimeric polypeptides. Another aspect of the present invention provides isolated nucleic acid molecules comprising a polypeptide selected from the group consisting of: (a) a nucleotide sequence encoding the Zcyto7 polypeptides described above; (b) a nucleotide sequence encoding the polypeptides of SEQ ID Nos. 14-40 and (c) a nucleotide sequence complementary to any one of any of the nucleotide sequences in (a) or (b). Additional embodiments of the invention include isolated nucleic acid molecules comprising a polynucleotide having at least 90% identical nucleotide sequence, in more preferred 95%, 97%, 98%, 99% identical to any of the nucleotide sequences in (a), (b) or (c) above, or a polynucleotide that hybridizes under severe hybridization conditions to a polynucleotide having a nucleotide sequence of (a), (b) or (c) above. An additional embodiment of nucleic acid of the present invention relates to an isolated nucleic acid molecule comprising an amino acid of a portion having the epitope of a Zcyto7 polypeptide. Within another aspect of the invention, there is provided an expression vector comprising (a) a transcription promoter; (b) a DNA segment encoding a polypeptide described above, and (c) a transcription terminator, wherein the promoter, the DNA segment and the terminator are linked in operable form. Within a third aspect of the invention, there is provided a cultured eukaryotic cell in which an expression vector has been introduced as described above, wherein the cell expresses a protein polypeptide encoded by the DNA segment. In another embodiment of the present invention is an isolated antibody that binds in a manner specific to a Zcyto7 polypeptide described above. Also claimed is a method for producing antibodies that bind to a Zcyto7 polypeptide comprising inoculating a mammal with a Zcyto7 polypeptide or polypeptide having the epitope in certain Zcyto7, such that the mammal produces antibodies to the polypeptide, and isolating the antibodies. These and other aspects of the invention will become apparent in reference to the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION The teachings of all references cited herein are incorporated herein by reference in their entirety. The term "affinity tag" is used herein to denote a polypeptide segment that can bind to a second polypeptide to provide for the purification or detection of the second polypeptide or to provide the sites for attachment of the second polypeptide to a substrate. In particular, any peptide or protein for which an antibody or other agent is available Specific binding can be used as an affinity tag. Affinity tags include a poly-histidine tract, protein A [Nilsson et al., EMBO J. 4: 1075 (1985); Nilsson et al., Methods Enzymol. 198: 3 (1991)], glutathione S-transferase [Smith and Johnson, Gene 67:31 (1988)], the affinity tag Glu-Glu [Grussen eyer et al., Proc. Nati Acad. Sci. USA 82: 7952-4 (1985)], the substance P, FLAGMR peptide (Hopp et al., Biotechnology 6: 1204-10 (1988), the streptavidin binding peptide, or other antigenic epitope or See, in general, Ford et al., Protein Expression and Purification 2: 95-107 (1991) .The DNAs encoding affinity tags are available from commercial suppliers (eg, Pharmacia Biotech, Piscataway, NJ) The term "allelic variant" denotes any of two or more alternative forms of a gene that occupies the same chromosomal locus.Alice variation occurs naturally through mutation, and can result in phenotypic polymorphisms within the populations.

Gene mutations may be imperceptible (in a change in the encoded polypeptide) or can encode polypeptides having a sequence altered amino acids. The term "allelic variant" is also used herein to denote a protein encoded by an allelic variant of a gene. The term "expression vector" denotes a DNA molecule, linear or circular, comprising a segment encoding a polypeptide of interest operably linked to the additional segments that provide its transcription. These additional segments may include promoter and termite sequences, and may optionally include one or more origins of replication, one or more selectable markers, an enhancer, a polyadenylation signal, and the like. Expression vectors are generally derived from viral or plasmid DNA, or may contain elements of both. The term "isolated" when applied to a polynucleotide molecule, denotes that the polynucleotide has been removed from its natural genetic environment and thus is free of other foreign or unwanted coding sequences, and is in a form suitable for the use within the production systems of genetically engineered proteins. These isolated molecules they are those that separate from their natural environment and include cDNA and genomic clones. The isolated DNA molecules of the present invention are free of other genes with which they are ordinarily associated, but may include 5 'and 3' untranslated regions, which occur naturally such as promoters and terminators. The identification of the associated regions will be apparent to one skilled in the art. See, for example, Dynan and Tijan, Nature 316: 774-78 (1985). When applied to a protein, the term "isolated" indicates that the protein is in a condition different from its native environment, such as part of the animal tissue and blood. In a preferred form, the isolated protein is substantially free of other proteins, particularly other proteins of animal origin. It is preferred to provide the protein in a highly purified form, ie, greater than 95% pure, more preferably greater than 99% pure. The term "operably linked", when referring to DNA segments, denotes that the segments are arranged to function in synchrony for their intended purposes, for example, the transcription starts at the promoter and proceeds through the coding segment to the terminator. The term "polynucleotide" denotes a single or double stranded polymer of deoxyribonucleotide or ribonucleotide bases read from the 5 'end to the 3' end. The polynucleotides include RNA and DNA, and can be isolated from natural sources, synthesized in vitro, or prepared from a combination of natural and synthetic molecules. The term "polynucleotide molecule complements" denotes polynucleotide molecules having a complementary sequence of bases and an inverted orientation as compared to a reference sequence. For example, the sequence 'ATGCACGGG 3' is complementary to 5 'CCCGTGCAT 3'. The term "degenerate nucleotide sequence" denotes a nucleotide sequence that includes one or more degenerate codons (as compared to a reference polynucleotide molecule that codes for a polypeptide). Degenerate codons contain different triplets of nucleotides, but code for the same amino acid residue (ie, the triplets GAU and GAC encode each for Asp).

The term "promoter" denotes a portion of a gene that contains DNA sequences that provide RNA-polymerase binding and initiation of transcription. Promoter sequences are commonly, but not always, found in non-coding regions at 5 'of the genes. The term "secretory signal sequence" denotes a DNA sequence that encodes a polypeptide (a "secretory peptide") that, as a component of a larger polypeptide, directs the larger polypeptide through a secretory pathway of a cell in which it is synthesized. The larger peptide is commonly cleaved to remove the secretory peptide during transit through the secretory pathway. The term "receptor" denotes a protein associated with the cell that binds to a bioactive molecule (ie, a ligand) and measures the effect of the ligand on the cell. Membrane binding receptors are characterized by a multidomain structure comprising an extracellular ligand-binding domain and an intracellular effector domain that is typically comprised in signal transduction. The binding of ligand to the receptor gives This results in a conformational change in the receptor that causes an interaction between the effects domain and other molecules in the cell. This interaction in turn leads to an alteration in the metabolism of the cell. Metabolic events that bind to receptor-ligand interactions include gene transcription, phosphorylation, dephosphorylation, increases in cyclic AMP production, cellular calcium mobilization, membrane lipid mobilization, cell adhesion, lipid hydrolysis of inositol and hydrolysis of phospholipids. Most nuclear receptors also exhibit a multidomain structure, which includes an amino-terminal transacti ation domain, a DNA binding domain and a ligand binding domain. In general, receptors can be bound to the membrane, cytosolic or nuclear; monomeric (eg, thyroid stimulating hormone receptor, beta-adrenergic receptor) or multimeric (eg, PDGF receptor, growth hormone receptor, IL-3 receptor, GM-CSF receptor , G-CSF receptor, erythropoietin receptor and IL-6 receptor).

The term "complement / anti-complement pair" denotes non-identical portions that form a stable pair, associated non-covalently under appropriate conditions. For example, biotin and avidin (or streptavidin) are prototypical members of a complement / anti-complement pair. Other exemplary complement / anti-complement pairs include the receptor / ligand pairs, the antibody / antigen (or hapten or epitope) pairs, the homosentiated / antisense polynucleotide pairs, and the like. Where subsequent dissociation of the complement / anti-complement pair is desirable, the complement / anti-complement pair preferably has a binding affinity of < 109 M1. A "soluble protein" is a protein polypeptide that does not bind to a cell membrane. Within the preferred embodiments of the invention, the isolated polynucleotides hybridize to regions of similar size of the DNA of SEQ ID No. 1, or to a sequence complementary thereto, under severe conditions. In general, severe conditions are selected to be approximately 5 ° C lower than the thermal melting point (Tm) for the specific sequence at an ionic concentration defined and pH. The Tm is the temperature (under a defined ionic concentration and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Typical, severe conditions are those in which the salt concentration is about 0.02 M or less at pH 7 and the temperature is at least about 60 ° C. As noted previously, isolated polynucleotides of the present invention include DNA and RNA. Methods for isolating DNA and RNA are well known in the art. Total RNA can be prepared using guanidine-HCl extraction, followed by isolation by centrifugation in a CsCl gradient [Chirgwin et al., Biochemistry 18: 52-94 (1979)]. Poly (A) + - RNA is prepared from total RNA using the method of Aviv and Leder, Proc. Nati Acad. Sci. USA 69: 1408-1412 (1972). Complementary DNA (cDNA) is prepared from poly (A) + RNA using known methods. The polynucleotides encoding the Zcyto7 polypeptides are then identified and isolated for example by hybridization or PCR. Additionally, the polynucleotides of the present invention can be synthesized using a DNA synthesizer. Currently, the methods of The choice is the phosphoramidite method. If the double-stranded, chemically synthesized DNA is required for an application such as the synthesis of a gene or a gene fragment, then each complementary strand is made separately. The production of short genes (from 60 to 80 bp) is technically direct and can be achieved by synthesizing complementary cells and then fixing them. For the production of longer genes (greater than 300 bp), however, special strategies may be required, because the coupling efficiency of each cycle during chemical synthesis of DNA is rarely 100%. To overcome this problem, synthetic (double-stranded) genes are assembled in modular form from single strand fragments that are 20 to 100 nucleotides in length. In addition to coding the protein, synthetic genes can be designed with terminal sequences that facilitate insertion into a restriction endonuclease site and a cloning vector and other sequences must also be added containing appropriate signals for initiation and termination. the transcription and translation.

See Glic, Bernard R. and Jack J. Pasternak, Molecular Biotechnology, Principies & applications of Recombinant DNA, (ASM Press, Washington, D.C. 1994), Itakura, K. et al., Synthesis and use of synthetic oligonucleotides. Annu. Rev. Biochem. 53: 323-356 (1984), and Cumie, S. et al. chemical synthesis of the thymidylate synthase gene. Proc. Nati Acad. Sci. USA 87: 633-637 (1990). Those skilled in the art will recognize that the sequences described in SEQ ID Nos. 1 and 2 represent an individual allele of a human. There are a number of N-terminal variants, mature, that occur naturally that have the guiding sequence split into different positions. They include the sequences defined by SEQ ID Nos. 14, 36, 37 and 38. Allelic variants of these sequences can be cloned by probing the cDNA or genomic libraries of different individuals according to standard procedures. Examples of variants of human Zcyto7 are represented by the polypeptides of SEQ ID Nos. 15-25. The Zcyto7-murine cDNA and the protein are described by SEQ ID Nos. 11 and 12. The Zcyto7 murine polypeptide is defined by SEQ ID Nos. 39 and 40. The present invention additionally provides counterpart proteins and polynucleotides from other species ("species orthologs"). Of particular interest are the Zcyto7 polypeptides from other mammalian species, including murine, porcine, ovine, bovine, canine, feline, equine, and other primates. The orthologs of Zcyto7 human protein species can be cloned using the information and compositions provided by the present invention in combination with conventional cloning techniques. For example, a cDNA can be cloned using the mRNA obtained from a tissue or cell type that expresses the protein. Suitable sources of mRNA can be identified by probing Northern blots with probes designed from the sequences described herein. Then a library can be prepared from the mRNA and a positive tissue or cell line. A cDNA encoding the protein can then be isolated by a variety of methods, such as by probing with a human or mouse cDNA, complete or partial, or with one or more sets of proteins. degenerate probes traced in the described sequences. A cDNA can also be cloned using the polymerase chain reaction, or PCR, (Mullis, U.S. Patent No. 4,683,202), using primers designed from the sequences described herein. Within a further method, the cDNA library can be used to transform or transfect host cells, and expression of the cDNA of interest can be detected with an antibody to the protein. Similar techniques can also be applied to the isolation of genomic clones. As used and claimed, the text "an isolated polynucleotide encoding a polypeptide, the polynucleotide defined by SEQ ID No. 2" includes all species allelic and orthologous variants of the polypeptides of SEQ ID No. 2. The present invention also provides isolated protein polypeptides that are substantially homologous to the protein polypeptides of SEQ ID No. 2 and their species orthologs. By "isolated" is meant a protein or polypeptide that is in a condition different from its native environment, such as separated from blood and animal tissue. In a way Preferred, the isolated polypeptide is substantially free of other polypeptides, particularly other polypeptides of animal origin. It is preferred to provide the polypeptides in a highly purified form, ie, greater than 95% pure, more preferably more than 99% pure. The term "substantially homologous" is used herein to denote polypeptides having 50%, preferably 60%, more preferably at least 80%, sequence identity to the sequence shown in SEQ ID No. 2, or your species orthologs. These species will be more preferably at least 90% identical, and more preferably 95% or more identical to SEQ ID No. 2, or their species orthologs. The percentage of sequence identity is determined by conventional methods. See, for example, Altschul et al., Bull, Math. Biol. 48: 603-616 (1986) and Heníkoff and Henikoff, Proc. Nati Acad. Sci. USA 89: 10915-10919 (1992). Briefly, two amino acid sequences are aligned to optimize the alignment scores using a separation gap penalty 10, a separation extension penalty of 1, and a "blossom 62" scoring matrix of Henikoff and Henikoff (ibid. ) as shown in Table 2 (amino acids are indicated by the normal code of a letter). The identity percent is then calculated as: Total number of identical matches x 100 [length of the longest sequence plus the number of separations entered in the longest sequence in order to align the two sequences] The sequence identity of the polynucleotide molecules is determined by similar methods using a ratio as described above. Substantially homologous proteins and polypeptides are characterized since they have one or more amino acid substitutions, deletions or additions. These changes are preferably of a minor nature, which are substitutions of conservative amino acids (see Table 3) and other substitutions that do not significantly affect the fold or activity of the protein or polypeptide; small deletions, typically from one to about 30 amino acids; and small amino- or carboxy-terminal extensions, such as the amino-terminal methionine residue, a small binding peptide of up to about 20-25 residues, or a small extension that facilitates purification (an affinity tag), such as a poly-histidine tract, protein A [Nilsson et al., EMBO J. 4: 1075 (1985); Nilsson et al., Methods Enzymol. 198: 3, (1991)], glutathione-S-transferase [Smith and Johnson, Gene 67:31, (1988)], or other binding domain or antigenic epitope. See, in general Ford et al., Protein Expression and Purification 2. 95-107 (1991). The DNA that encode for affinity marks - are available from commercial suppliers (eg Pharmacia Biotech, Piscataway, NJ).

