MXPA01009178A - Mammalian cytokines;related reagents and methods - Google Patents

Abstract

Purified genes encoding cytokine from a mammal, reagents related thereto including purified proteins, specific antibodies, and nucleic acids encoding this molecule are provided. Methods of using said reagents and diagnostic kits are also provided.

Description

MAMMALS CYCOTINES: RELATED REAGENTS AND METHODS FIELD OF THE INVENTION The present invention relates to compositions related to proteins that function in the control of the biology and physiology of mammalian cells, for example, cells of a mammalian immune system. In particular, it provides purified genes, proteins, antibodies, related reagents and useful methods, for example, to regulate the activation, development, differentiation and function of various cell types, including hematopoietic cells.

BACKGROUND OF THE INVENTION Recombinant DNA technology generally refers to the technique of integrating genetic information from a donor source to vectors for further processing, such as introduction into a host, whereby the transferred genetic information is copied and / or expressed in the new environment. Commonly, genetic information exists in the form of complementary DNA (cDNA) derived from messenger RNA (mRNA) which codes for a desired protein product. The carrier is often a plasmid that has the ability to incorporate cDNA for subsequent replication in a host, and in some cases, to actually control the expression of the cDNA and thus direct the synthesis of the product encoded in the host. For some time, it has been known that the immune response of a mammal is based on a series of complex cellular interactions, called "immune networks". See, for example, Paul (1998) Fundamental Immunoloqy (4th ed.) Raven Press, NY. The recent research has provided new insights into the internal workings of this network. While it is clear that much of the response, in fact, revolves around the network-like interactions of lymphocytes, macrophages, granulocytes and other cells, immunologists now generally hold the view that soluble proteins, known as lymphokines, cytokines or monocins, play a decisive role in the control of these cellular interactions. Consequently, there is considerable interest in the isolation, characterization and mechanisms of action of cellular modulating factors, the understanding of which will lead to important advances in the diagnosis and therapy of numerous medical abnormalities, for example, disorders of the immune system. Some of these factors are hematopoietic growth factors, for example, granulocyte colony stimulating factor (G-CSF). See, for example, Thomson (ed. 1998) The Cvtokine Handbook (3rd ed.) Academic Press, San Diego; Mire-Sluis and Thorpe (ed. 1998) Cvtokines Academic Press, San Diego, Metcalf and Nicola (1995) The Hematopoietic Colonv Stimulatinq Factors. Cambridge University Press; and Aggarwal and Gutterman (1991) Human Cvtokines Blackwell Pub.

Lymphokines apparently act as intermediates in cellular activities in a variety of ways. They have been shown to support the proliferation, growth and / or differentiation of pluripotent hematopoietic stem cells in large numbers of progenitors comprising various cell lines, forming a complex immune system. Appropriate and balanced interactions between cellular components are necessary for a healthy immune response. Different cell lines often respond differently when lymphokines are administered together with other agents. The cell lines especially important for the immune response include two classes of lymphocytes: B cells, which can produce and secrete immunoglobulins (proteins with the ability to recognize and bind foreign matter to effect their removal), and T cells of several subclasses, which secrete lymphokines and induce or suppress B cells and several other cells (including other T cells), which make up the immune network. These lymphocytes interact with many other cell types. Research for the better understanding and treatment of various immune disorders has been hampered by the general inability to keep cells of the immune system in vitro. Immunologists have discovered that the culture of these cells can be effected through the use of T cell supernatants and other cells, which contain various growth factors, including many of the lymphokines.

From the foregoing, it is evident that the discovery and development of new lymphokines, for example, related to G-CSF and / or IL-6, could contribute to new therapies for a wide range of degenerative or abnormal conditions that directly or indirectly involve the immune system and / or hematopoietic cells. In particular, the discovery and development of lymphokines that enhance or enhance the beneficial activities of known lymphokines would be extremely convenient. The present invention provides novel compositions of interleukins and related compounds, and methods for their use.

BRIEF DESCRIPTION OF THE INVENTION The present invention is directed to mammalian interleukin-B60 (IL-B60), eg, primate or rodent, and to its biological activities. It includes nucleic acids that code for polypeptides themselves, and methods for their production and use. The nucleic acids of the invention are characterized, in part, by their homology to complementary DNA sequences (cDNAs) described therein, and / or by functional assays for activities of the cytokine or growth factor type, e.g., G-CSF (see Nagata (1994) in Thomson, The Cvtokine Handbook, 2nd Ed., Academic Press, San Diego), and / or IL-6 (see Hirano (1994) in Thomson, The Cvtokine Handbook, 2nd Ed., Academic Press, San Diego). In addition, polypeptides, antibodies and methods for the use thereof are provided, including the use of nucleic acid expression methods. Methods for modulating or intervening in the control of a physiology dependent on growth factor, or an immune response are provided. The present invention is based, in part, on the discovery of a new cytokine sequence that exhibits significant sequence and structural similarity to G-CSF and IL-6. In particular, it provides genes for primate, for example, human, and rodent, for example, mouse, which encode a protein whose mature size is approximately 198 amino acids. Functional equivalents exhibiting significant sequence homology will be available from other mammalian species, e.g., cows, horse and rat. In addition, the present invention identifies a second associated component of a complex. Compositions related to the combination of components in the complex are provided, together with methods of use. In one embodiment, the invention provides a substantially pure or recombinant polypeptide, comprising the mature protein portion of SEQ ID NO: 2 or 4. Preferably, the polypeptide is: detectably labeled; non-glycosylated; denatured; attached to a solid substrate; conjugated to another chemical portion, or in a sterile composition. The forms of kits include those comprising the polypeptide and: a compartment comprising the polypeptide; or with instructions for the use or disposal of reagents in the equipment.

Binding compounds include those which comprise an antigen binding site of an antibody, which specifically binds to the described polypeptide. The linking compound may also be in a kit comprising: a compartment comprising the linking compound; or with instructions for the use or disposal of reagents in the equipment. The invention further provides a method for the production of an antigen: antibody complex, which comprises contacting, under appropriate conditions, a primate IL-B60 polypeptide with an antibody that binds specifically or selectively to the polypeptide of the invention, thereby enabling the complex is formed. Nucleic acid modalities include an isolated or recombinant polynucleotide encoding the mature protein portion of SEQ ID NO: 2 or 4. In other embodiments, the invention provides an isolated soluble complex comprising the mature protein portion of SEQ ID NO. : 2 or 4, and the mature protein portion of SEQ ID NO: 12 or 13. Preferably the complex: comprises a recombinant polypeptide of SEQ ID NO: 2, 4, 12 or 13; it is detectably labeled; it is in a regulated solution; It is in a sterile solution. Equipments are provided that contain said complex and: a compartment comprising the complex; or instructions for the use or disposal of equipment reagents. Binding compounds are provided which comprise an antigen binding site of an antibody that specifically binds to the soluble complex, but not to the mature polypeptide of SEQ ID NO: 12 or 13. Equipment is provided that comprises the linking compound and: a compartment comprising the linking compound; or instructions for the use or disposal of equipment reagents. Methods are provided, for example, for the production of an antigen: antibody complex, comprising contacting, under appropriate conditions, a primate complex comprising the polypeptides IL-B60 and CLF-1, with an antibody that selectively or specifically binds to an isolated soluble complex comprising the mature protein portion of SEQ ID NO: 2 or 4, and the mature protein portion of SEQ ID NO: 12 or 13, thus allowing the complex to be formed. The nucleic acid modalities include a recombinant or isolated nucleic acid encoding the mature protein portion of SEQ ID NO: 2 or 4, and the mature protein portion of SEQ ID NO: 12 or 13. The invention further provides a composition of material selected from: an isolated polypeptide comprising at least seven amino acids identical to segments of SEQ ID NO: 2 or 4; a substantially pure or recombinant polypeptide comprising at least two distinctive non-overlapping segments of at least five amino acids identical to the segments of SEQ ID NO: 2 or 4; a natural sequence polypeptide comprising mature SEQ ID NO: 2 or 4; or a fusion polypeptide comprising the sequence IL-B60. In certain embodiments, distinctive identity segments that do not overlap include: one of at least eight amino acids; one of at least five amino acids and one second of at least six amino acids; at least three segments of at least four, five and six amino acids, or one of at least twelve amino acids. In other embodiments, the polypeptide of the composition of matter: is the polypeptide which: comprises a mature sequence of Table 1; it is a non-glycosylated form of IL-B60; it's from a primate, just like a human; comprises at least seventeen amino acids of SEQ ID NO: 2 or 4; exhibits at least four non-overlapping segments of at least seven amino acids of SEQ ID NO: 2 or 4; it is a natural allelic variant of IL-B60; it has a length of at least about 30 amino acids; exhibits at least two non-overlapping epitopes, which are specific for a primate IL-B60; it is glycosylated; has a molecular weight of at least 30 kD with natural glycosylation; is a synthetic polypeptide, is attached to a solid substrate; it is conjugated to another chemical portion; it is a substitution of 5 times or less of natural sequence; it is a variant of deletion or insertion of a natural sequence; or which further comprises: at least seven amino acids identical to segments of SEQ ID NO: 12 or 13; at least two non-overlapping distinctive segments, of at least five amino acids identical to segments of SEQ ID NO: 12 or 13; a natural sequence polypeptide comprising SEQ ID NO: 12 or 13 mature; or a primate CLF'1. In further preferred embodiments, the composition comprises: a substantially pure IL-B60 and CLF-1; a sterile IL-B60 polypeptide comprising the mature protein of SEQ ID NO: 2 or 4; or the described polypeptide and a carrier, wherein the carrier is: an aqueous compound, including water, saline and / or pH regulator; and / or formulated for oral, rectal, nasal, topical or parenteral administration. The invention provides fusion polypeptides comprising: mature protein sequence of Table 1; a detection or purification queue, including a FLAG, His6, or Ig sequence; or sequence of another cytokine receptor family protein, including CLF-1. Team modalities include those comprising the polypeptide of the composition and: a compartment comprising the protein or polypeptide; or instructions for the use or disposal of equipment reagents. The invention further provides methods for the use of the described polypeptides: for labeling the polypeptide, which comprises labeling the polypeptide with a radioactive label; for separating the polypeptide from another polypeptide in a mixture, comprising running the mixture in a chromatography matrix, thereby separating the polypeptides; to identify a compound that selectively binds to the polypeptide, which comprises incubating the compound with the polypeptide under appropriate conditions, thereby causing the component to bind to the polypeptide; or to conjugate the polypeptide to a matrix, which comprises deriving the polypeptide with a reacting reagent, and conjugating the polypeptide to the matrix. Related linking compounds include those which comprise an antigen binding site of an antibody that specifically or selectively binds to a natural polypeptide, as described above, wherein: the linking compound is in a container; the IL-B60 polypeptide is from a human; the linking compound is an Fv, Fab, or Fab2 fragment; the linking compound is conjugated to another chemical portion; or the antibody: is produced against a mature polypeptide of Table 1; is produced against a mature IL-B60; is produced for a purified human IL-B60; is immunoselected; it is a polyclonal antibody; binds to a denatured IL-B60; exhibits a Kd to antigen of at least 30 μM; it is attached to a solid substrate, including a bead or plastic membrane; it is in a sterile composition; or is detectably labeled, including a radioactive or fluorescent label. Equipments comprising said binding compound and: compartment comprising the linking compound are provided; or instructions for the use or disposal of equipment reagents. Methods for the production of an antigen: antibody complex are provided, which comprise contacting, under appropriate conditions, a primate IL-B60 polypeptide with a described antibody, thus allowing the complex to be formed. Preferably, in the method: the complex is purified from other cytokines; the complex is purified of another antibody; the contact is with a sample comprising a cytokine; the contact allows the quantitative detection of the antigen; the contact is with a sample comprising the antibody; or the contact allows the quantitative detection of the antibody.

In another embodiment the invention includes a composition comprising: a sterile binding compound, as described, or the linking compound and a carrier, wherein the carrier: wherein the carrier is: an aqueous compound, including water, saline and / or pH regulator; and / or formulated for oral, rectal, nasal, topical or parenteral administration. Nucleic acid modalities include an isolated or recombinant nucleic acid encoding the described polypeptide, wherein: IL-B60 is from a human; or the nucleic acid: encodes a peptide sequence antigen of Table 1; encodes a plurality of peptide antigen sequences from Table 1; encodes a plurality of peptide antigen sequences from Table 4; exhibits identity over at least thirteen nucleotides to a natural cDNA encoding the segment; it is an expression vector; further comprises an origin of replication; it is from a natural source; comprises a detectable label; comprises synthetic nucleotide sequence; is less than 6 kb, preferably less than 3 kb; it's from a primate; comprises a natural full-length coding sequence; is a hybridization probe for a gene that codes for IL-B60; or is a PCR primer, PCR product, or mutagenesis primer. Preferred embodiments include those where the isolated or recombinant nucleic acid is in a cell or tissue. The cell can be: a prokaryotic cell; a eukaryotic cell; a bacterial cell; a yeast cell; an insect cell; a mammalian cell; a mouse cell; a primate cell; or a human cell.

Equipments comprising the described nucleic acid and: a compartment comprising the nucleic acid are provided; a compartment further comprising a primate IL-B60 polypeptide; or. instructions for the use or disposal of equipment reagents. The invention further provides methods for forming a duplex with a polynucleotide described above, comprising contacting the polynucleotide with a probe that hybridizes, under severe conditions, to at least 25 contiguous nucleotides of the coding portion of SEQ ID NO: 1, 3, or which encodes SEQ ID NO: 12 or 13 natural; thus forming the pair. In another aspect, the invention provides a nucleic acid which: hybridizes under washing conditions of 30 minutes at 30 ° C and less than 2 M salt, to the coding portion of SEQ ID NO: 1; or exhibits identity on a stretch of at least about 30 nucleotides to a primate IL-B60. In preferred embodiments, wash conditions that are at 45 ° C and / or at 500 mM salt; or at 55 ° C and / or at 150 mM salt; or the stretch is at least 55 nucleotides, for example, at least 75 nucleotides. Methods are provided, for example, of modulating the physiology or development of a cell or tissue culture cells, comprising contacting the cell with an agonist or antagonist of a mammalian IL-B60; or contacting the cell with an agonist or antagonist of a complex comprising mammalian IL-B60 and CLF-1. Additionally, the invention provides a method for increasing the secretion of: an IL-B60 sequence, which comprises expressing the polypeptide with CLF-1; or a CLF-1, which comprises expressing CLF-1 with an IL-B60 sequence. In preferred embodiments of the method, the increase is at least 3 times, 5X, 7X, I0X, or more; or the expression is of a recombinant nucleic acid encoding one or both of the polypeptide and CLF-1. The invention further provides a screening method for a receptor that binds an isolated soluble complex comprising the mature protein portion of SEQ ID NO: 2 or 4, and the mature protein portion of SEQ ID NO: 12 or 13, which comprises contacting the complex with a cell expressing the receptor, under conditions that allow the complex to bind to the receptor, thus forming a detectable interaction. Preferably, the interaction produces a physiological response in the cell. Other embodiments of the invention include, for example, an isolated soluble complex comprising at least 6 amino acids of the mature protein portion of SEQ ID NO: 2 or 4, and: at least 6 amino acids of the mature protein portion of SEQ ID NO: 12 or 13; or at least 6 amino acids of the mature protein portion of CNTF-R. Said complex may, for example, comprise a recombinant polypeptide of SEQ ID NO: 2 or 4 mature; comprising a recombinant polypeptide of SEQ ID NO: 12 or 13 mature; comprise a recombinant mature CNTF-R polypeptide; comprising both a recombinant polypeptide of SEQ ID NO: 2 or 4 mature, and a recombinant polypeptide of SEQ ID NO: 12 or 13 mature; comprising both a mature SEQ ID NO: 2 or 4 recombinant polypeptide and a mature recombinant CNTF-R polypeptide; be detectably labeled; be in a regulated solution; or be in a sterile solution. Preferred embodiments include those that: comprise a mature IL-B60 polypeptide; comprise a mature CLF-1 polypeptide; comprise a mature CNTF-R polypeptide; exhibit at least four non-overlapping segments of at least seven amino acids of SEQ ID NO: 2 or 4; exhibit epitopes of both primate L-B60 and primate CLF-1; exhibit epitopes of both primate L-B60 and primate CNTF-R; they are not glycosylated; they are attached to a solid substrate; they are conjugated to another chemical portion; or comprise a detection or purification tail, including a FLAG, His6, or Ig sequence. Equipment is provided, for example, comprising the complex and: a compartment comprising the complex, and / or instructions for the use or disposal of reagents in the equipment. Fusion polypeptides are provided, including, for example, an isolated or recombinant polypeptide comprising: a first segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4, and a second segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 12 or 13 mature; at least two non-overlapping distinctive segments, of at least five amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and a third segment comprising at least seven amino acids identical to the segments of SEQ ID NO. : 12 or 13 mature; at least one segment comprises at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and two distinctive non-overlapping segments of at least five amino acids identical to the segments of SEQ ID NO: 12 or 13 mature; a first segment comprises at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4, and a second segment comprises at least seven amino acids identical to the mature primate CNTF-R segments; at least two distinctive non-overlapping segments of at least five amino acids identical to the segments of mature SEQ ID NO: 2 or 4, and a third segment comprising at least seven amino acids identical to the CNTF-R segments of mature primate; or at least one segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and two distinctive non-overlapping segments of at least five amino acids identical to the CNTF-R segments of mature primate. Certain modalities include those in which the distinctive identity segments do not overlap: they include one of at least eight amino acids; they include one of at least five amino acids and one second of at least six amino acids; they include at least three segments of at least four, five and six amino acids, or include one of at least twelve amino acids. Other embodiments include those that: comprise a mature IL-B60 sequence; comprise a mature CLF-1 sequence; comprise a mature CNTF-R sequence; exhibit at least four non-overlapping segments of at least seven amino acids of SEQ ID NO: 2 or 4; they have a length of at least about 30 amino acids; exhibit epitopes of both primate IL-B60 and primate CLF-1; exhibit epitopes of both primate IL-B60 and primate CNTF-R; they are not glycosylated; have a molecular weight of at least 30 kD; they are a synthetic polypeptide; they are attached to a solid substrate; they are conjugated to another chemical portion; or comprise a detection or purification tail, including a FLAG, His6, or Ig sequence. Other embodiments include a composition comprising: substantially pure combination of IL-B60 and CLF-1; substantially pure combination of IL-B60 and CNTF-R; a sterile polypeptide described above; or the polypeptide described above and a carrier, wherein the carrier is: an aqueous compound, including water, salt and / or pH buffer; and / or formulated for oral, rectal, nasal, topical or parenteral administration. A kit comprising a polypeptide as described is provided, and: a compartment comprising the polypeptide and / or instructions for the use or disposal of reagents in the kit. In addition, methods are provided, for example, for making an antibody that recognizes a complex as described, which comprise inducing an immune response in an animal with the complex; for immunoselecting antibodies, comprising contacting a population of antibodies with a complex as described, and separating antibodies that bind from those that do not bind; or to formulate a composition, comprising mixing a complex as described with a carrier. Binding compounds are provided, for example, which comprise an antigen binding site of an antibody, which antibody binds specifically to a described complex, but not to any of the mature polypeptides of SEQ ID NO: 2, 4, 12, 13 , or CNTF-R. Certain modalities include those in which: the binding compound is: in a container; an Fv, Fab, or Fab2 fragment; or conjugated to another chemical portion; or the antibody: is produced against a substantially pure complex of IL-B60 with CLF-1; is produced against a substantially pure complex of IL-B60 with CNTF-R; is immunoselected; it is a polyclonal antibody; exhibits a Kd to antigen of at least 30 μM; it is attached to a solid substrate, including a bead or plastic membrane; it is in a sterile composition; or is detectably labeled, including a radioactive or fluorescent label. Additional embodiments include a composition comprising: a sterile binding compound as described, or the linking compound as described and a carrier, wherein the carrier is: an aqueous compound, including water, salt and / or pH buffer; and / or formulated for oral, rectal, nasal, topical or parenteral administration. With the binding composition there is provided a kit comprising the linking compound and: a compartment comprising the linking compound; or instructions for the use or disposal of equipment reagents. In addition there are provided methods for the production of an antigen: antibody complex, comprising contacting under appropriate conditions, a primate complex comprising: polypeptides IL-B60 and CLF-1; or polypeptides IL-B60 and CNTF-R; with an antibody as described, thus allowing the complex to be formed. Preferably, in the method, the complex is purified from other cytokines; the complex is purified from another antibody; the contact is with a sample comprising a cytokine; the contact allows the quantitative detection of the antigen; the contact is with a sample comprising the antibody; or the contact allows the quantitative detection of the antibody. Various nucleic acids are provided, for example, an isolated or recombinant nucleic acid: encoding the amino acid portions described above; encoding the amino acid portions as described, and comprising a segment of at least 30 contiguous nucleotides of SEQ ID NO: 1 or 3; which will co-express a segment of at least seven contiguous amino acids of SEQ ID NO: 2 or 4, and a segment of at least seven contiguous amino acids of SEQ ID NO: 12 or 13; or which will co-express a segment of at least seven contiguous amino acids of SEQ ID NO: 2 or 4, and a segment of at least seven contiguous amino acids of CNTF-R. Preferred nucleic acids include those that, for example: encode IL-B60 from a human; encode CLF-1 of a human; encode CNTF-R from a human; they are a vector of expression; they also comprise an origin of replication; they comprise a detectable label; comprise a synthetic nucleotide sequence; or are less than 6 kb, preferably less than 3 kb. A cell comprising the recombinant nucleic acid is provided, for example, wherein the cell is: a prokaryotic cell; a eukaryotic cell; a bacterial cell; a yeast cell; an insect cell; a mammalian cell; a mouse cell; a primate cell; or a human cell. Various nucleic acid kits are provided, for example, comprising the nucleic acid and: a compartment comprising the nucleic acid; a compartment further comprising a primate IL-B60 polypeptide; a compartment further comprising a primate CLF-1 polypeptide; a compartment further comprising a primate CNTF-R polypeptide; or instructions for the use or disposal of equipment reagents. Methods are also provided, for example, for making an acid, double nucleic acid, comprising contacting said nucleic acid with a complementary nucleic acid, under appropriate conditions, thereby forming said duplex; to express a polypeptide, which comprises expressing the nucleic acid, thereby producing the polypeptide; or to transfect a cell, comprising contacting said cell under appropriate conditions with the nucleic acid, thus transfecting the cell. In an alternative embodiment, the invention provides an isolated or recombinant nucleic acid encoding at least 5 contiguous amino acids of SEQ ID NO: 12, 13, or primate CNTF-R: hybridized under 30 minute 30 ° wash conditions C and less than 2 M salt, to the coding portion of SEQ ID NO: 1; or exhibits identity on a stretch of at least about 30 nucleotides, to a primate IL-B60. Preferred embodiments include: the isolated nucleic acid, wherein: the contiguous amino acids number at least 8; the washing conditions are at 45 ° C and / or at 500 mM salt; or the stretch is at least 55 nucleotides; or the recombinant nucleic acid, wherein: contiguous amino acids number at least 12; the washing conditions are at 55 ° C and / or at 150 mM salt; or the stretch is at least 75 nucleotides. The invention in particular provides methods for the modulation of the physiology or development of a cell or tissue culture cells, comprising contacting the cell with an agonist or antagonist of a complex comprising mammalian IL-B60 and: CLF- 1; or CNTF-R. It also provides methods for the production of the proteins, for example, the production of a described complex, which comprise co-expressing a recombinant IL-B60 with a recombinant CNTF-R or CLF-1; increase the secretion of an IL-B60 polypeptide, which comprises expressing the polypeptide with CLF-1 or CNTF-R; or increase the secretion of a CLF-1 polypeptide, which comprises expressing CLF-1 with an IL-B60. Typically, the increase is at least 3 times; or the expression is of a recombinant nucleic acid encoding one or both of the polypeptide and CLF-1. Further provided are methods for screening for a receptor that binds to the described complex, comprising contacting the complex with a cell expressing the receptor, under conditions that allow the complex to bind to the receptor, thereby forming a detectable interaction. Preferably, the interaction produces a physiological response in the cell.