Table 3 Conservative amino acid substitutions The essential amino acids in the polypeptides of the present invention can be identified according to methods known in the art, such as sequence directed mutagenesis or alanine scanning mutagenesis [Cunningham and Wells, Science 244: 1081-1085 ( 1989); Bass et al., Proc. Ntl. Acad. Sci. USA 88: 4498-4502 (1991)]. In this latter technique, individual alanine mutations are introduced into each residue in the molecule, and the resulting mutant molecules are tested for biological activity (e.g., ligand binding and signal transduction) to identify the amino acid residues that are critical for the activity of the molecule. The ligand-protein interaction sites can also be determined by analysis of the crystal structure as determined by these techniques such as nuclear magnetic resonance, crystallography or photoaffinity-labeled. See, for example, de vos et al., Science 255: 306-312 (1992); Smith et al., J. Mol. Biol. 224: 899-904, 1992; Wlodaver et al., FEBS Lett. 309: 59-64 (1992). The identities of the essential amino acids can also be inferred from analysis of homologies with related proteins.

Multiple amino acid substitutions can be made and tested using known methods of mutagenesis and detection, such as those described by Reidhaar-Olson and Sauer, Science 241: 53-57 (1988) or Bowie and Sauer, Proc. Nati Acad. Sci. USA 86: 2152-2156 (1989). Briefly, these authors describe methods for the simultaneous randomization of two or more positions in a polypeptide, selecting the functional polypeptide, and then sequencing the utagenized polypeptides to determine the spectrum of the permissible substitutions in each position. Other methods that include phage display can be used, for example, Lowman et al., Biochem. 30: 10832-10837 (1991); Ladner et al., U.S. Patent No. 5,223,409; Huse, WIPO Publication WO 92/06204) and region-directed mutagenesis, Derbyshire et al., Gene 46: 145 (1986); Ner et al., DNA 7: 127 (1988). Mutagenesis methods as described above can be combined with high production detection methods to detect the activity of the mutagenized proteins, cloned in the host cells. The preferred analyzes in this regard include cell proliferation and ligand binding assays based on a biosector, which are described below. DNA and mutagenized molecules that code for active proteins or portions thereof (e.g., ligand binding fragments) can be recovered from the host cells and sequenced rapidly using modern equipment. These kits allow rapid determination of the importance of individual amino acid residues in a polypeptide of interest, and can be applied to polypeptides of an unknown structure. Using the methods described above, one skilled in the art can prepare a variety of polypeptides that are substantially homologous to SEQ ID No. 2 or allelic variants thereof and retain the properties of the wild-type protein. As expressed and claimed herein, the text "a polypeptide as defined by SEQ ID No. 2" includes all allelic and orthologous species variants of the polypeptide. Another embodiment of the present invention provides a peptide or polypeptide comprising a portion having epitope of a polypeptide of the invention. The epitope of this portion of polypeptide is an immunogenic or antigenic epitope and a polypeptide of the invention. A region of a protein to which an antibody can be linked is defined as an "antigenic epitope". See, for example Geysen, H.M. et al., Proc. Nati Acad Sci. USA 81: 3998-4002 (1984). As for the selection of peptides or polypeptides having an antigenic epitope (ie, containing a region of a protein molecule to which an antibody can be attached), it is well known in the art that relatively short synthetic peptides that they mimic part of a protein sequence are routinely capable of producing an antiserum that reacts with the partially unchanged protein. See Sutcliffe, J.G. et al. Science 219: 660-666 (1983). Peptides capable of producing protein reactive sera are frequently represented in the primary sequence of a protein, can be characterized by a set of simple chemical rules, and are not confined to the immunodominant regions of intact proteins (ie, epitopes) immunogenic) or amino or carboxyl terminals. Peptides that are extremely hydrophobic and those with six or less residues in are generally ineffective in the induction of antibodies that bind to the mimicked protein; Smaller soluble peptides, especially those containing proline residues, are usually effective. The peptides and polypeptides having the epitope, antigenic, of the invention are therefore useful for formulating antibodies, including monoclonal antibodies, which specifically bind to a polypeptide of the invention. Peptides having the epitope, antigenic, and polypeptides of the present invention contain a sequence of at least nine, preferably between 15 to about 30 amino acids contained within the amino acid sequence of a polypeptide of the invention. However, peptides or polypeptides comprising a larger portion of an amino acid sequence of the invention, containing from 30 to 50 amino acids, or any length up to and including the complete amino acid sequence of a polypeptide of the invention, also they are useful for inducing antibodies that react with the protein. Preferably, the amino acid sequence of the peptide having the epitope and selected for provide substantial solubility in aqueous solvents (ie, the sequence includes relatively hydrophilic residues and hydrophobic residues and is preferentially avoided); and sequences containing proline residues are particularly preferred. All polypeptides shown in the sequence listing contain antigenic epitopes to be used according to the present invention, however, the specifically designed antigenic epitopes include the peptides defined by SEQ ID Nos. 27-35. The polynucleotides, generally a cDNA sequence, of the present invention encode the polypeptides described above. A cDNA sequence encoding a polypeptide of the present invention is comprised of a series of codons, each amino acid residue of the polypeptide that is encoded by a codon and each codon that is comprised of three nucleotides. The amino acid residues are encoded by their respective codons as follows. Alanine (Ala) is encoded by GCA, GCC, GCG or GCT; Cysteine (Cys) is encoded by TGC or TGT; Aspartic acid (Asp) is encoded by GAC or GAT; Glutamic Acid (Glu) is encoded by GAA or GAG; 5 Phenylalanine (Phe) is encoded by TTC or TTT; Glycine (GLY) is encoded by GGA, GGC, GGG or GGT; Histidine (His) is encoded by CAC or CAT; Isoleucine (lie) is encoded by ATA, ATC or ATT; Lysine (Lys) is encoded by AAA, or AAG; Leucine (Leu) is encoded by TTA, TTG, CTA, CTC, CTG or CTT; 15 Methionine (Met) is encoded by ATG; Asparagine (Asn) is encoded by AAC or AAT; Proline (Pro) is encoded by CCA, CCC, CCG or CCT; Glutamine (Gln) is encoded by CAA or CAG; 20 Arginine (arg) is encoded by AGA, AGG, CGA, CGC, CGG or CGT; Serine (Ser) is encoded by AGC, AGT, TCA, TCC, TCG or TCT; Threonine (Thr) is encoded by ACA, ACC, 25 ACG or ACT; Valine (Val) is encoded by GTA, GTC, GTG or GTT; Tryptophan (Trp) is encoded by TGG; and Tyrosine (Tyr) is coded by TAC or TAT.

It is to be recognized that according to the present invention, when a cDNA is claimed as described above, it is understood that what is claimed is both the sense strand, the antisense strand, and the DNA as an individual strand that has both the strand homosense and antisense are bound together by their respective hydrogen bonds. Messenger RNA is also claimed (MRNA) encoding the polypeptides of the present invention, and the mRNA that is encoded by the CDNA described above. A messenger RNA (MRNA) will code for a polypeptide using the same codons as those defined above, with the exception that each thymidine nucleotide (T) is replaced by a uracil nucleotide (U). The protein polypeptides of the present invention, which include full-length proteins, protein fragments (e.g., receptor binding fragments) and polypeptides of Pressure can be produced in genetically engineered host cells according to conventional techniques. Suitable host cells are those types of cells that can be transformed or transfected with exogenous DNA and grow the culture, and include bacteria, fungal cells, and higher, cultured eukaryotic cells. Eukaryotic cells, particularly cultured cells of multicellular organisms, are preferred. Techniques for manipulating cloned DNA molecules and for introducing exogenous DNA into a variety of host cells are described by Sambrook et al., Molecular Cloning: A Laboratory Manual, (2nd ed.) (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989), and Ausubel et al., Ibid. In general, a DNA sequence encoding a Zcyto7 polypeptide is operably linked to other genetic elements required for its expression, generally including a promoter and transcription terminator, within an expression vector. The vector will also commonly contain one or more selectable markers and one or more origins of replication, although those skilled in the art will recognize that within certain systems can be provided with selectable markers of separate vectors, and replication of the exogenous DNA can be provided by integration into the host cell genome. The selection of promoters, terminators, selectable markers, vectors and other elements is one of routine design within the level of ordinary skill in the art. Many of these elements are described in the literature and are available through commercial providers. To direct a Zcyto7 polypeptide in the secretory pathway of a host cell, a secretory signal sequence (also known as a leader sequence, prepro sequence or pre-sequence) is provided in the expression vector. The secretory signal sequence may be that of the protein, or it may be derived from another protein secreted (eg, t-PA) or synthesized de novo. The secretory sequence binds to the Zcyto7 DNA sequence in the correct reading frame. The secretory signal sequences are commonly placed 5 'to the DNA sequence encoding the polypeptide of interest, although certain signal sequences can be placed anywhere in the DNA sequence of interest (see, for example, Welch et al., U.S. Patent No. 5,037,743; Holland et al., U.S. Patent No. 5, 143, 830). Cultured mammalian cells are preferred hosts within the present invention. Methods for producing exogenous DNA in mammalian host cells include calcium phosphate mediated transfection, Wigler et al., Cell 14: 725 (1978); Corsaro and Pearson, somatic Cell Genetics 7: 603 (1981): Graham and Van der Eb, virology 52: 456 (1973), electroporation, Neumann et al., EMBO J. 1: 841-845 (1982), transfection mediated by DEAE-dextran, Ausubel et al., Eds., Current Protocols in Molecular Biology (John Wiley and Sons, Inc., NY, 987), and liposome-mediated transfection (Hawley-Nelson et al., Focus 15:73 (1993 ), Ciccarone et al., Focus 15:80 (1993) The production of recombinant polypeptides in cultured mammalian cells is described, for example, by Levinson et al., U.S. Patent No. 4,713,339; Hagen et al. , U.S. Patent No. 4,784,950; Palmiter et al., U.S. Patent No. 4,579,821; and Ringold, U.S. Patent No. 4,656,134.