DETAILED DESCRIPTION OF THE PREFERRED MODALITIES All references cited herein are incorporated herein by reference, to the same extent as if each publication or individual patent application were specifically and individually indicated to be incorporated by reference. 1. General The present invention provides amino acid sequences and DNA sequences that encode several mammalian proteins that are cytokines, for example, which are secreted molecules that can act as intermediates of a signal between immune or other cells. See, for example, Paul (1998) Fundamental Immunoloqy (4th ed.) Raven Press, N. Y. Full-length cytokines, and fragments, or antagonists, will be useful, for example, in the physiological modulation of cells expressing a receptor. It is possible that IL-B60 has either stimulatory or inhibitory effects on hematopoietic cells, including, for example, lymphoid cells, such as T cells, B cells, natural killer cells (NK), macrophages, dendritic cells, hematopoietic progenitors, etc. The proteins will also be useful as antigens, for example, immunogens, for the production of antibodies for various epitopes in the protein, both linear and conformational epitopes. A sequence encoding IL-B60, of a human genomic sequence, was identified. The molecule was designated hulL-B60. Also described is a rodent sequence, eg, mouse. The human gene encodes a protein of the small soluble cytokine type, of approximately 198 amino acids. The predicted signal sequence psort is probably about 17 residues, and would run from the Met to about Ala. See Table 1 and SEQ ID NO: 1 and 2. IL-B60 exhibits structural motifs characteristic of a member of the long chain cytokines. Compare, for example, IL-B60, G-CSF, and IL-6, sequences available from GenBank. The closest confrontation is with CT-1, oncostatin M, and CNTF. See also Table 2.

TABLE 1 Nucleic acid (SEQ ID NO-1) encoding IL-B60 of a primate, eg, human. Predicted signal decomposition site is indicated. Nucleotide 375 can be A. The translated amino acid sequence is SEQ ID NO: 2 ccgagcgaaa aaaacctgcg agtgggcctg gcggatggga ttattaaagc ttcgccggag 60 ccgcggctcg ccctcccact ccgccagcct ccgggagagg agccgcaccc ggccggcccg 120 gccccagccc catggacctc atg tta cgagcagggg actcgtgggg g gcg tgc ctg 176 Met Leu Ala Leu Cys -15 tgc aeg gtg ctc tgg falls ctc ect gea gtg cea gct ctc aat cgc here 224 Cys Thr Val Leu Trp His Leu Pro Ala Val Pro Ala Leu Asn Arg Thr -10 -5 -1 ggg gac cea ggg ect ggc ccc tec ate cag aaa acc tat gac ctc acc 272 Gly Asp Pro Gly Pro Gly Pro Ser lie Gln Lys Thr Tyr Asp Leu Thr 10 15 20 cgc tac ctg gag falls ca ctc cgc age ttg gct ggg acc tae ctg aac 320 Arg Tyr Leu Glu His Gln Leu Arg Ser Leu Wing Gly Thr Tyr Leu Asn 25 30 35 tac ctg ggc ccc ect t aac gag cea gac t aac ect ccc cgc ctg 363 Tyr Leu Gly Pro Pro Phe Asn Glu Pro Asp Phe Asn Pro Pro Arg Leu 40 45 50 ggg gea gag act ctg ccc agg gcc act gtt gac ttg gag gtg tgg cga 416 Gly Ala Glu Thr Leu Pro Arg Ala Thr Val Asp Leu Glu Val Trp Arg 55 60 65 age ctc aat gac aaa ctg cgg ctg acc ac tac gac gcc tac age 454 Ser Leu Asn Asp Lys Leu Arg Leu Thr Gln Asn Tyr Glu Ala Tyr Ser 70 75 80 falls ctt ctg tgt tac ttg cgt ggc ctc aac cgt cag gct gcc act gee 512 His Leu Leu Cys Tyr Leu Arg Gly Leu Asn Arg Gln Wing Wing Thr Wing 85 90 95 100 gag ctc cgc cgc age ccg gcc falls tte tgc acc age ctc cag ggc ctg 560 Glu Leu Arg Arg Ser Leu Wing His Phe Cys Thr Ser Leu Gln Gly Leu 105 110 H5 ctg ggc age acc gcg ggc gcc aeg gea gcc ccg ggc tac cea ctg ccc 608 Leu Gly Ser He Wing Gly Val Met Wing Wing Leu Gly Tyr Pro Leu Pro 120 125 130 cag ccg ceg ect ggg act gaa ccc act cgg acc ccc gcc ect gcc cae 656 Gln Pro Leu Pro Gly Thr Glu Pro Thr Trp Thr Pro Gly Pro Ala His 135 140 145 agt gac tte ctc cag aag atg gac gac tte tgg ctg ctg aag gag ctg 704 Be Asp Phe Leu Gln Lys Met Asp Asp Phe Trp Leu Leu Lys Giu Leu 150 155 160 cag acc tgg ctg tgg cgc teg gcc aag gac tte aac cgg cec aag aag 752 Gln Thr Trp Leu Trp Arg Ser Ala Lys Asp Phe Asn Arg Leu Lys Lys 165 170 175 80 aag aeg cag ect cea gea gcc gea gcc acc ccg falls ccg ggg gct cat 800 Lys Met Gln Pro Pro Ala Ala Ala Val Thr Leu His Leu Gly Ala His 185 190 195 ggc ccc cgacccccga ccctctcccc ctcgctcccc ctccaaaccc cgcecccact 856 Gly Phe ccgcgagagc cagccccgca cgccaacacc tgttgagcca ggagacagaa gccgtgagcc 916 tctggccctt tcctggaccg gctgggcgtg tgatgcgatc agccctgcce cccccccacc 976 tcccaaaggt ctaccgagct ggggaggagg tacagtaggc cctgtcctgc cctgtctcta 1036 caggaagtca tgcccgaggg agtgtgaagt ggttcaggte ggcgcagagg cgcccacggc 1096 ccgcccacca cccccgcctc cttggccagt gcccacccag ccccecaggc ggcacaeceg 1156 gagggcaggg gctgaggggc caccaccaca caegcceecc tggggcgaag ccctttggct 1216 gccccactct ccteggacgg gcgctgcccc cccatcccca aatcactcea cacaeccaat 1276 aacatggtgg tcaggaaaca caatcctaca caaaaagaga tgagatcaac agcgcagggt 1336 tggggecegc attggaggtg ccccacaaac aacactgaa cagaagagaa to gcacaggggc 1396 agggacagac cagaccagac ccaggagtct gagtggcaaa ccaaagcaca caaaacccga 1456 gctgagcatc aggaccttgc ctcgaattgt cteccagtae eacggtgcct cttctctgcc 1516 ccceetccca gggtatctge gggccgccag gceggggagg gcaaccaeag ccacaccaca 1576 ggaectcccg aaagttCaca acgcagtagc attttgggge gcagggtggc agceccccaa 1636 ggcccegccc cccagcccca cccacecaCg accctaageg egCCgcaCea aeaettactc 1696 acctggagat gteacccact agaegatatt taccgcagaa cctccacecc tgtatcaaca 1756 aataaaatgc tCgccccaga aaaa acaaaaaaaa 1790 MLACLCTVLWHLPAVPA / LNRTGDPGPGPSIQKTYDLTRYLEHQLRSLAGTYLNYLGPPFNEPDFNPPRLGAE I TLPRATVDLEVWRSLNDKLRLTQNYEAYSHLLCYLRGLNRQAATAELRRSLAHFCTSLQGLLG? IAGVMAALG YPLPQPLPGTEPT TPGPAHSDFLQKMDDFWLLKELQTWLWRSAKDFNRLKKKMQPPAAAVTLHLGAHGF Table 1 (continued): Rodent, for example, mouse, IL-B60 (SEQ ID NO: 3 and 4): aeg cea gcc cgc cea cgc aeg gcg ccg cgg falls ctc ect gea gtg cea 48 Met Leu Wing Cys Leu Cys Thr Val Leu Trp His Leu Pro Wing Val Pro -15 -10 -5 gct ect aat cgc here gga gac cea ggc ect sgc ccc tec ate cag aaa 96 Wing Leu Asn Arg Thr Gly Asp Pro Gly Pro Gly Pro Be He Gln Lys -1 1 5 10 15 acc tat gac ctc acc cgc falls ccg gag falls ca ccc cgc age cea gcc 144 Thr Tyr Asp Leu Thr Arg Tyr Leu Glu His Gln Leu Arg Ser Leu Wing 20 25 30 ggg acc fall ccg aac Fall ccg ggg ccc ect ccc aac gag ccc gac ccc 192 Gly Thr Tyr Leu Asn Tyr Leu Gly Pro Pro Phe Asn Glu Pro Asp Phe 35 40 '45 aac ccc ccc cga ggg gea gaa acc ccg ccc agg gcc aeg gcc aac 240 Asn Pro Pro Arg Leu Gly Ala Glu Thr Leu Pro Arg Ala Thr Val Asn 50 55 60 ttg gaa gtg tgg cga age ctc aat gac agg ctg cgg ctg acc cag aac 288 Leu Glu Val Trp Arg Ser Leu Asn Asg Arg Leu Arg Leu Thr Gln Asn 65 70 75 tac gag gcg tac age falls ctc ctg cgc falls ccg cgc ggc ccc aac cgc 336 Tyr Glu Wing Tyr Ser His Leu Leu Cys Tyr Leu Arg Gly Leu Asn "Arg 80 85 90 95 cag gcc gcc here gcc gaa ccc cga cgc age ccg gcc falls cec ege acc 384 Gln Ala Ala Thr Ala Glu Leu Arg Arg Ser Leu Ala His Phe Cys Thr 100 105 110 age ctc cag ggc ccg ccg ggc age ate gea ggt gcc atg gcg aeg ccc 432 Ser Leu Gln Gly Leu Leu Gly Ser He Wing Gly Val Mee Wing Thr Leu 115 120 125 ggc cae cec ccg ccc cag ccc ceg ggg gg ace ace gag cea gcc cgg ggg cg ggc ccc gcc ccc ccc ccc cg cg aag aeg gat gac ccc cgg cg cgg 528 Pro Gly Pro Wing His Ser Asp Phe Leu Gln Lys Mee Asp Asp Phe Trp 145 150 155 ccg ctg aag gag ctg cag acc tgg cta tgg cgt tea gcc aag gac tte 576 Leu Leu Lys Glu Leu Gln Thr Trp Leu Trp Arg Ser Wing Lys Asp Phe 160 165 170 175 aac cgg aag aag aag atg cag ect cea gea gct tea gtc acc ctg 624 Asn Arg Leu Lys Lys Lys Mee Gln Pro Pro Wing Wing Val Thr Leu 180, 185 190 ccg gag gcc cat ggt tte tga 648 His Leu Glu Wing His Gly Phe 195 Table 1 (continued): ^^ Z ^^ SLNDRLRLTQIRÍEAYSHLLCYLRGLNRQAATAELRRSLAHFCTSLQGLLGSIAGVMATLG YPLPQPLPGTEPAWAPGPAHSDFLQKMDDFV? LKELQTWLWRSA DFNP KKKMQPPAASVTLHLEAHGF TABLE 2 Comparison of primate and rodent modalities of IL-B60, both the nucleotide and the amino acid sequence. hIL-B60 ATGTTAGCGTGCCTGTGCACGGTGCTCTGGCACCTCCCTGCAGTGCCAGCTCTCAATCGC mIL-B60 ATGTTAGCTTGCCTATGCACGGTGCTGTGGCACCTCCCTGCAGTGCCAGCTCTTAATCGC hIL-B60 ACAGGGGACCCAGGGCCTGGCCCCTCCATCCAGAAAACCTATGACCTCACCCGCTACCTG mi -B60 ACAGGAGATCCAGGCCCTGGCCCCTCCATCCAGAAAACCTATGACCTCACCCGCTACCTG hIL-B60 GAGCACCAACTCCGCAGCTTGGCTGGGACCTATCTGAACTACCTGGGCCCCCCTTTCAAC miL-B60 GAGCATCAACTCCGCAGCTTAGCTGGGACCTACCTGAACTACCTGGGGCCCCCTTTCAAC hIL-B60 GAGCCAGACTTCAACCCTCCCCGCCTGGGGGCAGAGACTCTGCCCAGGGCCACTGTTGAC mIL-B60 GAGCCTGACTTCAATCCTCCTCGACTGGGGGCAGAAACTCTGCCCAGGGCCACGGTCAAC hIL-B60 TTGGAGGTGTGGCGAAGCCTCAATGACAAACTGCGGCTGACCCAGAACTACGAGGCCTAC mIL-B60 TTGGAAGTGTGGCGAAGCCTCAATGACAGGCTGCGGCTGACCCAGAACTATGAGGCGTAC hIL-B60 AGCCACCTTCTGTGTTACTTGCGTGGCCTCAACCGTCAGGCTGCCACTGCTGAGCTGCGC mIL-B60 AGTCACCTCCTGTGTTACTTGCGTGGCCTCAACCGTCAGGCTGCCACAGCTGAACTCCGA hIL-360 CGCAGCCTGGCCCACTTCTGCACCAGCCTCCAGGGCCTGCTGGGCAGCATTGCGGGCGTC mIL-B60 CGTAGCCTGGCCCACTTCTGTACCAGCCTCCAGGGCCTGCTGGGCAGCATTGCAGGTGTC hIL-B60 ATGGCAGCTCTGGGCTACCCACTGCCCCAGCCGCTGCCTGGGACTGAACCCACTTGGACT mIL-B60 ATGGCGACGCTTGGCTACCCACTGCCCCAGCCTCTGCCAGGGACTGAGCCAGCCTGGGCC hIL-B60 CCTGGCCCTGCCCACAGTGACTTCCTCCAGAAGATGGACGACTTCTGGCTGCTGAAGGAG mIL-B60 CCTGGCCCTGCCCACAGTGACTTCCTCCAGAAGATGGATGACTTCTGGCTGCTGAAGGAG Yes? íS Table 2 (continued): Alignment of IL-B60: the underlines are proposed helices. In general, those residues that are in helix A and D and do not point inward towards the center (mostly hydrophobic residues in helix A and D) are the most likely residues to interact with receptors.

A hIL-B60 MLACLCTVLWHLPAVPALNRTGDPGPGP? IOKTYDLTRYLEHOLRSLAGT mIL-B60 MLACLCTVLWHLPAVPALNRTGDPGPGPSIOKTYDLTRYLEHOLRSLAGT 15B Í1IL-B60-B60 XLNYLGPPFNEPDFNPPRLGAETLPRATVDLEV RSLNDKLRLTONYEAY mIL YLN GPPFNEPDFNPPPXGAETLPRATVNLEVWRSLNDRLRLTONYEAY C ^ hIL-B60 SHLLCYLRGLNROAATAELRRS AHFCtS Op GSIAGVMAALGYPLPO mIL-B60 SHLLCYLRGLNROAATAELRRSLAHF TSLOG GSIAGVMATLGYPL, PO hIL-B60 PPAAAVTLKLGAHGF mIL-B60 PPAASVTLHLEAHGF TABLE 3 Comparison of several cytokines compared with IL-B60. Human IL-B60 is SEQ ID NO: 2; Mouse IL-B60 is SEQ ID NO: 4; Mouse LIF (mLlF) is SEQ ID NO: 5 and Access number X06381; Human LIF (hLIF) is SEQ ID NO: 6 and Access numbers M63420 J05436; Human CT-1 (hCT-1) is SEQ ID NO: 7 and Access number U43030; Mouse CT-1 (mCT-1) is SEQ ID NO: 8 and Access number U18366; Human CNTF (hCNTF) is SEQ ID NO: 9 and Access number X60542; Mouse CNTF (mCNTF) is SEQ ID NO: 10 and access number U05342- DNAX human IL-40 (hDIL-40) is SEQ ID NO: 11. mLlF -M VLAAGIVPLLLLVLHWKHGAGSPLPI -TPVNATC-AIRHPCHGNLMN hLIF -M VLAAGWP -LLLVLHWKHGAGSPLPI - TPVNATC-AIRHPCHNNLMN hCT-1 - -MSRREGSLE- - -D-- PQTDSSVSLLPH-LEA KIRQT-HS - LA mCT-1 - -MSQREGSLE-- -D- -HQTDSSISFLPH-LEA KIRQT-HN- -LA hIL-360 -MLACLCTVLW HLPAVPALNRTG-DPG-PGP-SIQKT-YD - LT mIL-B60 -MLACLCTVLW HLPAVPALNRTG-DPG-PGP-SIQKT-YD - LT hCNTF MAFTE HSPLTPHR-R --- DL-CSR-SIW THE mCNTF MAFA? QSPLTLHR-R D-L-CSR-SIW LA hDIL-40 MTHLSLLGPLPCVRTSQQLPETQQVTTPGKKPVSVGRREVRVP GT mLlF QIKNQLAQLNGSA ALFISYYTAQGEPF - PNNVEK-LCAPNMTDFPSFH hLIF QIRSQLAQLNGSANALFILYYTAQGEPF - PNNLDK-LCGPNVTDFPPFH hCT-1 HLLT YAEQ LLQEYVQLQGDPFGLPSFSPPRLPVAGLSAPAPSH mCT-1 RLLTKYAEQ LLEEYVQQQGEPFGLPGFSPPRLPLAGLSGPAPSH hIL-B60-B60 RYLEHQLRS LAGTYLNYLGPPFNEPDFNPPRLGAETLPRATVDL mIL RYLEHQLRS LAGTYLNYLGPPFNEPDFNPPRLGAETLPRATVNL. hCNTF RKIRSDLTA LTESYVKHQG - LNK NINLDSADGMPVASTD- mCNTF RKIRSDLTA LMESYVKHQG - LNK NISLDSVDGVPVASTD- hDIL-40 ALVPSLLSV SVLLQLQYQGSPFSDPGF? APELQLSSLPPATAFF LIF GNGTEKTKLVELYRMVAYLSASLTNITR-DQKVLNPTAVSLQVKLNA hLIF ANGTEKAKLVELYRIWY DQKILNPSALSLHSKLNA GTSLGNITR-hCT-1-RQAELNPRAPRLLRRLED AGLPVHERLRLDAAALAALPPLLDAVCR mCT-1-RQAELNPRAPRLLRSLED AGLPVSERLRQDAAALSVLPALLDAVRR hIL-B60 EVWRSLNDKLRLTQNYEAYSHLLCYLRGLN - RQAATAELRRSLAHFCTS mIL-B60-RQAATAELRRSLAHFCTS EVWRSLNDRLRLTQNYEAYSHLLCYLRGLN hCNTF -QWSELTEAERLQENLQAYRTFHVLLARLLEDQQVHFTPTEGDFHQAIHT mCNTF -RWSEMTEAERLQENLQAYRTFQGMLTKLLEDQRVHFTPTEGDFHQAIHT HDIL-40 KTWKALDDG RLSLAQRAID PHLQLVED-DQSDLNPGSPILPAQLGA mLIF TIDVMRGLLSNVLCRLCNKYRV - GHVDVPP VPDHSDKE - AFQR hLIF TADILRGLLSNVLCRLCSKYHV - GHVDVTY GPDTSGKD - VFQK hCT-1 AA QARALGAAVEALLAALGAANRGPRAEPP - AATASAASATG - VFPA mCT-1 AARQVRALGAAVETVLAALGAAARGPGPEPVTVATLFTANSTAG-- IFSA hIL-B60 LQGLLGSIAGVMAALGYPLPQP - LPGTEPT WTPGPAHS DFLQ mIL-B60 LQGLLGSIAGVMATLGYPLPQP - LPGTEPA WAPGPAHS DFLQ hCMTF LLLQVAAFAYQIEELMILLEYK - IPRNEAD GMPINVGDGG-LFEK mCNTF LTLQVSAFAYQLEELMALLEQK - VPEKEAD GMPVTIGDGG-LFEK HDIL-40 ARLRAQGPLGNMAAIMTALGLP - IP-SDEs TPGLAAFGASAFER Table 3: (continued): mLIF KKLGCQLLGTYKQVIS WQAF hL F KKLGCQLLGKYKQIIA VLAQAF hCT-1 KVLGLRVCGLYREWLSRTEGDLGQLLPGGSA- mCT-1 KVLGFHVCGLYGEWVSRTEGDLGQLVPGGVA- hIL-B60 -B60 KMDDFWLLKELQTWLWRSAKDFNRLKKKMQPPAAAVTLHLGAHGF-- ml KMDDFWLLKELQTWLWRSAKDFNRLKKKMQPPAASVTLHLEAHGF-- hCNTF KLWGLKVLQELSQWTVRSIHDL-RFI SHQTGIPARGSHYI.AN KKM mCNTF KLWGLKVLQELSQWTVRSIHDL-RVIS HHMGISAHESHYGA -? KQM HDIL-40 KCRGYWTREYGHWTDRAVRDLALLKAKYSA The structural homology of IL-B60 with related cytokine proteins suggests the related function of this molecule. IL-B60 is a long-chain cytokine that exhibits sequence similarity to IL-6 and G-CSF. IL-B60 agonists, or antagonists, can also act as receptor or functional antagonists, for example, blocking the binding of G-CSF or IL-6 to their respective receptors, or acting as intermediaries in the opposite actions. Thus, IL-B60, or its antagonists, may be useful in the treatment of abnormal medical conditions, including immune disorders, for example, immune deficiencies of T cells, chronic inflammation, or rejection of tissues, or in cardiovascular or neurophysiological conditions. Natural antigens are capable of mediating various biochemical responses that lead to biological or physiological responses in target cells. The preferred embodiment characterized herein is human, but other primate counterparts, or other species, exist in nature. Additional sequences for proteins should also be available in other mammalian species, for example, primates, canines, felines and rodents. See below. The descriptions below are directed, for exemplification purposes, to a human IL-B60, but are applicable in the same way to related modalities of other species. In particular, the association of IL-B60 with a partner has been confirmed. The molecules IL-B60 and CLF-1 have probably evolved together, reflected in their homology between species. For example, the co-evolution of its function is suggested by the observation that the human / mouse relationship of IL-B60 is very close, as is human / mouse CLF-1. If the two are combined functionally, they could act together in the manner of IL-12. See, for example, Trinchieri (1998) Adv. Immunol. 70: 83-243; Gately et al. (1998) Ann. Rev. Immunol. 16: 495-521; and Trinchieri (1998) Int. Rev. Immunol. 16: 365-396.