Cultured mammalian crops include COS-1 (ATTC No. CRL 1650), COS-7 (ATCC No. CRL 1651), BHK (ATCC No. CRL 1632), BHK 570 (ATCC No. CRL 10314), 293 [ATCC No. CRL 1573; Graham et al., J. Gen. Virol. 36: 59-72 (1977)] and Chinese hamster ovary cell lines (e.g. CHO-K1; ATCC No. CCL 61). Additional, suitable cell lines are known in the art and are available from public depositories such as the American type Culture collection, Rockville, Maryland. In general, strong transcription promoters, such as the SV-40 or cytomegalovirus promoters, are preferred. See, for example, U.S. Patent No. 4,956,288. Other suitable promoters include those of the metallomyonein genes (U.S. Patent Nos. 4,579,821 and 4,601,978) and the adenovirus major late promoter. The selection of drugs is generally used to select the cultured mammalian cells into which the foreign DNA has been inserted. These cells are commonly referred to as "transfectants". Cells that have been cultured in the presence of the selective agent and are able to pass the gene of interest to their progeny are they require as "stable transfectants". A preferred selectable marker is a gene that codes for resistance to the antibiotic neomycin. The selection is carried out in the presence of a neomycin-type drug, such as G-418 or the like. Selection systems can also be used to increase the level of expression of the gene of interest, or a process referred to as "amplification". The amplification is carried out by culturing the transfectants in the presence of a low level of the selective agent and then by increasing the amount of the selective agent to select the cells that produce high levels of the introduced gene products. A selectable, amplifiable, preferred marker is dihydrofolate reductase, which confers resistance to methotrexate. Other drug resistance genes (for example, hygromycin resistance, multi-drug resistance, puromycin-acetyltransferase) may also be used. Other higher eukaryotic cells can also be used as hosts, including insect cells, plant cells and poultry cells. The transformation of insect cells and the production of foreign polypeptides in the present is described by Guarino et al., U.S. Patent No. 5,162,222; Bang et al., U.S. Patent No. 4,775,624; and WIPO publication WO 94/06463. The use of Agrobacterium rhizogenes as a vector for expressing genes in plant cells has been reviewed by Sinkar et al., J. Biosci. (Bangalore) 11: 47-58 (1987). Fungal cells, including yeast cells, and particularly cells of the genus Saccharomyces can also be used with the present invention, such as to produce protein fragments or polypeptide fusions. Methods for transforming yeast cells with exogenous DNA and producing recombinant polypeptides therefrom are described, for example, in Kawasaki, U.S. Patent No. 4,599,311; Kawasaki et al., U.S. Patent No. 4,931,373; Brake, U.S. Patent No. 4,870,008; Welch et al., U.S. Patent No. 5,037,743; and Murray et al., U.S. Patent No. 4,845,075. Transformed cells are selected by the phenotype determined by the selectable marker, commonly the drug resistance or the capacity to grow in the absence of a particular nutrient (for example, leucine). A preferred vector system for the use of yeast is the POTl vector system described by Kawasaki et al., U.S. Patent No. 4,931,373, which allows transformed cells to be selected for growth in the glucose-containing medium. Promoters and terminators suitable for the use of the yeast include those from the genes of the glycolytic enzyme (see, for example, Kawasaki, U.S. Patent No. 4,599,311, Kingsman et al., U.S. Pat. No. 4,615,974; and Bitterr, U.S. Patent No. 4,977,092) and the alcohol dehydrogenase genes. See also, U.S. Patent Nos. 4,990,446; 5,063,154; 5,139,936 and 4,661,454. Transformation systems for other yeasts, including Hansenula polymorpha, Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces fragilis, Ustilago mayis, Pichia pastoris, Pichia methanolíca, Pichia guillermondii and Candida maltose are known in the art. See, for example, Gleeson et al., J. Gen. Microbiol. 132: 3459-3465 (1986) and Cregg, U.S. Patent No. 4,882,279. Aspergillus cells can be used from according to the methods of McKnight et al., U.S. Patent No. 4,935,349. Methods for transforming the Acremon i um chrysogen um are described by Sumino et al., U.S. Patent No. 5,162,228. Methods for transforming Neurospora are described by Lambowitz, U.S. Patent No. 4,486,533. Transformed or transfected host cells are cultured according to conventional procedures in a culture medium containing nutrients and other components referred to the growth of the chosen host cells. A variety of suitable means, including defined means and complex media, are known in the art and generally include a source of carbon, a source of nitrogen, essential amino acids, vitamins and minerals. The media may also contain components such as growth factors or serum, as required. The growth medium will generally be selected for cells containing the exogenously added DNA, for example, drug selection or drug deficiency in an essential nutrient that is complemented by the selectable marker carried in the expression vector or co-transfected from the host cell. Within one aspect of the present invention, a new protein is produced by a cultured cell, and the cell is used to detect a receptor or receptors for the protein, including the natural receptor, as well as natural ligand agonists and antagonists.

PROTEIN ISOLATION The expressed recombinant polypeptides (or chimeric polypeptides) can be purified using conventional fractionation and / or purification methods and media for these. Precipitation with ammonium sulfate and extraction with acid or chaotrope can be used for fractionation of the sample. Exemplary purification steps may include inverted phase high performance liquid chromatography and FPLC, size exclusion, and hydroxyapatite. Suitable anionic exchange media include derivatized dextrans, agarose, cellulose, polyacrylamide, specialized silicas and the like. PEI, DEAE, QAE and Q derivatives are preferred, with the DEAE fast-flow safarosa (Pharmacia, Piscataway) that is particularly preferred. Exemplary chromatographic media include those media derivatized with phenyl, butyl, or octyl groups, such as phenyl-separable FF (Pharmacy) Toyopearl butyl 650 (Toso Haas, Montgomeryville, PA), octyl-safarose (Pharmacia) and the like; or polyacrylic resins, such as Amberchrom CG 71 (Toso Haas) and the like. Suitable solid supports include glass beads, silica based resins, cellulosic resins, agarose beads, crosslinked agarose beads, polystyrene beads, crosslinked polyacrylamide resins and the like, which are insoluble under the conditions in which they are to be used . These supports can be modified with reactive groups that allow the binding of the proteins by the amino groups, carboxyl groups, sulfhydryl groups, hydroxyl groups and / or carbohydrate moieties. Examples of coupling chemicals include activation with cyanogen bromide, activation with N-hydroxysuccinimide, activation with epoxide, activation with sulfhydryl, activation with hydrazide, and carboxyl and amino derivatives for the carbodiimide coupling chemicals.

These and other solid media are well known and widely used in the art, and are available from commercial suppliers. Methods for attaching the receptor polypeptides to the support media are well known in the art. The selection of a particular method is a routine matter and is determined in part by the properties of the chosen support. See, for example, Affini and Chroma tography: Principi is & Methods (Pharmacia LKG Biotechnology, Uppsala, Sweden, 1988). The polypeptides of the present invention can be isolated by exploiting their properties. For example, chromatography with immobilized metal ion uptake (IMAC) can be used to purify histidine rich proteins. Briefly, a gel is charged first with divalent metal ions to form a chelate [E. Sulkowski, Trends i n Bi och e. 3: 1-1 (1985)]. The proteins rich in histidine will be absorbed to this matrix with different affinities, depending on the metal ion used and eluted by competitive elution, decreasing the pH of the use of strong chelating agents. Other methods of purification include protein purification glycosylated by lectin affinity chromatography and ion exchange chromatography. [Me th ods i n In zumol. , Vol. 152: 529-39, "Guide to Protein Purification", M. Deutscher, (ed.), (Acad. Press, San Diego, 1990. Alternatively, a fusion of polypeptide of interest and one can be constructed. affinity tag (eg, polystidine, maltose binding protein, an immunoglobulin domain) to facilitate purification Additionally, to facilitate purification of the secreted receptor polypeptide, a carboxyl-terminal extension such as a poly-hist idine tag , P peptide FLAG® [HOPP ET AL., blO / Techn olgy 5: 1204-1210 (1988); available from Eastman Kodak Co., New Haven, Ct, a Glu-Glu affinity tag [Grussenmeyer et al. , Proc. Na ti, Acad. Sci. USA 82: 7952-4 (1985)], or another polypeptide or protein for which an antibody or other specific agent is available, can be fused to Zcyto7 to aid in purification.

Applications Northern blot analysis of Zcyto7 expression revealed that Zcyto7 is expressed specifically in the spinal cord. In situ analysis of the spinal cord reveals that this expression is located in the neurons and dorsal root ganglia. Therefore Z cyt or l can play a role in the maintenance of the spinal cord which comprises either glial cells or neurons. This indicates that Zcyto7 can be used to treat a variety of degenerative diseases such as amyotrophic lateral sclerosis (ALS), or dimellination diseases including multiple sclerosis. Z c yt ol can also be used to treat sensory neuropathies. The tissue specificity of Zcyto7 expression suggests that Zcyto7 may be a growth and / or maintenance factor in the spinal cord. The location of the Zcyto7 gene on chromosome 5 indicates that Zcyto7 is a cytosine that can be used to modulate the activities of immune system cells. Zcyto7 can also be used as a chemoattractant of neutrophils in the spinal column. This will be useful as an anti-infective for infections in the spinal column. It can also be used to help regulate other cytokines in the spinal cord. Zcyto7 can also be given to treat peripheral neuropathies such as Charcot-Maria tooth disease (CMT) which is located in the same chromosomal region of 5q as Zcyto7. The fact that Zcyto inhibits the growth of BAF-3 cells and TF-1 cells as shown in Examples 11 and 12 below indicates that Zcyto7 can be used to treat immune diseases and possibly cancers such as leukemias. The present invention also provides reagents that will find use in diagnostic applications. For example, the Zcyto7 gene is expressed heavily in the spinal cord. An area comprising Zcyto7 DNA or RNA or a subsequence thereof can be used to determine whether Zcyto7 is present on chromosome 5 or whether a mutation has occurred. The present invention also provides reagents with significant therapeutic value. The Zcyto7 polypeptide (which occurs naturally or recombinantly), fragments thereof, antibodies and anti-idiotypic antibody, together with the compounds identified as having affinity and binding to the Zcyto7 polypeptide, should be useful in the treatment and conditions associated with physiology or normal development, including abnormal proliferation, for example, cancerous conditions or degenerative conditions. For example, a disease or disorder associated with abnormal expression or abnormal signaling by a Zcyto7 polypeptide should be a likely target for an agonist or antagonist of the Zcyto7 polypeptide. In particular, Zcyto7 can be used to treat inflammation. Inflammation is the result of an immune response to an infection or as an autoimmune response to a self-antigen. Treatment doses should be titrated to optimize safety and efficiency. Methods for administration include intravenous, peritoneal, intramuscular, subdural, spinal fluid or transdermal administration. The pharmaceutically acceptable carriers will include water, saline, buffers to name a few. Dose ranges will ordinarily be expected from 0.1 ug to 1 mg per kilogram of body weight per day. Preferably from 1 ug to 100 ug per day. However, the doses may be higher or lower, as can be determined by a doctor with skill in the art. For a full discussion of drug formulations or dose ranges see Remington's Pharmaceutical Sciences, 17th Ed., (Mack Publishing Co., Easton, Penn., 1990), and Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 9th Ed. (Pergamon Press 1996).

Use of Zcyto7 to promote growth of bone and cartilage It has been found that Zcyto7 stimulates the proliferation of both chondrocytes and osteoblasts as shown in examples 7 and 9, respectively. In addition, Zcyto7 also stimulates the stable level of glycosaminoglycan present in chondrocyte cultures as shown in example 8. Thus, Zcytol can be used to stimulate the growth of both bone and cartilage in a variety of different therapeutic environments. The Zcyto7 can be implanted in a mammalian body so that the Zcyto7 is in contact with the osteoblasts so that the proliferation of the osteoblasts occurs and the bone growth. For example, Z c and t or l can be placed in a matrix [with or without a bone morphogenic protein (BMP)]. The BMP induces the migration of the mesenchymal precursors of the osteoblasts to the site and further induces the differentiation of the mesenchymal cells in the osteoblast. Z cyt or l will then stimulate further proliferation of osteoblasts. A suitable matrix is made of particles of porous materials. The pores must be of one dimension to allow migration of the progenitor cells and subsequent differentiation and proliferation. An ideal particle size should be in the range of 70-850 mm, preferably 150-420 mm. The matrix containing the Zcyto7 can be molded into a shape that encompasses a bone defect. The effects of matrix materials are bone in the form of demineralized particles, extracted from guanidine, specific to the species. Other potentially useful matrix materials include collagen, homopolymers and copolymers of glycolic acid and lactic acid, hydroxyapatite, tricalcium phosphate and other calcium phosphates. The Zcyto7 can be applied in a matrix at a sufficient concentration to promote the proliferation of osteoblasts, preferably a concentration of at least 1 ug / ml of matrix. A solution of Zcyto7 can also be injected directly into the site of a bone fracture or bone defect that includes areas of bone degeneration to accelerate the healing of the fracture or defect site. Examples of BMPs and the use of matrices to produce are described in PCT application publication No. WO 92/07073, publication No. WO 91/05802, United States Patent No. 5,645,591 and United States patent. United No. 5,108,753. Zcyto7 can be used additionally to treat osteoporosis by administering a therapeutically effective amount of Zcyto7 to an individual. A preferred dose will be 1 ug of Zcyto7 per kilogram of body weight per day. As noted above, it has been determined that Zcyto7 can be used to promote the production of cartilage through its ability to stimulate the proliferation of chondrocytes. Zcyto7 can be injected directly into the site where the cartilage will grow. For example, Zcyto7 can be injected directly into the joints that have been afflicted with the osteoarthritis or other injured sites in which cartilage has been worn. An example of a case in which additional cartilage needs to be grown is the shoulders and knees of injured athletes. You can also grow the cartilage by first removing the chondrocytes from an individual by growing the chondrocytes with Zcyto7 so that they proliferate and reimplant the chondrocytes back to the individual where the cartilage is needed. Zcyto7 can also be used to stimulate the generation of dentin or bone that has been lost due to periodontal disease. To do this, the surrounding tissue must be completely cleaned and a Zcto 7 solution is administered preferably by injection at the site where the regeneration of the dentin is desired. The antibodies to the Zcyto7 polypeptide can be purified and then administered to a patient. These reagents can be combined for therapeutic use with active or inert ingredients, for example, in pharmaceutically acceptable carriers or diluents together with stabilizers and excipients physiologically.