However, as a complex, they will interact with two high receptors in the family of cytokine receptors, for example, gp130, LIF-R, OSM-R, IL-12RM, IL-12Rb2 and NR30. These receptors will be evaluated to determine the link to the soluble complex. A series of BAF / 3 cells that stably express several of these high receptors has been constructed. The supernatants of transfectants of both IL-B60 and CLF-1 (or a unique combination construct) in the same cell will be used to evaluate these various BAF / 3 cells to see if there is a proliferative or other signal response. As such, most of the physiological effects of the cytokine may be due to the protein complex. As such, many of the descriptions below of the biology resulting from the cytokine can actually be effected physiologically by the complex comprising the combination of the subunits. Table 4 provides the sequences of the IL-B60 partner, known as CLF-1. The CNTF receptor (CNTF-R) alpha subunit was described, for example, by Davis et al. (1991) Science 253: 59-63. See also access numbers GenBank NM1001842 and M73238 (human); AF068615 (mouse); and S54212 (rat); each of which is incorporated herein by reference.

TABLE 4 Factor 1 Alignment of Mouse and Human Type Cytokine (CLF-1; SEQ ID NO: 12 and 13). See Elson et al. (1998) J. Immunol. 161: 1371-1379; GenBank access number AFO59293 and NM_004750; also described by Douglas J. Hilton (Australia) in W09920755. The signal sequence reported of 37 amino acids in human form, decomposition in GSG 1 AHT. hCLF- l MPAGRRGPAAQSARRPPPLLPLLLLLCVLGAPRAGSGAHTAVISPQDPTL mCLF- 1 RPLSSLWSPLLLCVLGVPRGGSGAHTAVISPQDPTL hCLF-l LIGSSLLATCSVKGDPPGATAEGLYWTLNGRRLPPELSRVLNASTLALAL mCLF-1 LIGSSLQATCSIHGDTPGATAEGLYWTLNGRRLP-SLSRLLNTSTLALAL hCLF- l ANLNGSRQRSGDNLVCHARDGS ILAGSCL-YVGLPPEKPT? JI SCWSKNMKD mCLF- 1 ANLNGSRQQSGDNLVCHARDGS ILAGSCLYVGLPPEKPFNISCWSRNMKD hCLF- l LTCRWTPGAHGETFLHTNYSLKYKLRWYGQDNTCEEYHTVGPHSCHI PKD mCLF- 1 LTCRWTPGA-HGETFLHTNY? LKYKLRWYGQDNTCEEYHTVGPHSCHI PKD hCLF- l LALFTPYEIWVEATNRLGSARSDVLTLDILDWTTDPPPDVHVSRVGGLE mCLF- 1 LALFTPYEIWVEATMRLGSARSDVLTLDVLDV ^ TTDPPPDVHVSRVGGL? hCLF- 1 DQLSVR S PPALKDFLFQAKYQIRYRVEDSVDWKVVDDVSNQTSCRLAG mCLF- 1 DQLSVRWVS PPALKDFLFQAKYQIRYRVEDSVDWKWDDVSNQTSCRLAG hCLF-l LKPGTVYFVQVRCNPFGIYGSKKAGIWSEWSHPTAASTPRSERPGPGGGA mCLF-1 LKPGTVYFVQVRCNPFGIYG? KKAGIWSEWSHPTAASTPRSERPGPGGGV 3 hCLF- l CEPRGGEPSSGPVRRELKQFLGWLKKHAYC? NLSFRLYDQWRAWMQKSHK mCLF- 1 CEPRGGEPSSGPVP.RELKQFLGWLKKKAYC? NLSFRLYDQWRAWMQKSHK hCLF- l TRNQ VLPDKL mCLF- 1 TRNQDEGILPSGRRGAARGPAG The standard domains of the human CLF-1 receptor sequence correspond approximately to: signal from 1 to approximately 38; first domain of type IG from about residue 39 to 130; a second domain from about 131 to about 237; and the last from about 238 to the end. The descriptions below can also be applied to CLF-1, or to the IL-B60 / CLF-1 complex. A fusion of IL-B60 with CLF-1 can be constructed, such as, for example, hyper IL-6. See, for example, Flscher et al. (1997) Nature Biotechnol. 15: 142-145; Rakemann et al. (1999) J, Biol. Chem. 274: 1257-1266; and Peters et al. (1998) J. Immunol. 161: 3575-3581; which are incorporated herein by reference. The original discovery and molecular characterization of CNTF as a potent survival factor for neuronal cells (see for example, Hughes et al. (1988) Nature 335: 70-73; and Stockii et al. (1989) Nature 342-, 920-923) suggested an alleged therapeutic use as a molecule that could accelerate the repair of divided or damaged motor neurons (Sendtner et al. (1990) Nature 345: 440-441; and Curtis et al. (1993) Nature 365: 253-256) or prevent nerve degeneration (Sendtner et al. (1992) Nature 358: 502-504; Emerich et al. (1997) Nature 386: 395-399; and Gravel et al. (1997) Nature Med. 3: 765-770). However, CNTF is strangely a protein without a secretory signal peptide, and does not seem to escape from the cell (Stockii et al., Ibid); In addition, the designed interruptions (Masu et al. (1993) Nature 365: 27-32) or natural interruptions (Takahashi et al. (1994) Nature Genet. 7: 79-84) of the CNTF gene are not harmful. In contrast, interruptions of the primary receptor gene for CNTF (CNTF-Ra) prove lethal shortly after birth. DeChiara et al. (1995) Cell 83: 313-322). Together, these observations point to the existence of a second ligand for CNTF-Ra that is physiologically responsible for the observed in vitro, or in vivo, actions of CNTF desired. This work shows that the compound cytokine IL-B60 / CLF-1 is probably this long-sought factor, which is so decisive from the point of view of development, it is secreted from target organs and directs its innervation by motor neurons, as well as also promising from the therapeutic point of view, since the nervous transaction produces a rapid and lasting induction of both IL-B60 and CLF-1, indicating a function for the complex in regeneration. In support of this model, the genetic disruption of CLF-1 (Alexander and others (1999) Curr. Biol. 9: 605-608) is very similar in phenotype to knockout of CNTF-Ra. In an intriguing peculiarity, although IL-B60 has a signal peptide, its secretion remains critically dependent on the complex with CLF-1, as described. Once secreted, IL-B60 signals by means of a tripartite receptor system that it is otherwise identical to that of CNTF, consisting of the two signal transducer components expressed in ubiquitous gp130 and LIF-R, and the receptor determinant of the specificity, CNTF-Ra. Surprisingly, the function of CLF-1 seems to be restricted to that of a chaperone, since it is discarded from the signaling complex after supplying IL-B60 to CNTF-Ra; in fact, the requirement for CLF-1 can be set aside by fusing IL-B60 directly to a soluble form of the CNTF-Ra chain. The three protein chains involved in this new system, IL-B60, CLF-1 and CNTF-Ra, represent the most highly conserved sequences in the hematopoietic / receptor cytokine superfamily, indicating a decisive interaction from the point of view of evolution. In addition, the conditional use of a hematopoietic receptor as a secretion and escort factor presents a new paradigm for cytokine activity. To summarize, this work illuminates the interlaced biological function of two orphan molecules, IL-B60 and CLF-1, describing their new coupling of the CNTF receptor complex. In doing so, we present a strong argument, in that it is the cytokine IL-B60 / CLF-1 that serves as a key physiological factor in the development and regeneration of motor neurons.