These combinations can be filtered in a sterile manner and can be placed in dosage form as by lyophilization in dose or storage bottles in stabilized aqueous preparations. This invention also contemplates the use of antibodies, binding fragments thereof or single chain antibodies of the antibodies that include forms that are not complement binding. The amounts of reagents necessary for effective therapy will depend on many different factors, including the means of administration, target site, physiological state of the patient, and other medications administered. In this way, treatment doses should be titrated to optimize productivity and effectiveness. Typically, doses used in vitro may provide useful guidance in amounts useful for the in vivo administration of these reagents. The animal test of the effective dose for the treatment of particular disorders will provide additional predictive indication in the human dose. Methods for administration include oral, intravenous, peritoneal, intramuscular, or transdermal administration. The carriers pharmaceutically acceptable will include water, saline, buffers to name but a few. Dose ranges will ordinarily be expected from 1 ug to 1000 ug per kilogram of body weight per day. However, the doses may be higher or lower as may be administered by a physician skilled in the art. For a complete discussion of drug formulations and dose ranges see Remington's Pharmaceutical Sciences, 17th Ed., (Mack Publishing Co., Easton, Penn., 1990), and Goodman and GilmanO's: The Pharmacological Bases of Therapeutics, 9th Ed.

(Pergamon Press 1996).

Therapeutic treatment based on nucleic acids If a mammal has a mutated Zcyto7 gene or lacks it, the Zcyto7 gene can be introduced into the mammalian cells. In one embodiment, a gene encoding a Zcyto7 polypeptide is introduced in vivo into a viral vector. These vectors include DNA attenuated or defective viruses such as, but not limited to, herpes simplex virus (HSV), papillomavirus, Epstein Barr virus (EBV), adenovirus, adeno-associated virus (AAV), and Similar. Defective viruses, which completely or almost completely lack viral genes, are preferred. A defective virus is not ineffective after introduction into a cell. The use of defective viral vectors allows administration to cells in a specific localized area, without the interest that the vector can infect other cells. Example-s of particular vectors include, but are not limited to, herpes defective virus 1 vector (HSV1) [Kaplitt et al., Molec. Cell. Neurosci. , 2: 320-330 (1991)], an attenuated adenovirus vector, such as the vector described by St rat ford-Perricaudet et al., J. Virol., 61: 3096-3101 (1987); [Samulski et al., J. Virol., 53: 3822-3828 (1989).) In another embodiment, the gene can be introduced into a retroviral vector, for example, as described in Anderson et al., United States patent. No. 5,399,346; Mann et al., Cell, 33: 153 (1983); Temin et al., U.S. Patent No. 4,650,764; Temin et al., U.S. Patent No. 4,980,289; Markowitz et al. al., J. Virol., 62: 1120 (1988), Temin et al., U.S. Patent No. 5,124,263, International Patent Publication No. WO 95/07358, published on 16 March 1996 by Dougherty et al .; and Blood, 82: 845 (1983). Alternatively, the vector can be introduced by lipofection in vivo using liposomes. Synthetic cationic lipids can be used to prepare liposomes for the transfection in vivo of a gene encoding a marker [Felgner et al. , Proc. Nati Acad. Sci. USA, 54: 7413-7417 (1987); see Mackey et al. , Proc. Nati Acad. Sci. USA, 55: 8027-8031 (1988)]. The use of lipofection to introduce endogenous genes into specific organs in vivo has certain practical advantages. The molecular search of the objective of liposomes to specific cells represents an area of benefit. It is clear that the direction of transfection to particular cells represents an area of benefit. It is clear that the direction of transfection to particular cell types will be particularly advantageous in a tissue with cellular heterogeneity such as the pancreas, liver, kidney and brain. Chemically, lipids can be coupled to other molecules for the purpose of searching for direction to the target. The desired peptides, for example, hormones or neurotransmitters, and proteins such as antibodies or molecules are not peptides could be coupled to the liposomes in a chemical form. It is possible to remove the cells from the body and introduce the vector as a plasmid of naked DNA and then reimplant the transformed cells in the body. The naked DNA vector for gene therapy can be introduced into the desired host cells by methods known in the art. For example, tranfection, electroporation, microinjection, transduction, cell fusion, DEAE dextran, calcium phosphate precipitation, use of a gene gun or use of a transponder of DNA vectors [see for example Wi et al. , J. Bi ol. . Ch e. , 263: 963-961 (1992); Wu et al. , J. Bi or l. . Ch em. , 263: 14621-14624 (1988)] The Zcyto7 polypeptides can also be used to prepare antibodies that specifically bind Zcyto7 polypeptides. These antibodies can then be used to make anti-idiotypic antibodies. As used herein, the term "antibodies" includes polyclonal antibodies, antibodies, monoclonal antibodies, antigen fragments thereof such as F (ab ') 2 and Fab fragments and the like, including genetically engineered antibodies. Antibodies it is defined that they bind specifically if this binding to a Zcyto7 polypeptide with a Ka of more than equal to 107 / M. The affinity of a monoclonal antibody can be easily determined by one skilled in the art (see for example, Scatchard, ibid.) Methods for preparing polyclonal and monoclonal antibodies are well known in the art (see for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, (Second Edition) (Cold Spring Harbor, NY, 1989), and Hurrell, JGR, Ed., Monoclonar Hybridoma Antibodies: Techniques and Applications (CRC Press, Inc., Boca Raton, FL, 1982) As will be apparent to one skilled in the art, polyclonal antibodies can be generated from a variety of warm-blooded animals such as horses, cows, goats, sheep, dogs, chickens, rabbits, mice and rats. Zcyto polypeptide can be increased through the use of an adjuvant such as Freund's complete or incomplete adjuvant A variety of assays known to those skilled in the art can be used to detect anti-cancer. rpos that bind specifically to the Zcytol polypeptides. The essays of examples are described in detail in Antobodi is: A Labora t ory Manual, Harlow and Lane (Eds.), (Cold Spring Harbon Laboratory Press, 1988). Representative examples of these assays include: concurrent immunoelectrophoresis, radioimmunoassays, radioimmunoprecipitations, enzyme-linked inmubosorbent assays (ELISA), blot analysis, inhibition or competition assays, and intercalation assays. As will be apparent to one skilled in the art, polyclonal antibodies can be generated by inoculating a variety of warm-blooded animals such as horses, cows, goats, sheep, dogs, chickens, rabbits, mice, hamsters, cobalds and rats. with the Zcyto7 polypeptide or a fragment thereof. The immunogenicity of a Zcyto7 polypeptide can be increased by the use of an adjuvant, such as alum (aluminum hydroxide) or complete or incomplete Freund's adjuvant. Polypeptides useful for immunization also include fusion polypeptides such as Zcyto7 fusions or a portion thereof with an immunoglobin polypeptide or the amalatous binding protein. The immunogen of polypeptide can be a full-length molecule or a portion thereof. If the polypeptide portion is "hetene type", this portion can be advantageously linked or linked to a macromolecular carrier (such as limpet hemocyanin (KLH), bovine serum albumin (BSA) or tetanus toxoid for immunization. As used herein, the term "antibodies" includes polyclonal antibodies, polyclonal antibodies purified by affinity, monoclonal antibodies, and antigen-binding fragments, such as proteolytic fragments F. (ab ') 2. Genetically engineered intact antibodies or fragments, such as chimeric antibodies, Fv fragments, single chain antibodies and the like, as well as synthetic antigen binding peptides and polypeptides, are also included. Non-human antibodies can be humanized by grafting non-human CDRs into the human structure and constant regions, or by incorporating whole, non-human variable domains (optionally "covering" them with a human-like surface by replacing the exposed residues , where the result is a "capped" antibody). In some cases, the Humanized antibodies can retain non-human residues within the structure domains of the human variable region to improve the appropriate binding characteristics. Through the humanization of the antibodies, the biological half-life can be increased, and the potential for adverse involuntary reactions in administration to humans is reduced. Alternative techniques for generating or screening useful antibodies herein include in vitro exposure of limpocytes to the protein or Zcyto7 peptide, and selection of antibody display libraries in phage or similar vectors (e.g., through the use of the protein or peptide of certain Z cyt ol immobilized or marked). Genes coding for polypeptides having potential Zcyto7 polypeptide binding domains can be obtained by detecting the random peptide libraries displayed in the fab (phage display) or in bacteria such as E. col i. The nucleotide sequences encoding the polypeptides can be obtained in a number of ways such as through random mutagenesis and random synthesis of polynucleotides. These libraries of display of random peptides can be used to detect peptides that interact with a known target which can be a protein or polypeptide, such as a receptor ligand, or biological or synthetic macromolecule, or organic or inorganic substance. Techniques for creating and detecting these random peptide display libraries are known in the art (Ladner et al., U.S. Patent No. 5,223,409; Ladner et al., U.S. Patent No. 4,946,778: Ladner et al. , U.S. Patent No. 5,571,698); and random peptide display libraries and kits for detecting these libraries are commercially available, for example, from Clontech (Palo Alto, CA), Invitrogen Inc. (San Diego, CA), New England Biolabs, Ind. (Beverly , MA) and Pharmacia LKB Biotechnology Inc. (Piscataway, NJ). The random peptide display libraries can be detected using the Zcyto7 sequences. described herein to identify proteins that bind to Zcyto7. These "binding proteins" that interact with the Zcyto7 peptides can be used to label the cells; to isolate the homologous polypeptides by purification by affinity; drugs, toxins, radionuclides and the like can be conjugated directly or indirectly. These binding proteins can also be used in analytical methods such as detection of expression libraries and neutralization activity. Also, the binding proteins can be used for diagnostic assays, to determine the circulating levels of the polypeptides; to detect or quantify soluble polypeptides as a marker of the underlying pathology or disease. These binding proteins can also act as "antagonists" of Zcyto7 to block Zcyto7 binding and signal transduction in vitro and in vivo. The antibodies can also be generated by gene therapy. The animal is administered the DNA or RNA encoding Zcyto7 or an immunogenic fragment thereof so that the cells of the animals are transfected with nucleic acid and express the protein which in turn produces an immunogenic response. The antibodies that are then produced by the animal are isolated in the form of polyclonal or monoclonal antibodies.

Antibodies to Zcyto7 can be used to label cells expressing the protein, by affinity purification, within diagnostic assays to determine the circulation levels of soluble protein polypeptides, and as antagonists to block the binding junction (CLOCK WORD) and the translation of signals in vitro and in vivo. The hybrid-radiation correlation is a somatic cell genetic technique developed to construct contiguous high-resolution maps of mammalian chromosomes [Cox et al., Science 250: 245-250 (1990)]. The partial or complete knowledge of a gene sequence allows the design of PCR primers suitable for use with chromosomal radiation hybrid correlation panels. Commercially available radiation hybrid correlation panels covering the entire human genome, such as the Stanford G3 RH Panel and the GeneBridge 4 RH Panel panel (Research Genetics, Inc., Huntsville, AL), are available. These panels allow rapid localizations, based on PCR, chromosomal and gene sorting, sequence marked sites (STS), and other polymorphic markers and non-polymorphics within a region of interest. This includes the establishment of directly proportional physical distances between newly discovered genes of interest and previously correlated markers. Accurate knowledge of a gene position can be useful in a number of ways including: 1) determining whether a sequence is part of an existing contig and obtaining the surrounding, additional genetic sequences in various forms such as YAC-, BAC- or CDNA, 2) provide a possible candidate gene for a heriditable disease that allows the link to the same chromosomal region, and 3) for cross-referenced model organisms such as mice that may be helpful in determining what function a particular gene. The present invention also provides reagents that will find use in diagnostic applications. For example, the Zcyto7 gene has been correlated in chromosomal 5q31. A nucleic acid zone of Zcyto7 could be used to verify abnormalities on chromosome 5. For example, a probe comprising Zcyto7 DNA or RNA or a subsequence thereof can be used to determine if the Zcyto7 gene is present on chromosome 5q31 or if a mutation has occurred. Detectable chromosomal aberrations at the Zcyto7 gene locus include, but are not limited to, aneuploidy, gene copy number changes, insertions, deletions, restriction site changes, and rearrangements. These aberrations can be detected using the polynucleotides of the present invention by employing molecular genetic techniques, such as restriction fragment length polymorphism (RFLP) analysis, tandem repeat analysis (STR) employing PCR techniques, and other genetic linkage analysis techniques known in the art [Sambrook et al., ibid .; Ausubel, et. Al., Ibid .; Marian, A.J., Chest, 108: 255-265, (1995)]. Zcyto7 correlates in the 5q31 region which is a "gene cluster" that contains a group of cytokines and cytokine receptors. Clustered cytokines include IL-3, IL-4, IL-5, IL-13, GM-CSF, and M-CSF. This result authenticates zcyto7 as a cytokine. The invention is further illustrated by the following non-limiting examples: Example 1 Cloning of Zcyto7 Zcyto7 was identified from the defined expressed sequence tag (EST) by SEQ ID No. 3 for its homology to Interleukin-17. The cDNA clone was obtained from a library of Human cDNA from human fetal heart. The plasmid containing the cDNA was blotted on a 100 μg / ml ampicillin plate and 100 μg / ml LB methicillin. The cDNA insert was sequenced. The insert was determined to be 717 base pairs long with an open reading frame of 180 amino acids and a putative 20 amino acid signal peptide.

Example 2 Northern Transfer Analysis Transfers 1, 2, 3 of human multiple tissue (Clontech) were tested to determine the tissue distribution of Zcyto7. An EcoRI / NotI fragment containing the complete coding region of Zcyto7 was generated from clone EST582069 and used for the probe. A plasmid preparation of EST582069 was prepared from a overnight culture of 100 μg / ml ampicillin, LB, in 5 ml, 37 ° using the QIAprep Spin Miniprep Kit (Qiagen). 12 μl of the 100 μl were digested with 5 μl of H. buffer (Boehringer Mannheim), 12.5 units of EcoRI (Gibco BRL) and 12.5 units of Notl (New England Biolabs) in 50 μl of reaction at 37 ° C for 2 hours . Digestion was electrophoresed on a 0.7% TBE agarose gel and the fragment was cut. To obtain additional material, digestion was repeated under the same conditions as above, except that 24 μl of EST582069 was used in the second digestion. The second digestion was electrophoresed on a 0.7% TBE agarose gel and the fragment was cut. The DNA was extracted from the gel blocks with the gel extraction equipment (Qiagen). 135 ng of this DNA was labeled with P32 using the Multiprime DNA labeling scheme (Amersham) and the unincorporated radioactivity was removed with a NucTrap probe purification column (Stratagene). Northerns of multiple tissue were prehybridized and a marster transcription of human RNA 3 hours with 10 ml of ExpressHyb Solution (Contech) containing 1 mg of salmon sperm DNA that served 5 minutes and then cooled 1 minute and it was added to 10 ml of the ExpressHyb Solution, mixed and added to the transfers. Hybridization was carried out overnight at 65 ° C. The initial wash conditions were as follows: 2X SSC, 0.5% SDS, RT for 40 minutes with several solution changes then 0.1X SSC, 0.1% SDS at 50% for 40 minutes, 1 change of solution. The transfers were then exposed to the film at -80 ° C for 5 hours. There was cross-hybridization / background so that the blots were further washed at 55 ° C, then at 65 ° C with 0.1% X SSC, 0.1% SDS for 1 hour each. The spinal cord showed very high expression of Z cyt or l mRNA and the trachea showed weak mRNA expression. The size of the transcript was approximately 0.75 kb.

Example 3 Chromosomal Assignment and Zcyto7 Placement Zcyto7 was correlated to chromosome 5 using the "GeneBridge 1 Radiation Hybrid Panel" commercially available (Research Genetics, Inc., Huntsville, AL). The GeneBridge 4 Hybrid Radiation Panel contains the DNAs that are they can PCR each of the 93 clones of radiation hybrids, plus two control DNAs (the HFL donor and the A23 receptor). A publicly available WWW server (http: // www-genorne, wi.mit.edu/cgi-bin/cong ig / rhampper.pl) allows correlation to the Whitehead Institue / MIT Center for the search radiation hybrid map of the genome of the human genome (the radiation hybrid map "WICGR" of the human genome) that was constructed with the GeneBridge Hybrid Radiation Panel 4. For the correlation of Z cyt ol with the "RH Panel GeneBridge 4", they were put 20 μl reactions in a 96-well microtiter plate susceptible to PCR (Stratagene, La Jolla, CA) and used in a "RoboCycler Gradient 96" (Stratagene) thermal cycler. Each of the 95 PCR reactions consisted of 2 μl of KlenTaq 10X PCR ration buffer (CLONTECH Laboratories, INc, Palo Alto, CA), 1.6 μl of dNTPs mixture (2.5 mM each, PERKIN-ELMER, Foster City, CA), 1 μl of the homosentide primer of SEQ ID No. 4, 1 μl of the antisense primer of SEQ ID No. 5, 2 μl of "RediLoad" (Research Genetics, Inc., Huntsville, AL), 0.4 μl of the KlenTaq 50X Adventage polymerase mix (Clontech Laboratories, Inc.), 25 ng of the DNA of a single hybrid clone or control and x μl ddH20 for a total volume of 20 μl. The reaction was covered with an equal amount of mineral oil and sealed. The conditions of the PCR cycler were as follows. An initial cycle of 5 minutes of denaturation at 95 ° C, 35 cycles of 1 minute of desaturation at 95 ° C, 1 minute of fixation at 52 ° C and 1 minute of extension at 72 ° C, followed by a final extension cycle of 7 minutes at 72 ° C. The reactions were separated by electrophoresis on a 3% NuSieve GTG agarose gel (FMC Bioproducts, Rockland, ME). The results showed that Zcyto7 correlates to 490.89 cR from the top of linkage group 5 of the human chromosome in the WICGR radiation hybrid map. Regarding the centromere, its closest proximal marker was that of D5S413 and its closest distal marker was WI-5208. The use of surrounding markers placed Zcyto7 in the 5q31.3-q32 region on the integrated LDB chromosome 5 map (The Genetic Location Data Base, University of Southhampton, WWW server: http://cedar.genetics.soton.ac .uk / public html /).

Example 4 Construction of the Expression Vectors of Zcyto7 Two construction vectors of Zcyto7 were made in a FLAG amino acid sequence (SEQ ID No. 10) and inserted into the N-terminal or C-terminal ends of the Zcyto7 polypeptide. For the construction in which the FLAG amino acid sequence was linked to the N-terminus of Zcyto7, Zcyto7 PCR DNA fragment of 473 bp was generated with 1 μl of a 1/4 delution of the EST582069 plasmid preparation of Example 2 and 20 picomoles (pm) of the primer SEQ ID No. 6 and 20 pm of the primer SEQ ID No. 7. The PCR reaction was incubated at 94 ° C for 1 minute, and then ran for 5 cycles each individual cycle comprising 20 seconds at 94 ° C and 2 minutes at 64 ° C. This was followed for 22 cycles each cycle ranging from 20 seconds at 94 ° C and 2 minutes at 74 ° C. the reaction was terminated with incubation for 10 minutes at 74 ° C. 50 μl of the PCR reaction mixture was digested with 30 units of BamHl (Boehringer Mannheim) and 120 units of Xhol (Boehringer Mannheim) for 2 hours at 37 ° C. The digested reaction mixture is subjected to electrophoresis in a 1% TBE gel; the DNA band was excised with a blade and the DNA was extracted from the gel with the Qiaquick® gel extraction equipment (Qiagen). The excised DNA was subcloned into the nfpzp9 plasmid that has been cut with Bam and Zho. Nfpzp9 is a mammalian cell expression vector comprising an expression cartridge that contains the mouse metallothionein-1 promoter, a sequence encoding tissue plasminogen activator (TPOA), then the FLAG peptide (SEQ ID No. 10), then multiple restriction sites. These were followed by the human growth hormone terminator, an E. coli origin of replication and a mammalian selectable marker expression unit containing the SV40 promoter, enhancer and origin of replication; a dihydrofolate reductase gene (DHFR) and the SV40 terminator. For the construction of the Zcyto7 gene in which the FLAG of the C-term is inserted in the C-terminus of the Z cyt ol polypeptide, a PCR fragment of Zcyto7 of 543 bp was generated with 1 μl of a 1/4 dilution of the preparation of plasmid EST582069 described in Example 1 and 20 p.m. each of the primers SEQ ID No. 8 and SEQ ID No. 9. The PCR reaction was incubated at 94 ° C for 1 minute, then ran for 5 cycles, each cycle ranging from 20 seconds at 94 ° C and 2 minutes at 55 ° C. This was followed for 22 cycles each cycle comprised of 20 seconds at 94 ° C and 2 cycles at 74 ° C. The reaction was terminated with a final extension of 10 minutes at 74 ° C. The entire reaction mixture was run on a 1% TBE gel and the DNA was cut with a blade and the DNA was extracted using the QIAQUICK ™ gel extraction equipment. 20 μl of the 35 μl recovered were digested with 10 units of BamHl (Boehringer Mannheim) and 10 units of EcoRl (Gibco BRL) for 2 hours at 37 ° C. The digested PCR mixture was subjected to electrophoresis in a 1% TBE gel. The DNA band was cut with a blade, and the DNA was extracted from the gel using the QIAquick ™ gel extraction kit (Qiagen). The extracted DNA was subcloned into the cfpzp9 plasmid that has been cut with EcoRl and BamHl. The plasmid cfpzp9 is a mammalian expression vector that contains an expression cartridge having the mouse ionein-1 metallot promoter, multiple restriction sites for the insertion of the coding sequences, a sequence encoding the FLAG peptide, SEQ ID No. 10, a terminator codon, a human growth hormone terminator, an E. coli origin of replication, a mammalian selectable marker expression unit having an SV 40 promoter, a intensifier and origin of replication, and a DHFR gene and the SV40 terminator. Using the antibodies to the FLAG polypeptides, the FLAG-tagged Zcyto7 polypeptides can be separated from a supernatant fluid of cells.

Example 5 Cloning of murine Zcyto7 The mouse Zcyto7 was identified from EST, SEQ ID No. 14, by its homology to human Zcyto7. The cDNA clone was discovered in a murine embryo cDNA library in which the embryos were between 13.5 and 14.5 days old. The cDNA was delivered as an agar sting containing E. coli transfected with the plasmid having the cDNA of interest and then plotted on a 100 μg / ml ampicillin plate, 25 μg / ml LB methicillin. The cDNA insert in EST660242 was sequenced. It was determined that the insert is from 785 base pairs with an open reading frame of 180 amino acids and a putative signal peptide of 20 amino acids. The sequences are defined by SEQ ID No. 11 and SEQ ID No. 12.

Example 6 Zcyto7 tissue distribution in cattle The Northern blot of multiple tissue, from mouse (Clontech, Palo Alto, CA), transfer in mouse Northern blot (Clontech), a Northern blot of mouse embryos, and a spot blot of the mouse spinal cord were tested to test the tissue distribution of murine Zcyto7. RNA from mouse embryos was isolated from the embryos of mice that were six days from the date of fertilization using the isolation kit (POLI (A) PURÉ® mRNA (ambion). 100 mg of each mouse embryo was smoothed in 1 ml of lysis buffer, homogenized, processed in a batch method according to the manufacturer's protocol.For Northern blotting, 2 ug of RNA was loaded on 1.5% agarose, gel formaldehyde 2.2.M. The gel was run at 60 V for 4 hours and 30 minutes. The RNA was transferred overnight on a Nytran membrane that was pre-wetted in 20 X SSC. The RNAs were crosslinked on the membrane by UV light and baked at 80 ° C for one hour. The RNA of the spinal cord of the mouse was then prepared with the isolation equipment (POLI (A) PURÉ® mRNA (ambion).) The transfer in spot of the spinal cord of 1 mouse was done by covering a spot with 1, 2 and 3 ul of RNA at a concentration of 1 μg RNA / ul on a Nytran membrane A fragment of No tl / £ coRI containing the complete Zcyto7 coding region was generated from the clone containing SEQ ID No: 12 (referred to later as the clone of SEQ ID No: 12) and was used for the probe. The plasmid preparation of the clone of SEQ ID No: 12 was prepared from 5 ml overnight ampicillin culture of 100 ug / ml of LB at 37 ° C using the QIAPREP SPIN MINIPREP kit (Quiagen) were digested4. 66 ug with 8 ul of high shock absorber (Boehringer Mannheim), 20 units of No t l (Biolabs) and 20 units of £ coRI (Gibco BRL) in 80 ul reaction at 37 ° C for 2 hours. Digestion is subjected to electrophoresis in a 1.0% TBE gel and the cut fragment. The DNA was extracted from the gel block with a QIAQUICK® gel extraction kit (Qiagen). The 98.8 mg of this fragment was labeled with P32 using the MULTIMPRIME® DNA marking system (Amersham) and the unincorporated radioactivity was removed with a NUCTRAP® probe purification column (Estratagene). The two Northern preparations and the two spot transfer preparations were prehybridized for 3 hours at 65 ° C as follows. 1 mg of salmon was hybridized for 5 minutes, cooled 1 minute, mixed with 10 ml of EXPRESSHYB® solution and added to the spots. Hybridization was carried out overnight at 65 ° C. The initial washing conditions were as follows: 2 X SSC, 01% SDS for 40 minutes at room temperature, then 0.1 X SSC, 0.1% SDSal for 40 minutes at 50 ° C. Transfers to the film were transposed overnight at minus 80 ° C. Northern blots and mouse blot were further washed with 0.1% X SSC, 0.1% SDS at 60 ° C to remove the background. The spot transfer of the mouse spinal cord was again washed at a high severity with 0. 1 X SSC, 0.1% SDS at 65 ° C to confirm the previous results.