II. Purified IL-B60 or complex The human amino acid sequence IL-B60 is displayed, in a form within SEQ ID NO: 2. Other natural nucleic acids encoding the protein can be isolated by standard procedures using the sequence provided, for example, PCR techniques, or by hybridization. These amino acid sequences, so long as they are amino to carboxy, are important to provide sequence information for the cytokine, allowing the distinction of protein antigen, other proteins, and exemplifying numerous variants. Moreover, the peptide sequences allow the preparation of peptides to generate antibodies to recognize said segments, and the nucleotide sequences allow the preparation of oligonucleotide probes, which are both strategies for the detection or isolation, for example, cloning of genes that encode these sequences. As used herein, the term "human soluble IL-B60" will encompass, when used in a protein context, a protein having an amino acid sequence corresponding to a soluble polypeptide of SEQ ID NO: 2. Significant fragments of the they will often retain similar functions, for example, antigenicity. Preferred embodiments comprise a plurality of distinctive segments, for example, that do not overlap, of the specified length. Typically, the plurality will be at least two, more usually at least three, and preferably 5, 7 or even more. While the minimum length can be quoted, longer lengths of various sizes may be appropriate, for example, one of length 7, and two of length 12. Similar characteristics to polynucleotides apply. Linker components, e.g., antibodies, typically bind to an IL-B60 with high affinity, eg, at least about 100 nM, usually better than about 30 nM, preferably better than about 10 nM, and with more preference to better than about 3 nM. The counterpart proteins will be found in mammalian species that are not human, for example, other primates, ungulates or rodents. Non-mammalian species should also possess structurally or functionally related genes and proteins, for example, birds or amphibians. The term "polypeptide" as used herein includes a segment or significant fragment, and encompasses a stretch of amino acid residues of at least about 8 amino acids, generally at least about 12 amino acids, typically at least about 16 amino acids, preferably at least about 20 amino acids, and in particularly preferred embodiments, at least about 30 or more amino acids, for example 35, 40, 45, 50, etc. Said fragments may have ends that begin and / or end up in practically all positions, for example, beginning at residues 1, 2, 3, etc., and ending at, for example, 150, 149, 148, etc., in all practical combinations. Particularly interesting peptides possess ends corresponding to the structural domain boundaries, for example, helices A, B, C and / or D. See Table 1. The term "binding composition" refers to molecules that bind with specificity to IL-B60, for example, in an antibody-antigen interaction. The specificity can be more or less inclusive, for example, specific to a particular modality, or to groups of related modalities, for example, primate, rodent, etc. Depletion or absorptions can provide desired selectivities. In addition there are provided compounds, for example, proteins, which specifically associate with IL-B60, including in a natural physiologically relevant protein-protein interaction, either covalent or non-covalent. The molecule can be a polymer, or chemical reagent. A functional analog can be a protein with structural modifications, or it can be a molecule having a molecular form that interacts with the appropriate binding determinants. The compounds can serve as agonists or antagonists of a receptor binding interaction, see, for example, Goodman et al. (Eds.) Goodman &; Gilman's: The Pharmacoloqical Bases of Therapeutics (current ed.) Pergamon Press. Substantially pure, for example, in a protein context, typically means that the protein is free of other contaminating proteins, nucleic acids, or other biological derivatives of the original source organism. The purity can be tested by standard methods, typically by weight, and will commonly be at least about 40% pure, generally at least about 50% pure, often at least about 60% pure, typically at least about 80% pure. % pure, preferably at least about 90% pure, and in the most preferred embodiments, at least about 95% pure. Often carriers or excipients will be added. The solubility of a polypeptide or fragment depends on the environment and the polypeptide. Many parameters affect the solubility of the polypeptide, including temperature, electrolyte environment, size and molecular characteristics of the polypeptide and nature of the solvent. Typically, the temperature at which the polypeptide is used ranges from about 4 ° C to about 65 ° C. Usually the temperature in use is greater than about 18 ° C. For diagnostic purposes, the temperature will usually be about room temperature or warmer, but lower than the denaturing temperature of the components in the assay. For therapeutic purposes, the temperature will usually be body temperature, typically about 37 ° C for humans and mice, although under certain situations the temperature may rise or fall in situ or in vitro. The size and structure of the polypeptide in general should be in a substantially stable state, and usually not in a denatured state. The polypeptide can be associated with other polypeptides in a quaternary structure, for example, to confer solubility, or it can be associated with lipids or detergents. The solvent and the electrolytes will usually be a biologically compatible pH regulator, of a type used for the preservation of biological activities, and will usually approximate a physiological aqueous solvent. Usually, the solvent will have a neutral pH, typically between about 5 and 10, and preferably about 7.5. In some instances, one or more detergents will be added, typically a non-denaturing mild detergent, for example, CHS (cholesterol hemisuccinate) or CHAPS (3- [3-colamidopropyl) dimethylammoniol-1-propanesulfonate), or a sufficiently low concentration in a manner to avoid significant alteration of the structural or physiological properties of the protein. In other cases, a harsh detergent may be used to effect significant denaturation. Typically, an IL-B60 polypeptide that specifically binds, or that is specifically immunoreactive with, an antibody, for example, such as a polyclonal antibody, generated against a defined immunogen, for example, such as an immunogen that is determined in an immunoassay, is determined. consists of an amino acid sequence of SEQ ID NO: 2 or fragments thereof, or a polypeptide generated from the nucleic acid of SEQ ID NO: 1. Included within the limits of the present invention are those nucleic acid sequences that are describe herein, including functional variants, that encode polypeptides that selectively bind to polyclonal antibodies raised against the prototypical IL-B60 polypeptide as structurally and functionally defined herein. The immunoassay typically utilizes a polyclonal antiserum that was produced, for example, for a protein of SEQ ID NO: 2. This antiserum is selected, or subjected to depletion, in a manner to have low cross-reactivity against other closely related family members. , preferably of the same species, and any such cross-reactivity is eliminated by immunoabsorption or depletion before use in the immunoassay. Appropriate selective serum preparations can be isolated and characterized. To produce antisera for use in an immunoassay, the protein, for example, of SEQ ID NO: 2, is isolated as described herein. For example, recombinant protein can be produced in a mammalian cell line. A suitable host, for example, an inbred strain of mice, such as Balb / c, is immunized with the protein of SEQ ID NO: 2, using a standard adjuvant, such as Freund's adjuvant, and a standard mouse immunization protocol (see Harlow and Lane). Alternatively, a synthetic peptide of substantially complete length derived from the sequences described herein, can be used as an immunogen. The polyclonal sera are collected and titrated against the immunogenic protein in an immunoassay, for example, a solid phase immunoassay with the immunogen immobilized on a solid support. Police antisera with a titer of 104 or more are selected and evaluated to determine their cross-reactivity against other closely related family members, for example, LIF, CT-1, CNTF, DIL-40, or other family members. IL-6, using a competitive binding immunoassay, such as that described in Harlow and Lane, supra, at pages 570-573. Preferably, at least two members of the IL-6 / IL-12 family they are used in this determination, in conjunction with the objective. These members of the long chain cytokine family can be produced as recombinant proteins and isolated using standard molecular chemistry and protein chemistry techniques, as described herein. Thus, antibody preparations having desired selectivity or specificity can be identified or produced for subclasses of members of the IL-B60 family. Alternatively, antibodies that bind to the complex comprising IL-B60 with CLF-1 can be prepared. Immunoassays can be used in the competitive binding format for cross-reactivity determinations. For example, the protein of SEQ ID NO: 2 can be immobilized to a solid support. The proteins added to the assay compete with the binding of the antisera to the immobilized antigen. The ability of the proteins mentioned above to compete with the binding of the antisera to the immobilized protein is compared to the protein of SEQ ID NO: 2. The percent cross-reactivity for the above proteins is calculated using standard calculations. Those antisera with less than 10% cross-reactivity with each of the proteins mentioned above are selected and pooled. The cross-reactive antibodies are then removed from the pooled antisera by immunoabsorption with the proteins mentioned above. The immunosorbed and pooled antisera are then used in a competitive binding immunoassay as described above, to compare a second protein to the immunogen protein. To make this comparison, the two proteins are each tested at a wide range of concentrations, and the amount of each protein required to inhibit 50% of the binding of the antisera to the immobilized protein is determined. If the amount of the second protein required is less than two times the amount of the protein of, eg, SEQ ID NO: 2 that is required, then the second protein is said to bind specifically to an antibody generated for the immunogen.

III. Physical variants This invention also encompasses proteins or peptides having substantial amino acid sequence identity, with the amino acid sequence of the 11-B60 antigen. Variants include allelic, polymorphic or species variants. The homology of amino acid sequence, or identity of sequences, is determined by optimizing the confrontation of residues, if necessary, introducing spaces as required. See also Needleham et al. (1970) J. Mol. Biol. 48: 443-453; Sankoff and others (1983) Chapter One in Time Warps, String Edits. and Macromolecules: The Theorv and Practice of Sequence Comparison, Addison-Wesley, Reading, MA; and software packages from IntelliGenetics, Mountain View, CA; and the University of Wisconsin Genetics Computer Group, Madison, Wl. The identity of sequences changes when conservative substitutions are considered as confrontations. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid; asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. Conservation can be applied to biological characteristics, functional characteristics, or structural characteristics. Homologous amino acid sequences are typically intended to include variations between species and allelics or natural polymorphs of a protein sequence. Peptides or typical homologous proteins will have from 25-100% identity (if spaces can be introduced), at 50-100% identity (if conservative substitutions are included) with the amino acid sequence of IL-B60. The identity measurements will be at least about 35%, generally at least about 40%, often at least about 50% typically at least about 60%, usually at least about 70%, preferably at least about 80%, and more preferably at least about 90%. The isolated IL-B60 DNA can be modified without difficulty by nucleotide substitutions, nucleotide deletions, nucleotide insertions and inversions of short nucleotide stretches. These modifications produce new DNA sequences encoding these antigens, their derivatives, or proteins that have similar physiological, immunogenic, antigenic or other functional activity. These modified sequences can be used to produce mutant antigens or to improve expression. Improved expression may involve genetic amplification, increased transcription, increased translation, and other mechanisms. "Mutant IL-B60" encompasses a polypeptide that is otherwise within the definition of sequence identity of IL-B60 as set forth above, but which has an amino acid sequence that differs from that of IL-B60 as normally found in nature, either by way of deletion, substitution, or insertion. This generally includes proteins that have significant identity with a protein having sequence of SEQ ID NO: 2, and that share several biological activities, eg, antigenic or immunogenic, with those sequences, and in preferred embodiments contain most of the described sequences full length natural. Full-length sequences will typically be preferred, although truncated versions will also be useful, similarly, genes or proteins found in natural sources are typically the most desired. Similar concepts apply to different IL-B60 proteins, particularly those found in several warm-blooded animals, e.g., mammals and birds. These descriptions are generally intended to encompass all IL-B60 proteins, not limited to the particular primate modalities that are specifically discussed. The mutagenesis of IL-B60 can also be conducted by making insertions or deletions of amino acids. The substitutions, deletions, insertions or any combination can be generated to arrive at a final construction. Inserts include amino- or carboxy-terminal fusions. Random mutagenesis can be conducted in an objective codon, and the expressed mutants can then be screened to determine the desired activity. Methods for making substitution mutations at predetermined sites in DNA having a known sequence are well known in the art, for example, by mutagenesis of M13 primer or polymerase chain reaction (PCR) techniques. See, for example, Sambrook et al. (1989); Ausubel et al. (1987 and Supplements); and Kunkel et al. (1987) Methods in Enzvmol. 154: 367-382. Preferred embodiments include, for example, 1-time, 2-fold, 3-fold, 5-fold, 7-fold, etc., preferably conservative substitutions at the nucleotide or amino acid levels. Preferably the substitutions will be outside the conserved cisterns, and will often be in the regions outside the helical structural domains. Such variants may be useful for producing specific antibodies, and will often share many or all of the biological properties. See Table 2. The recognition of the cytokine structure provides important insights into the structure and positions of the residues that can be modified to effect desired changes in receptor interaction. In addition, the interaction of IL-B60 with the CLF-1 protein requires complementary structural features at the interaction surface. The present invention also provides recombinant proteins, for example, heterologous fusion proteins using segments of these proteins. A heterologous fusion protein is a fusion of proteins or segments that naturally are not normally fused in the same way. A similar concept applies to heterologous nucleic acid sequences. In addition, new constructions can be made from the combination of similar functional domains of other proteins. For example, target link segments or other segments can be "exchanged" between different new fragments or fusion polypeptides. See for example, Cunningham et al. (1989) Science 243: 1330-1336; and O 'Dowd et al. (1988) J. Biol. Chem. 263: 15985-15992. The phosphoramidite method described by Beaucage and Carruthers (1981) Tetra. Letts. 22: 1859-1862, will produce suitable synthetic DNA fragments. A double stranded fragment will often be obtained, either by synthesizing the complementary strand and by annealing the strand, together, under appropriate conditions, or by adding the complementary strand using DNA polymerase with an appropriate primer sequence, eg, PCR techniques.