Results: The expression of Z cyt or mouse l was seen in the spinal cord, the submaxillary gland and the epidimos. The mouse embryos showed Zcyto7 expression starting on day 12, reaching the maximum on day 16 and ending on day 17 from the date of fertilization. The size of the transcript was 1 kb.

Example 7 Proliferation of chondrocytes using Zcyto7 A chondrocyte proliferation assay was performed to determine the effect that Zcyto7 would have on the proliferation of chondrocytes. The test was done with 20% confluent cultures. As a control vehicle, bovine serum albumin was added to a culture of chondrocytes instead of Zcyto7. The assay measured the incorporation of 3H-thymidine from nascent DNA into chondrocytes, Wahl et al. , Mol. Cel l. Bi ol. . 5: 5016-5025 (1988).

Results: A 3.5-9 fold stimulation of primary chondrocyte proliferation was seen in the exposure of chondrocyte cultures at 1 μg / ml of Zcyto7. The stimulation of the chondrocytes by Zcyto7 was seen with multiple preparations of the • protein and was seen through lines of species. In contrast to this, the control experiment using BSA did not result in chondrocyte stimulation.

Example 8 Production of glycosaminoglycan by chondrocytes treated with Zcyto7 A 20% confluent culture of chondrocytes was prepared and Zcyto7 was applied at a concentration of 1 ug / ml. In a second experiment in addition to Zcyto7 IL-lβ was applied to the culture of the cells. In a control group, BSA was added to the culture of chondrocytes. The level of glycosaminoglycan production (GAG) by the culture of chondrocytes was then determined using a dye binding assay of 1, 9-dimethymethylene blue, dye binding assay, Fardale et al. , Bi och em. Bi ophys. Acta 555: 173-177 (1987).

Results: The chondrocytes that were cultured with Zcyto7 showed a 50% increase in the presence in stable state of GAG in the culture of chondrocytes. Furthermore, when the chondrocytes were co-cultured with both Zcyto7 and interlucin-lβ (IL-1), the production of GAG by the chondrocytes was increased 2.5 compared with the culture of chondrocytes with either Zcyto7 or IL-lß. While the cultured cells to which BSA was added did not show an increased production of GAG.

Example 9 Stimulation of osteoblasts by Zcyto7 The CCC4 cell line is a line of osteoblast-like cells derived from faint p53 mice. The CCC4 line was transfected with a plasmid containing an inducible ceric response element (SER) that drives the expression of luciferase. The simulation of the SER and in this way the expression of luciferase indicates that the chemical entity is probably going to stimulate the osteoblasts.

CCC4 cells were cultured in the presence of 1 μg of Zcyto7 ml culture medium. As a control, each of BSA, fibroblast growth factor (FGF) and platelet derived growth factor (PDGF) were added to different cultures of CCC4 cells. The BSA is a negative control and the FGF and the PDGF were positive controls since they are known to promote the proliferation of osteoblasts. Luciferase activity was detected by the addition of 40 ul of promising luciferase substrate using an integrated 2-second reading in Labsistes LUMINOSKAN® " Results: Zcyto7 as well as FGF and PGDF stimulate luciferase expression in this assay indicating that it stimulates osteoblasts. BSA vehicle control was negative in this trial.

Example 10 Effect of Zcyto7 on the growth of fibroblasts Congruent cultures of fetal lung, human lung, thermal fibroblast cells were inoculated with Zcyto7 to determine the effects of Z cyt ol in the growth of fibrolasts. FGF was used as a positive control and BSA as a negative control.

Results: Zcyto7 had no effect on the growth of fibroblasts.

Example 11 Effect of Zcyto7 on the growth of BaF3 cells BaF3 cells, a line of murine pre-B cells dependent on IL-3 to proliferate, were washed several times with a base medium and plated in a 96-well plate, each well containing approximately 5500 cells / well. The cells were treated with either 1 ug / ml of Z cyto l or l-2pg / ml of IL-3 or with a combination of both Zcyto7 and IL-3. Also in a separate experiment, 0.1-10 mg / ml of TGBβ was added to the wells instead of Zcyto7. After incubation of the assay plate at 37 ° C and 5% C02 for 3-6 days, 20 ul of ALAMAR blue was added to each well and the plate incubated at 37 ° C for 15-24 hours. The plate is then read with a fluorometer with the excitation wavelength of 544 m and the emission wavelength of 590 m. The assay was also labeled by the eye for the stimulation or inhibition of cell proliferation before the addition of ALAMAR blue. The base medium contained RPMI 1640 + 10% HIA-FBS + L-glutamine + Na pyruvate.

Results: Zcyto7 and TGFβ significantly inhibited the proliferation driven by IL-3 of BaF3 cells. However, when the antibodies were neutralized to TGFβ, Zcyto7 inhibition of the proliferation of BaF3 cells was eliminated together with Z cyt or l.

Example 12 Effect of Zcyto7 on the growth of TF-1 cells A human leukemia cell line that is dependent on GM-CSF or IL-lβ were washed several times with the base medium and then placed in a 96-well plate with each well containing approximately 7,000 cells / well. The cells were cultured with 1 μg / ml of Zcyto7 and 100-200 μg / ml of IL-lβ. Also in a separate experiment, TGFβ was added to the wells instead of Zcyto7.

After incubation of the assay plate at 37 ° C and 5% C02 for 3-5 days, 20 ul of ALAMAR blue was added to each well and the plate was incubated at 73 ° C for 15-24 hours. The plate then with a fluorometer with excitation wavelength of 544 nm and emission wavelength of 590 nm. The assay was also assessed by the eye for stimulation and inhibition of cell proliferation before the addition of ALAMAR blue. The base medium contained RPMI 1640 + 10% HIA-FBS + 1-glutamine + Na pyruvate.

Results: The stimulation of IL-lβ of TF-1 cells was inhibited by both Zcyto7 and TGF-β. The concentration of Zcyto7 at which the inhibition of proliferation occurred was greater than 200ng / ml; and the concentration of TGF-β at which the inhibition of proliferation occurred was approximately 50 pg / ml.