Structural analysis can be applied to this gene, compared to the IL-6 family of cytokines. The family includes, for example IL-6, IL-11, IL-12, G-CSF, LIF, OSM, CNTF and Ob. The alignment of human and mouse IL-B60 sequences with other members of the IL-6 family should allow the definition of structural features. In particular, ß-sheet and a-helix residues can be determined using, for example, the RASMOL program, see Bazan et al. (1996) Nature 379, 591-, Lodi et al. (1994) Science 263: 1762-1766; Sayle and Milner-White (1995) TIBS 20: 374-37; and Gronenberg et al. (1991) Protein Enqineerinq 4: 263-269. See, also, Wiikins et al. (eds. 1997) Proteome Research: New Frontiers in Functional Genomics Springer-Verlag, NY. The preferred residues for substitutions include surface exposed residues that would be predicted to interact with the receptor. Other residues that should retain the function will be conservative substitutions, in particular in a position far from the surface exposed residues.

IV. Functional variants Blocking the physiological response to IL-B60 may result from competitive inhibition of ligand binding to its receptor. The in vitro assays of the present invention will often use isolated protein, soluble fragments comprising receptor binding segments of these proteins, or fragments attached to solid phase substrates. These assays will also allow for diagnostic determination of the effects of either linkage segment mutations and modifications, or cytokine mutations and modifications, eg, IL-B60 analogues. This invention further contemplates the use of competitive drug screening assays, for example, when neutralizing antibodies to the cytokine, or receptor binding fragments compete with an evaluating compound. "Derivatives" of the IL-B60 antigens include mutants of naturally occurring amino acid sequences, glycosylation variants and covalent conjugates or of aggregates with other chemical moieties. Covalent derivatives can be prepared by linking functionalities to groups that are in amino acid side chains of IL-B60, or in the N or C terms, for example, by standard means. See, for example, Lundblad and Noyes (1988) Chemical Reaqents for Protein Modification. vol. 1 - 2, CRC Press, Inc., Boca Raton, FL; Hugli (ed. 1989) Techniques in Protein Chemistry, Academie Press, San Diego, CA; and Wong (1991) Chemistrv of Protein Coniuqatíon and Cross Linkinq. CRC Press, Boca Raton, FL. In particular, glycosylation alterations are included, for example, made by modifying the glycosylation patterns of a polypeptide during its synthesis and processing, or in other processing steps. See, for example, Elbein (1987) Ann. Rev. Biochem. 56: 497-534. In addition, versions of the peptides with the same primary amino acid sequence are contemplated, which have other minor modifications, including phosphorylated amino acid residues, for example, phosphotyrosine, phosphoserine, or phosphothreonine. Fusion polypeptides between IL-B60 and other homologous or heterologous proteins are also provided. Many cytokine receptors or other surface proteins are multimeric, for example, homodimeric entities, and a repeat construct can have several advantages, including decreased susceptibility to proteolytic decomposition. Typical examples are fusions of an informant polypeptide, eg, luciferase, with a segment or domain of a protein, eg, a receptor binding segment, so that the presence or location of the fused ligand can be easily determined. See, for example, Dull et al., U.S. Patent No. 4,859,609. Other genetic fusion partners include bacterial β-galactosidase, trpE, Protein A, β-lactamase, alpha amylase, alcohol dehydrogenase, yeast alpha mating factor and detection or purification tails such as FLAG sequence or His6 sequence. See, for example, Godowski et al. (1988) Science 241: 812-816. Fusion peptides will typically be made either by recombinant nucleic acid methods, or by synthetic polypeptide methods. Techniques for nucleic acid manipulation and expression are generally described, for example in Sambrook, et al. (1989) Molecular Cloning: A Laboratorv Manual (2nd ed.), Vol. 1 - 3, Cold Spring Harbor Laboratory; and Ausubel et al. (eds. 1993) Current Protocols in Molecular Bioioqy, Greene and Wiley, NY. Techniques for the synthesis of polypeptides are described, for example, in Merrifield (1963) J. Amer. Chem. Soc. 85: 2149-2156-, Merrifiedi (1986) Science 232-341-347; Atherton et al. (1989) Solid Phase Peptide-Svnthesis: A Practical Approach, IRL Press, Oxford; and Grant (1992) Svnthetic Peptides: A User's Guide. W. H. Freeman, NY. The folding methods can be applicable to synthetic proteins. This invention also contemplates the use of IL-B60 protein derivatives, other than variations in amino acid sequence or glycosylation. Said derivatives may involve covalent or aggregation association with chemical portions or protein carriers. The covalent or aggregation derivatives will be useful as immunogens, as reagents in immunoassays, or in purification methods such as for affinity purification of binding partners, eg, other antigens. An IL-B60 can be immobilized by covalent binding to a solid support, such as SEPHAROSE, activated with cyanogen bromide by methods that are well known in the art, or adsorbed onto polyolefin surfaces, with or without glutaraldehyde crosslinking, for use in the assay or purification of anti-IL-B60 antibodies, or an alternative binding composition. The IL-B60 proteins can also be labeled with a detectable group, for example, for use in diagnostic assays. The purification of IL-B60 can be effected by an immobilized antibody or complementary binding partner, for example, binding portion of a receptor.

A solubilized IL-B60 or fragment of this invention can be used as an immunogen for the production of antisera or antibodies specific for binding. The purified antigen can be used to screen monoclonal antibodies or antigen binding fragments, encompassing antigen binding fragments of natural antibodies, eg, Fab, Fab ', F (ab) 2, etc. Purified IL-B60 antigens can also be used as a reagent to detect antibodies generated in response to the presence of elevated levels of the cytokine, which can be diagnostic of an abnormal or specific physiological or disease condition. This invention contemplates antibodies produced against amino acid sequences encoded by nucleotide sequence set forth in SEQ ID NO: 1, or fragments of proteins containing it. In particular, this invention contemplates antibodies that have binding affinity to, or that occur against, specific domains, eg, helices A, B, C, or D of IL-B60, or the Ig domains of CLF-1. The present invention contemplates the isolation of variants of additional closely related species. Southern and Northern blot analysis will establish that similar genetic entities exist in other mammals. It is possible that IL-B60 are widely disseminated in species variants, for example, rodents, lagomorphs, carnivores, artiodactyls, perissodactyls, and primates. The invention further provides means for isolating a group of related antigens that exhibit both differences and similarities in structure, expression and function. The elucidation of many of the physiological effects of molecules will be greatly accelerated by the isolation and characterization of additional distinct species or polymorphic variants of them. In particular, the present invention provides useful probes for the identification of additional homologous genetic entities in different species. The isolated genes will allow the transformation of cells lacking the expression of an IL-B60, for example, either species or cell types lacking the corresponding proteins and exhibiting negative background activity. This should allow the analysis of the function of IL-B60 compared to untransformed control cells. The dissection of critical structural elements that affect the various physiological functions mediated through these antigens is possible using standard techniques of modern molecular biology, particularly in the comparison of members of the related class. See, for example, the homologous screening mutagenesis technique described in Cunningham et al. (1989) Science 243: 1339-1336; and approaches used in O'Dowd et al. (1988) J. Biol. Chem. 263: 15985-15992; and Lechleiter et al. (1990) EMBO J. 9: 4381-44090. Intracellular functions would probably involve receptor signaling. However, protein internalization may occur under certain circumstances, and the interaction between intracellular components and cytokine may occur. Specific segments of interaction of IL-B60 with interaction components can be identified by mutagenesis or direct biochemical means, for example, cross-linking or affinity methods. Structural analysis by crystallographic or other physical methods will also be applicable. Further investigation of the mechanism of signal transduction will include the study of associated components that can be isolated by affinity methods or by genetic means, eg, mutant complementation analysis. An additional study of the expression and control of IL-B60 will be sought. The control elements associated with the antigens should exhibit physiological, developmental, tissue-specific, or other expression, differential patterns. The genetic regions towards the 5 'end or towards the 3' end, for example, control elements, are of interest. Structural studies of IL-B60 antigens will lead to the design of new antigens, in particular analogues that exhibit agonist or antagonist properties on the molecule. This can be combined with previously described screening methods to isolate antigens that exhibit desired spectra of activities.