LIST OF SEQUENCES (1) GENERAL INFORMATION (i) APPLICANT: ZymoGenetics, Inc. 1201 Eastlake Avenue East Seattle Avenue East WA USA 98102 (ii) TITLE OF THE INVENTION: FACTOR 7 CYTOKINE TYPE OF MAMMALS (iii) SEQUENCE NUMBER: 10 (iv) ADDRESS OF CORRESPONDENCE (TO) RECIPIENT: ZymoGenetics, Inc. (B) STREET: 1201 Eastlake Avenue East (C) CITY: Seattle (D) STATE: WA (E) COUNTRY: UAE (F) ZIP CODE (ZIP) : 98102 (v) LEGIBLE FORM IN COMPUTER: (A) TYPE OF MEDIA: Flexible disk (B) COMPUTER: compatible with IBM PC (C) OPERATING SYSTEM: DOS (D) PROGRAM: FastSEQ for Windows Version vi) CURRENT APPLICATION DATA (A) APPLICATION NUMBER: (B) SUBMISSION DATE: (C) CLASSIFICATION vii) DATE OF PREVIOUS APPLICATION (A) NUMBER OF APPLICATION: (B) DATE OF SUBMISSION: (viii) ATTORNEY / AGENT INFORMATION: (A) NAME: Lunn, Paul G (B) REGISTRATION NUMBER: 32,743 (C) REFERENCE NUMBER / DOCUMENT: 97-15PC (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 206-442-6627 (B) TELEFAX: 206-442-6678 (C) TELEX: (2) INFORMATION FOR SEQ ID 1.0: 1: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 736 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: cDNA (ix) CHARACTERISTIC: (A) NAME / KEY: Sequence encoding (B) LOCATION: 57 ... 596 (D) OTHER INFORMATION: (xi) DESCRIPTION FOR SEQ. ID No: l: GAATTCGGCA CGAGGAGGCG GGCAGCAGCT GCAGGCTGAC CTTGCAGCTT GGCGGA ATG 59 Met 1 GAC TGG CCT CAC AAC CTG CTG TTT CTT CTT ACC Ap TCC ATC TTC CTG 1U7 Asp Trp Pro His Asn Leu Leu Phe Leu Leu Thr He Ser He Phe Leu 5 10 15 GGG CTG GGC CAG CCC AGG AGC CCC AAA AGC AAG AGG AAG GGG CAA GGG 155 Gly Leu Gly Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly 20 25 30 CGG CCT GGG CCC CTG GCC CCT GGC CCT CAC CAG GTG CCA CTG GAC CTG 203 Arg Pro Gly Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu 35 40 45 GTG TCA CGG ATG AAA CCG TAT GCC CGC ATG GAG GAG TAT GAG AGG AAC 251 Val Ser Arg Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn 50 55 50 65 ATC GAG GAG ATG GTG GCC CAG CTG AGG AAC AGC TCA GAG CTG GCC CAG 299 He Glu Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing Gln 70 '75 80 AGA AAG TGT GAG GTC AAC pG CAG CTG TGG ATG TCC AAC AAG AGG AGC 347 Arg Lys Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser 85 90 95 CTG TCT CCC TGG GGC TAC AGC ATC AAC CAC GAC CCC AGC CGT ATC CCC 395 Leu Ser Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro 100 105 110 GTG GAC CTG CCG GAG GCA CGG TGC CTG TGT CTG GGC TGT GTG AAC CCC 443 Val Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro 115 120 125 pC ACC ATG CAG GAG GAC CGC AGC ATG GTG AGC GTG CCG GTG pC AGC 491 Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser 130 135 140 145 CAG Gp CCT GTG CGC CGC CGC CTC TGC CCG CCA CCG CCC CGC ACA GGG 539 Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly 150 155 160 CCT TGC CGC CAG CGC GCA GTC ATG GAG ACC ATC GCT GTG GGC TGC ACC 587 Pro Cys Arg Gln Arg Ala Val Met Glu Thr He Wing Val Gly Cys Thr 165 170 175 TGC ATC pC TGAATCACCT GGCCCAGAAG CCAGGCCAGC AGCCCGAGAC CATCCTCCT 645 C? S He Phe .180 TGCACCpTG TGCCAAGAAA GGCCTATGAA AAGTAAACAC TGACTpTGA AAGCCAGAAA 705 AAAAAAAAAA AAAAAAAAp CCTGCGGCCG C 736 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 180 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (v) TYPE OF FRAGMENT: internal (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 2 Met Asp Trp Pro His Asn Leu Leu Phe Leu Leu Thr lie Ser lie Phe 1 5 10 15 Leu Gly Leu Gly Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln 20 25 30 Gly Arg Pro Gly Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp 35 40 45 Leu Val Ser Arg Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg 50 55 60 Asn lie Glu Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing 65 70. 75 80 Gln Arg Lys Cys Gu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg 85 90 95 Ser Leu Ser Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He 100 105 110 Pro Val Asp Leu Pro Glu Ala Arg Cys Leu Cys Leu Gly Cys Val Asn 115 120 125 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 130 135 140 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr 145 150 155 160 Gly Pro Cys Arg Gln Arg Wing Val Met Glu Thr He Wing Val Gly Cys 165 170 175 Thr Cys He Phe 180 (2) INFORMATION FOR SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 397 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 3: AGGCGGGCAN AGCTGCAGGC TGACCpGCA GCpGGCGGA ATGGACTGGC CTCACMCCT 60 GCTGTpcp CpACCApT CCATCpCCT GGGGCTGGGC AGCCAGGAGC CCCAAAAGCA 120 AGAGGAAGGG GCAAGGGCGG CCTGGGCCCN TGGCCTGGCC TCACCAGGTG CCACTGGACC 180 TGGTGTCACG GATGAAACCG TATGCCCGCA TGGAGGAGTA TGAGAGGAAC ATCGAGGAGA 240 TGGTGGCCCA GCTGAGGAAC AGCTCANAAG CTGGCCCAGA GAAAGTGTGA GGTCAACpG 300 CAGCTGTGGA TGTCCAACAA GAAGGAGCCT GTCTCCCpG GGGCTACAAG CATCAACCAC 360 CGACCCCAGC CGTATCCCCG TGGGACC? TG CCGGGAC 397 (2) INFORMATION FOR SEQ ID NO:: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13265 (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 4 TTACCATTTC CATCTTCC (2) INFORMATION FOR SEQ ID NO: 5 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13266 (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 5: CCCTTCCTCT TGCTTTTG (2) INFORMATION FOR SEQ ID NO: 6 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13326 xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 6: CAAGGATCCC AGCCCAAGGAG CCCCAAAAG (2) INFORMATION FOR SEQ ID NO: 7 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13330 (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 7 GACCTCGAGT CAGAAGATGC AGGTGCAGCC (2) INFORMATION FOR SEQ ID NO: 8 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13325 xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 8: GTCGAATTCA TGGACTGGCC TCACAACCTG (2) INFORMATION FOR SEQ ID NO: 9 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (vii) IMMEDIATE SOURCE: (B) CLON: ZC13327 (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 9: GAAGGATCCG AAGATGCAGG TGCAGCC (2) INFORMATION FOR SEQ ID NO: 10: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 10 amino acids (B) TYPE: amino acids (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID: yr Lys Asp Asp Asp Asp Lys Gly Ser 5 10 INFORMATION FOR SEQ ID NO: 11 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 692 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: Other (ix) CHARACTERISTIC: (A) NAME / KEY: Codified Sequence (B) LOCATION: 50 ... 589 (D) OTHER INFORMATION: (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 11: GGGGpCCTG GCGGGTGGCA GCTGCGGGCC TGCCGCCTGA CTTGGTGGG ATG GAC TGG 58 Met Asp Trp 1 CCG CAC AGC CTG CTC pC CTC CTG GCC ATC TCC ATC pC CTG GCG CCA 106 Pro His Ser Leu Leu Phe Leu Leu Ala lie Ser He Phe Leu Ala Pro 5 10 '15 AGC CAC CCC CGG AAC ACC AAA GGC AAA AGA AAA GGG CAA GGG AGG CCC 154 Ser His Pro Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln Gly Arg Pro 20 25 30 35 AGT CCC pG GCC CCT GGG CCT CAT CAG GTG CCG CTG GAC CTG GTG TCT 202 Ser Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser 40 45 50 CGA GTA AAG CCC TAC GCT CGA ATG GAA GAG TAT GAG CGG AAC CTT GGG 250 Arg Val Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn Leu Gly 55 60 65 GAG ATG GTG GCC CAG CTG AGG AAC AGC TCC GAG CCA GCC AAG AAG AAA 298 Glu Met Val Ala Gln Leu Arg Asn Ser Ser Glu Pro Ala Lys Lys Lys 70 75 80 TGT GAA GTC AAT CTA CAG CTG TGG pG TCC AAC AAG AGG AGC CTG TCC 346 Cys Glu Val Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg Ser Leu Ser 85 90 95 CCA TGG GGC TAC AGC ATC AAC CAC GAC CCC AGC CGC ATC CCT GCG GAC 394 Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Wing Asp 100 105 110 115 pG CCC GAG GCG CGG TGC CTA TGT pG GGT TGC GTG AAT CCC pC ACC 442 Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr 120 125 130 ATG CAG GAG "GAC CGT AGC ATG GTG AGC GTG CCA GTG pC AGC CAG GTG 490 Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val 135 140 145 CCG GTG CGC CGC CGC CTC TGT CCT CAA CCT CCT CGC CCT GGG CCC TGC 538 Pro Val Arg Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro Gly Pro Cys 150 155 160 CGC CAG CGT GTC GTC ATG GAG ACC ATC GCT GTG GGT TGC ACC TGC ATC 586 Arg Gln Arg Val Val Met Glu Thr He Wing Val Gly Cys Thr Cys He 165 170 175 pC TGAGCCAACC ACCAACCCGG TGGCCTCTGC AACAACCCTC CCTCCCTGCA CCCACT 645 Phe 180 GTGACCCTCA AGGCTGATAA ACAGTAAACG CTGpcpíG TAAAGGA 692 (2) INFORMATION FOR SEQ ID NO: 12 i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 180 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (v) TYPE OF FRAGMENT: internal (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 12: Met Asp Trp Pro His Ser Leu Leu Phe Leu Leu Ala He Ser He Phe 1 5 10 15 Leu Ala Pro Ser His Pro Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln 20 25 30 Gly Arg Pro Ser Pro Leu Ala Pro Gly Pro His Gln Val Pro Leu Asp 35 40 45 Leu Val Ser Arg Val Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg 50 55 60 Asn Leu Gly Glu Met Val Wing Gln Leu Arg Asn Ser Ser Glu Pro Wing 65 70 75 80 Lys Lys Lys Cys Glu Val Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg 85 90 95 Ser Leu Ser Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He .100 105 110 Pro Wing Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn 115 120 125 Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe 130 135 140 Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro 145 150 155 160 Gly Pro Cys Arg Gln Arg Val Val Met Glu Thr He Wing Val Gly Cys 165 170 175 Thr Cys He Phe 180 (2) INFORMATION FOR SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 497 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: cDNA (xí) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 13: GGGGpCCTG GCGGGTGGCA GCTGCGGGCC TGCCGCCTGA CpGGTGGGA TGGACTGGCC 60 GCACAGCCTG CTCTTCCTCC TGGCCATCTC CATCpCCTG GCGCCAAGCC ACCCCCGGAA 120 CACCAAAGGC AAAAGAAAAG GGCAAGGGAG GCCCAGTCCC pGGCCCCTG GGCTCATCAG 180 GTGCCGCTGG ACCTGGTGTC TCGAGTAAAG CCCTACGCTC GAATGGAAGA GTATGAGCGG 240 AACCTTGGGG AGATGGTGGC CCAGCTGAGG AACAGCTCCG AGCCAGCCAA GAAGAAATGT 300 GAAGTCAATC TACAGCTGTG GpGTCCAAC AAGAGGAGCC TGTCCCCATG GGGCTACAGC 360 ATCAACCACG ACCCCAGCCG CATCCCTGCG GACpGCCCG AGGCGCGGTG CCTATGTTTG 420 GGpGCGTGA ATCCCTTCAC CATGCAGGAG GACCGTAGCA TGGTGAGCGT GCCAGTGpC 480 AGCCAGGTGC CGGTGCG 497 (2) INFORMATION FOR SEQ ID NO: 14: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 14 Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gn Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn lie Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met VaT Ser Val Pro Val Phe Ser Gln Val Pro 115 120 125 (2) INFORMATION FOR SEQ ID NO: 15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID: Gln Pro Arg Wing Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Gu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Glu Leu Wing Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 16: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 16: Gn Pro Arg Pro Pro Lys Wing Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Pro Wing Pro Gly Pro Val Val Leu Asp Leu Val Ser Arg 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu -. 35 -. 40 45 Met Val Ala Gln Leu Arg Asn Ser Ser Glu Leu Ala Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asp His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met . 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Val Pro Phe Ser Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 17: (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 17: Gln Pro Arg Pro Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Wing 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Ala Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100. 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro 115 120 125 Val Arg Arg - Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 2) INFORMATION FOR SEQ ID NO: 11 (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear ii) TYPE OF MOLECULE: protein xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 18: Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1. ' 5 10 15 Pro Leu Pro Wing Pro Pro His Gln Val Pro Leu Asp Leu Val Wing Arg 20 25 30 Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Being Glu Leu Ala Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115 .120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 2) INFORMATION FOR SEQ ID NO: 19 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 19: Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Glu Leu Wing Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met .100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Val Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 20: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 20: Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Glp Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Glu Leu Wing Gln Arg Lys Cys fifty - . 50 - 55 60 Glu Val Asn Leu Gn Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Gl? Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Leu Val Met Glu Thr He Wing Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 21: i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 21: Gln Pro Arg Pro Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Glp Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Ala Gln Leu Arg Asn ' Being Ser Glu Leu Ala Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Phe Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 INFORMATION FOR SEQ ID NO: 22 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 22: Gln Pro Arg Pro Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1. 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Gly Arg. "20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Ala Gln Leu Arg Asn Ser Ser Glu Leu Ala Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gn Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr'lle Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 23: Gln Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Ser 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Glu Leu Wing Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115. "120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys lie Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 24: 61n Pro Arg Ser Pro Lys Val Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Gln Leu Arg Asn Ser Glu Leu Wing Gln Arg Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75. 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 61n Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 25: Gln Pro Arg Val Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Met Lys Pro Tyr Wing Arg Met Gl u Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 Met Val Wing Glp Leu Arg Asn Being Ser Glu Leu Wing Glp Arg Lys Cys 50 55. 60 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115 120 125 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 130 135. 140 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 26: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 97 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 26: Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser 1 5 10 15 Pro Trp Gly Tyr Ser He A = n His Asp Pro Ser Arg He Pro Val Asp 20 25 30 Leu Pro Glu Ala Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr 35. "40 45 Met Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val 50 55 60 Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys 65 70 75 80 Arg Gln Arg Wing Val Met Glu Thr He Wing Val Gly Cys Thr Cys He 85 90 95 Phe (2) INFORMATION FOR SEQ ID NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 100 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi DESCRIPTION FOR THE SEQUENCE: SEQ ID No: 27 Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gl n Gly Arg Pro Gly Pro 1 5 10 15 Leu Wing Pro Gly Pro Hi s Gln Val Pro Leu Asp Leu Val Ser Arg Met 20 25 30 Lys Pro Tyr Al a Arg Met Gl u Glu Tyr Glu Arg Asn He Gl u Glu Met 35 •. 40 45 Val Al a Gln Leu Arg Asn Be Ser Glu Leu Ala Gln Arg Lys Cys Gl u 50 55 60 Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro Trp 65 70 75 80 Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu Pro 85 90 95 Gl u Ala Arg Cys 100 INFORMATION FOR SEQ ID NO: 28 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 28: Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly Pro 1 5 10 15 Leu (2) INFORMATION FOR SEQ ID NO: 29: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 29: Arg Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He G u 1 5 10 15 Glu (2) INFORMATION FOR SEQ ID NO: 30: (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 16 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 30: sn Hi s Asp Pro Ser Arg He Pro Val Asp Leu Pro Glu Ala Arg Cys 1 5 10 15 (2) INFORMATION FOR SEQ ID NO: 31: (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 31: Pro Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys 1 5 10 15 Arg Gln Arg (2) INFORMATION FOR SEQ ID NO: 32: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 47 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 32: Pro Arg Ser Pro Lys Ser Lys Arg Lys Gly Gl n Gly Arg Pro Gly Pro 1 5 10 15 Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg Met 20 25 30 Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 35 40 45 (2) INFORMATION FOR SEQ ID NO: 33: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 70 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 33: Arg Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu 1 .- 5 10 15 Glu Met Val Wing Gln Leu Arg Asn Being Ser Glu Leu Wing Gln Arg Lys 20 25 30 Cys Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser 35 40 45 Pro Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp 50 55 60 Leu Pro Glu Ala Arg Cys 65 70 (2) INFORMATION FOR SEQ ID NO: 34: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 61 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 34: Asn His Asp Pro Be Arg He Pro Val Asp Leu Pro Glu Ala Arg Cys 1 5 10 15 Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met Gln Glu Asp Arg Ser 20 25 30 Met Val Ser Val Pro Val Phe Ser Gln Val Pro Val Arg Arg Arg Leu 35 40 45 Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg 50 55 60 (2) INFORMATION FOR SEQ ID NO: 35: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 73 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 35: Asn His-Asp Pro Ser Arg He Pro Val Asp Leu Pro Glu Wing Arg Cys 1 5 10 15 Leu Cys Leu Gly Cys Val Asp Pro Phe Thr Met Gln Glu Asp Arg Ser 20 25 30 Met Val Ser Val Pro Val Phe Ser Gln Val Pro Val Arg Arg Arg Leu 35 40 45 Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg Ala Val Met 50 55 60 Glu Thr He Wing Val Gly Cys Thr Cys 65 70 (2) INFORMATION FOR SEQ ID NO: 36: (i) CHARACTERISTICS OF THE SEQUENCE (A) LENGTH: 158 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 36: Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly Pro Leu 1 5 10 15 Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg Met Lys 20. 25 30 Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu Met Val 35 40 45 Wing Gln Leu.Arg Asn Ser Ser Glu Leu Wing Gln Arg Lys Cys Glu Val 50 55 60 Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro Trp Gly 65 70 75 80 Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu Pro Glu 85 90 95 Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met Gln Glu 100 105 110 Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro Val Arg 115 120 125 Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg 130 135 140 Wing Val Met Glu Thr He Wing Val Gly Cys Thr Cys He Phe 145 150 155 (2) INFORMATION FOR SEQ ID NO: 37: i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 154 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 37: Be Lys Arg Lys Gly Gln Gly Arg Pro Gly Pro Leu Wing Pro Gly Pro 1 5 10 15 His Gln Val Pro Leu Asp Leu Val Ser Arg Met Lys Pro Tyr Wing Arg 20 25 30 Met Glu Glu Tyr Glu Arg Asn He Glu Glu Met Val Ala Gln Leu Arg 35 40 45 Asn Be Ser Glu Leu Ala Gln'Arg Lys Cys Glu Val Asn Leu Gln Leu 50 55 60 Trp Met Ser Asn Lys Arg Ser Leu Ser Pro Trp Gly Tyr Ser He Asn 65 70 75 80 His Asp Pro Ser Arg He Pro Val Asp Leu Pro Glu Wing Arg Cys Leu 85 90 95 Cys Leu Gly Cys Val Asn Pro Phe Thr Met Gln Glu Asp Arg Ser Met 100 105 Val Val Ser Val Phe Ser Gln Val Pro Val Arg Arg Arg Leu Cys 115 120 125 Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg Ala Val Met 61u 130 135 140 Thr He Ala -Val Gly Cys Thr Cys He Phe 145 150 2) INFORMATION FOR SEQ ID NO: 38 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 151 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 38: Lys Gly Gln Gly Arg Pro Gly Pro Leu Wing Pro Gly Pro His Gln Val 1, 5 10 15 Pro Leu Asp Leu Val Ser Arg Met Lys Pro Tyr Ala Arg Met Glu Glu. -20 25 30 Tyr Glu Arg Asn He Glu Glu Met Val Wing Gln Leu Arg Asp Being Ser 35 40 45 Glu Leu Wing Glp Arg Lys Cys Glu Val Asn Leu Gln Leu Trp Met Ser 50 55 60 Asn Lys Arg Ser Leu Ser Pro Trp Gly Tyr Ser He Asn His Asp Pro 65 70 75 80 Being Arg He Pro Val Asp Leu Pro Glu Wing Arg Cys Leu Cys Leu Gly 85 90 95 Cys Val Asn Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val Ser Val 100 105 110 Pro Val Phe Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro Pro Pro 115 120 125 Pro Arg Thr Gly Pro Cys Arg Gln Arg Ala Val Met 61u Thr He Ala 130 135 140 Val Gly Cys Thr Cys He Phe 145 150 (2) INFORMATION FOR SEQ ID NO: 39: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 160 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 39: His Pro Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln Gly Arg Pro Ser 1 5 10 15 Pro Leu Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg 20 25 30 Val Lys Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn Leu Gly Glu 35 40 45 Met Val Ala Gln Leu Arg Asn Ser Ser Glu Pro Ala Lys Lys Lys Cys 50 55 60 Glu Val Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg Ser Leu Ser Pro 65 70 75 80 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Wing Asp Leu 85 90 95 Pro Glu Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 100 105 110 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Be Gln Val Pro 115.- 120 125 Val Arg Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro Gly Pro Cys Arg 130 135 140 Gln Arg Val Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 160 (2) INFORMATION FOR SEQ ID NO: 40: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 158 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 40: Arg Asn Thr Lys Gly Lys Arg Lys Gly Gln Gly Arg Pro Ser Pro Leu 1 5 10 15 Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg Val Lys 20 25 30 Pro Tyr Ala Arg Met Glu Glu Tyr Glu Arg Asn Leu Gly Glu Met Val 35 40 45 Wing Gln Leu Arg Asn Ser Ser Glu Pro Wing Lys Lys Lys Cys Glu Val 50 55 60 Asn Leu Gln Leu Trp Leu Ser Asn Lys Arg Ser Leu Ser Pro Trp Gly 65 70 75. 80 Tyr Ser He Asn His Asp Pro Ser Arg He Pro Wing Asp Leu Pro Glu 85 90 95 Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met Gn Glu 100 105 110 Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro Val Arg 115 120 125 Arg Arg Leu Cys Pro Gln Pro Pro Arg Pro Gly Pro Cys Arg Gln Arg 130 135 140 Val Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 145 150 155 (2) INFORMATION FOR SEQ ID NO: 41: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 153 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 41: Lys Arg Lys Gly Gln Gly Arg Pro Gly Pro Leu Wing Pro Gly Pro His 1 5 10 15 Gln Val Pro Leu Asp Leu Val Ser Arg Met Lys Pro Tyr Wing Arg Met 20 25 30 Glu Glu Tyr Glu Arg Asn He Glu Glu Met Val Wing Gln Leu Arg Asn 35 40 45 Be Ser Glu Leu Wing Gln Arg Lys Cys Glu Val Asn Leu Gln Leu Trp 50 55 60 Met Ser Asn Lys Arg Ser Leu Ser Pro Trp Gly Tyr Ser He Asn His 65 70 75 80 Asp Pro Ser Arg He Pro Val Asp Leu Pro Glu Wing Arg Cys Leu Cys 85 90 95 Leu Gly Cys Val Asn Pro Phe Thr Met Gln Glu Asp Arg Ser Met Val 100 105 110 Ser Val Pro Val Phe Ser Gln Val Pro Val Arg Arg Arg Leu Cys Pro 115 120 125 Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg Wing Val Met Glu Thr 130 135 140 He Wing Val Gly Cys Thr Cys He Phe 145 150 (2) INFORMATION FOR SEQ ID NO: 42: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 128 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID No: 42: Met Lys Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu 1 5 10 15 Met Val Wing Gln Leu Arg Asn Ser Ser Glu Leu Wing Gln Arg Lys Cys 20 25 30 Glu Val Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro 35. - 40 45 Trp Gly Tyr Ser He Asn His Asp Pro Ser Arg He Pro Val Asp Leu 50 55 60 Pro Glu Ala Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met 65 70 75 80 Gln Glu Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro 85 90 95 Val Arg Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg 100 105 110 Gln Arg Ala Val Met Glu Thr He Ala Val Gly Cys Thr Cys He Phe 115 120 125 (2) INFORMATION FOR SEQ ID NO: 43: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 157 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: simple (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION FOR THE SEQUENCE: SEQ. ID or: 43: Arg Ser Pro Lys Ser Lys Arg Lys Gly Gln Gly Arg Pro Gly Pro Leu 1 5 10 15 Wing Pro Gly Pro His Gln Val Pro Leu Asp Leu Val Ser Arg Met Lys • 2.0 25 30 Pro Tyr Wing Arg Met Glu Glu Tyr Glu Arg Asn He Glu Glu Met Val 40 45 Ala Gln Leu Arg Asn Ser Ser Glu Leu Ala Gln Arg Lys Cys Glu Val 50 55 60 Asn Leu Gln Leu Trp Met Ser Asn Lys Arg Ser Leu Ser Pro Trp Gly 65 70 75 80 Tyr Ser He Asn His Asp Pro Ser? Rg He Pro Val Asp Leu Pro Glu 85 90 95 Wing Arg Cys Leu Cys Leu Gly Cys Val Asn Pro Phe Thr Met Gln Glu 100 105 110 Asp Arg Ser Met Val Ser Val Pro Val Phe Ser Gln Val Pro Val Arg 115 120 125 Arg Arg Leu Cys Pro Pro Pro Pro Arg Thr Gly Pro Cys Arg Gln Arg 130 135 140 Wing Val Met Glu Thr He Wing Val Gly Cys Thr Cys He 145 150 155

Claims (24)

1. An isolated polynucleotide encoding a mammalian Zcyto7 polypeptide or variants thereof.

2. The isolated polynucleotide according to claim 1, characterized in that the polynucleotide encodes a polypeptide selected from the group SEQ ID No: 2, SEQ ID No: 12, SEQ ID Nos: 14-27 and SEQ ID Nos: 36-43 .

3. A polynucleotide according to claim 2, characterized in that the polypeptide encoded by the polynucleotide is a polypeptide defined by SEQ ID NOS: 15-25 wherein the amino termini of the polypeptides start at either the amino acid residue 3, an arginine , amino acid residue 7, a cerin, amino acid residue 8, a lysine; amino acid residue 10, a lysine residue of amino acid 33, a methionine.

4. A polynucleotide according to claim 2, characterized in that the polypeptide encoded by the polynucleotide is a polypeptide defined by SEQ ID No: 2, 12, 14-25 and 36 to 42, wherein the amino acid sequences terminate in the isoleucines at amino acid residue 179 of SEQ ID No: 2 , at amino acid residue 159 of SEQ ID NO: 14-25, which corresponds to the amino acid residue of 157 of SEQ ID NO: 36, amino acid residue 153 of SEQ ID NO: 37, amino acid residue 150 of SEQ ID NO: 38, amino acid residue 159 of SEQ ID NO: 39, amino acid residue 157 of SEQ ID No. 40, amino acid residue 152 of SEQ ID N: 42, amino acid residue 127 of SEQ ID NO : 42

5. An isolated polynucleotide encoding a peptide or polypeptide having at least 15 amino acid residues comprised of a portion having an epitope of a polypeptide of SEQ ID Nos: 2, 14-27 and 36-43.

6. The isolated polynucleotide according to claim 5 characterized by the peptide or polypeptide is selected from the group consisting of SEQ ID NOS: 28-35.

7. The polynucleotide according to claim 5 or 6, characterized in that the peptide or polypeptide is fused to a carrier polypeptide or another carrier molecule.

8. A polynucleotide of which encodes a peptide or polypeptide that is at least 90%, 95% or 99% identical to the polypeptides encoded by the nucleotides of claims 1-7.

9. An expression vector characterized in that it comprises the following operably linked elements: a transcription promoter; a DNA segment defined by claim 1-8; and a transcriber terminator.

10. An expression vector comprising the following operably linked elements: (a) a transcription promoter (b) a DNA segment encoding a chimeric polypeptide, wherein the chimeric polypeptide consists essentially of a first portion and a second portion linked by a link peptide, the first portion that is comprised by a mammalian polypeptide, the polypeptide which is the amino acid sequences of SEQ ID nOS: 2, 12, 14-43 and the second portion which is a second polypeptide or protein. (c) a transcription terminator.

11. An isolated mammalian Z cyt ol polypeptide or variants thereof.

12. E7L Zcyto7 polypeptide isolated according to claim 11, characterized in that it is selected from the amino acid sequence group consisting of SEQ ID Nos: 2, 12, 14-27 and 36-43.

13. A polypeptide according to claim 12, characterized in that the polypeptide is a polypeptide defined by SEQ ID No: 14-25, wherein the amino termini of the polypeptides are modified and start at either amino acid residue 3, an argininin , amino acid residue 7, a cerin; amino acid residue 8, a lysine; amino acid residue 10, a lysine or amino acid residue 33, a methionine.

14. A polypeptide according to claim 12, characterized in that the polypeptide is a polypeptide defined by SEQ ID Nos: 2, 12, 14-25 and 36-42 wherein the amino acid sequences terminate in the isoleucines at amino acid residue 179 of SEQ ID NO: 2, at amino acid residue 159 of SEQ ID Nos: 14-25, which corresponds to amino acid residue 157 of SEQ ID NO: 36, amino acid residue 153 of SEQ ID No: 37, amino acid residue 159 of SEQ. ID no; 38, amino acid residue 159 of SEQ ID NO: 39, amino acid residue 157 of SEQ ID NO: 48, amino acid residue 152 of SEQ ID NO: 42, amino acid residue 127 of SEQ ID NO: 42.

15. An isolated peptide or polypeptide characterized in that it has at least 14 amino acid residues comprised of a portion having epitope of a polypeptide of a Zcyto7 polypeptide.

16. The peptide or polypeptide according to claim 15 characterized in that the peptide or polypeptide is a portion having epitope of a polypeptide of SEQ ID Nos: 2, 14-27 and 36-43.

17. The isolated peptide or polypeptide according to claim 16 characterized in that the peptide or polypeptide is selected from an amino acid sequence group consisting of SEQ ID NOS: 28-35.

18. An isolated peptide or polypeptide, characterized in that it is 90% & 95 or 99% identical to the peptides or polypeptides of claims 12-17.

19. The isolated peptide or polypeptide according to claims 12-17 characterized in that it has an amino acid sequence by the addition, deletion and / or replacement of one or more amino acid residues and that it maintains the biological activity of the peptide or polypeptide.

20. An antibody characterized in that it binds specifically to a mammalian polypeptide, the polypeptide that is defined by the amino acid sequence of claims 12-20.

21. A method for producing an antibody that binds to a peptide or polypeptide of claims 12-19, characterized in that it comprises inoculating an animal with the peptide or polypeptide or with a nucleic acid encoding the peptide or polypeptide, wherein the animal produces antibodies to the peptide or polypeptide; and isolate the antibody.

22. The antibody according to claim 21, characterized in that the antibody is polyclonal or monoclonal.

23. Antibody fragments, single chain antibodies or humanized antibodies of the antibodies of claims 20 and 21.

24. An anti-idiotypic antibody of an antibody of claims 21-23.