V. Antibodies Antibodies can be produced for several epitopes of the IL-B60 proteins, including allelic, polymorphic or species variants, and fragments thereof, both in their natural forms and in their recombinant forms. Additionally, antibodies to IL-B60 can be produced, either in their active forms or in their inactive forms, including natural or denatured versions. Anti-idiotypic antibodies are also contemplated. Antibodies, including binding fragments and single chain versions, against predetermined fragments of the antigens, can be produced by immunization of animals with conjugates of the fragments with immunogenic proteins. The monoclonal antibodies are prepared from cells that secrete the desired antibody. These antibodies can be screened for binding to normal or defective IL-B60, or they can be screened for agonistic or antagonistic activity, for example, mediated through a receptor. The antibodies can be agonistic or antagonistic, for example, by aesthetically blocking the binding to a receptor. These monoclonal antibodies will usually bind at least one KD of about 1 mM, more usually at least about 300 μM, typically at least about 100 μM, more typically at least about 30 μM, preferably at least about 10 μM. μM, and more preferably at least about 3 μM or more. The antibodies of this invention may also be useful in diagnostic applications. As capture or non-neutralizing antibodies, they can be screened for binding ability to the antigens without inhibiting binding to a receptor. As neutralizing antibodies, they can be useful in competitive binding assays. They will also be useful in the detection or quantification of IL-B60 protein or its receptors. See, for example, Chan (ed. 1987) Immunoloqy: A Practical Guide, Academic Press, Orlando, FL; Price and Newman (eds. 1991) Principies and Practice of Immunoassav. Stockton Press, N.Y. and Ngo (ed. 1988) Nonisotopic Immunoassav. Plenum Press, N.Y. Cross-absorptions or other evaluations will identify antibodies that exhibit various spectra of specificities, eg, shared or unique species specificities. In addition, the antibodies, including antigen binding fragments, of this invention, can be potent antagonists that bind to the antigen and inhibit functional binding, for example, to a receptor that can produce a biological response. They can also be potent as non-neutralizing antibodies, and can be coupled to toxins or radionuclides so that when the antibody binds to the antigen, a cell expressing it, for example, on its surface, dies. In addition, these antibodies can be conjugated to drugs or other therapeutic agents, either directly or indirectly by means of a linker, and can effect the objectification of drugs. The antigen fragments can be linked to other materials, in particular polypeptides, such as polypeptides fused or covalently linked to be used as immunogens. An antigen and its fragments can be fused or covalently linked to a variety of immunogens, such as limpet hemocyanin, bovine serum albumin, tetanus toxoid, etc. See Microbioloqy, Hoeber Medical Division, Harper and Row, 1969; Landsteiner (1962) Specificitv of Seroloqical Reactions. Dover Publications, New York; Williams et al. (1967) Methods in Immunology and Immunochemistry, vol. 1, Academic Press, New York; and Harlow and Lane (1988) Antibodies: A Laboratorv Manual. CSH Press, NY, for descriptions of polyclonal antisera preparation methods. In some cases, it is convenient to prepare monoclonal antibodies from several mammalian hosts, such as mice, rodents, primates, humans, etc. The description of techniques for the preparation of said monoclonal antibodies can be found in, for example, Stites et al. (Eds.) Basic and Clinical Immunology (4th ed.), Lange Medical Publications, Los Altos, CA, and references cited in the. same; Harlow and Lane (1988) Antibodies: A Laboratorv Manual. CSH Press, Goding (1986) Monoclonal Antibodies: Principies and Practice (2nd ed.), Academic Press, New York; and in particular in Kohier and Milstein (1975) in Nature 256: 495-497, which discusses a method of generating monoclonal antibodies. Other suitable techniques involve in vitro exposure of lymphocytes to the antigenic polypeptides, or alternatively, to selection of antibody libraries in phage or similar vectors. See, Huse et al. (1989) "Generation of a Large Combinatorial Library of the Immunoglobulin Repertoire in Phage Lambda", Science 246: 1275-1281; and Ward et al. (1989) Nature 341: 544-546. The polypeptides and antibodies of the present invention can be used with or without modification, including chimeric or humanized antibodies. Frequently, polypeptides and antibodies will be labeled by binding, either covalently or non-covalently, a substance that provides a detectable signal. A wide variety of labels and conjugation techniques is known and extensively reported, both in the scientific literature and in patents. Suitable labels include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent moieties, chemiluminescent portions, magnetic particles, and the like. Patents that teach the use of said labels include US Patent No. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241. In addition, recombinant immunoglobulins can be produced; see Cabilly, U.S. Patent No. 4,816,567; Moore et al., U.S. Patent No. 4,642,334; and Queen and others (1989) Proc. Nat'l Acad. Sci. USA 86: 10029-10033. The antibodies of this invention can also be used for affinity chromatography in the isolation of the protein. Columns can be prepared where antibodies are bound to a solid support. See, for example, Wilchek et al. (1984) Meth. Enzvmol 104: 3-55. Conversely, the protein can be used for depletion or cross-absorbances to prepare selectively specific binding compositions. The antibodies raised against each IL-B60 will also be useful to produce anti-idiotypic antibodies. These will be useful in the detection or diagnosis of various immunological conditions related to the expression of the respective antigens.

SAW. Nucleic acids The peptide sequences described and the related reagents are useful in the detection, isolation or identification of a DNA clone encoding IL-B60, for example, from a natural source. Typically, it will be useful in the isolation of a gene from a mammal, and similar procedures will be applied to isolate genes from other species, e.g., warm-blooded animals, such as birds and mammals. Cross-hybridization will allow isolation of IL-B60 from the same variants, for example polymorphic, or other species. A number of different approaches will be available to successfully isolate a suitable nucleic acid clone. The purified protein or polypeptides are useful for the generation of antibodies by standard methods, as described above. The synthetic peptides or purified protein can be presented to an immune system to generate monoclonal or polyclonal antibodies. See, for example, Coligan (1991) Current Protocols in Immunoloqy and Wiley / Greene; and Harlow and Lane (1989) Antibodies: A Laboratorv Manual, Cold Spring Harbor Press. For example, a specific binding composition could be used for screening an expression library made of a cell line expressing an IL-B60. The screening of intracellular expression can be carried out by various staining or immunofluorescence procedures. The binding compositions could be used to affinity purify or classify cells that express a surface fusion protein. Peptide segments can also be used to predict appropriate oligonucleotides to screen a library. The genetic code can be used to select suitable oligonucleotides useful as probes for screening. See, for example, SEQ ID NO: 1. In combination with polymerase chain reaction (PCR) techniques, synthetic oligonucleotides will be useful in the selection of correct clones from a library. The complementary sequences will also be used as probes, primers, or antisense strands. Several fragments will be particularly useful, for example, coupled with anchored vector or poly-A complementary PCR techniques, or with DNA complementary to other peptides. This invention contemplates the use of isolated DNA or fragments to encode a corresponding biologically active IL-B60 polypeptide, in particular that lacks the portion encoding the untranslated portions of the described sequence. In addition, this invention covers recombinant or isolated DNA encoding a biologically active protein or polypeptide, and which is capable of hybridizing under appropriate conditions to the DNA sequences described herein. Said biologically active protein or polypeptide can be an intact antigen, or fragment, and have an amino acid sequence described in, for example, SEQ ID NO: 2, in particular a mature secretary polypeptide. In addition, this invention covers the use of isolated or recombinant DNA, or fragments thereof, which encode proteins that exhibit high identity to a secreted IL-B60. The isolated DNA may have the respective regulatory sequences on the 5 'and 3' flanks, for example, promoters, enhancers, poly-A addition signals, and others. Alternatively, the expression can be effected by operatively connecting a coding segment to a heterologous promoter, for example, by inserting a promoter upstream of an endogenous gene. An "isolated" nucleic acid is a nucleic acid, for example, an RNA, DNA or a mixed polymer, that is substantially separated from other components that naturally accompany a natural sequence, for example, ribosomes, polymerases and / or flanking genomic sequences of the species of origin. The term encompasses a nucleic acid sequence that has been removed from its natural environment, and includes isolates of cloned or recombinant DNA and chemically synthesized analogs or biologically analogs synthesized by heterologous systems. A substantially pure molecule includes isolated forms of the molecule. Usually, the nucleic acid will be in a vector or fragment less than about 50 kb, usually less than about 30 kb, typically less than about 10 kb, and preferably less than about 6 kb. An isolated nucleic acid will generally be a homogeneous composition of molecules, but in some embodiments, it will contain minor heterogeneity. This heterogeneity is typically found at the polymer ends or non-critical portions for a desired biological function or activity. A "recombinant" nucleic acid is defined either by its production method or by its structure. In reference to its production method, for example, a product made by a process, the method is the use of recombinant nucleic acid techniques, for example, involving human intervention in the nucleotide sequence, typically selection or production. Alternatively, it may be a nucleic acid made by generating a sequence comprising the fusion of two fragments that are not naturally contiguous with each other, but is intended to exclude products of nature, for example, natural mutants. Thus, for example, products made by transforming cells with any non-natural vector are encompassed, as are nucleic acids comprising sequence derived using any synthetic oligonucleotide method. This is often done to replace a codon with a redundant codon encoding the same or a conservative amino acid, while typically entering or removing a sequence recognition site. Alternatively, it is performed to join nucleic acid segments of desired functions, to generate a unique genetic entity comprising a desired combination of functions that are not found in the commonly available natural forms. Restriction enzyme recognition sites are often the target of such artificial manipulations, but other site-specific targets, eg, promoters, DNA replication sites, regulatory sequences, control sequences, or other useful features, may be incorporated by design. A similar concept is proposed for a recombinant, eg, fusion polypeptide. Specifically included are synthetic nucleic acids which, by redundancy of genetic code, encode polypeptides similar to fragments of these antigens, and fusions of sequences of several different species or polymorphic variants. A "fragment" significant in a nucleic acid context is a contiguous segment of at least about 17 nucleotides, generally at least about 22 nucleotides, commonly at least about 29 nucleotides, more often at least about 35 nucleotides, typically at least about 41 nucleotides, usually at least about 47 nucleotides, preferably at least about 55 nucleotides, and in particularly preferred embodiments will be at least about 60 or more nucleotides, eg, 67, 73, 81, 89 , 95, etc. A DNA encoding an IL-B60 protein will be particularly useful for identifying genes, mRNAs and cDNA species that code for similar or related proteins, as well as DNAs that code for homologous proteins of different species. There will be homologs in other species, including primates, rodents, canines, felines and birds. Several IL-B60 proteins should be homologous and are encompassed herein. However, even proteins that have a more distant evolution relationship to the antigen can be isolated without difficulty under appropriate conditions using these sequences, if they are sufficiently homologous. Primate IL-B60 proteins are of particular interest. Recombinant clones derived from genomic sequences, for example, that contain voleurs, will be useful for transgenic studies, including for example, transgenic cells and organisms, and for gene therapy. See, for example, Goodnow (1992) "Transgenic Animáis" in Roitt (ed.) Encvclopedia of Immunoloqy, Academic Press, San Diego, pages 1502 - 1504; Travis (1992) Science 256: 1392-1394; Kuhn et al. (1991) Science 254: 707-710; Capecchi (1989) Science 244, 1288; Robertson (ed. 1987) Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, IRL Press, Oxford; and Rosenberg (1992) J. Clinical Oncology 10: 180-199. Substantial homology, eg, identity, in the context of comparing nucleic acid sequences means either that the segments, or their complementary strands, when compared, are identical when they are optimally aligned, with appropriate insertions or deletions of nucleotides, in at least about 50% of the nucleotides, generally at least about 58%, commonly at least about 65%, often at least about 71%, typically at least about 77%, usually at least about 85%, preferably at least about 95 to 98% or more, and in particular embodiments, as high as about 99% or more of the nucleotides. Alternatively, there is substantial homology when the segments will hybridize under conditions of selective hybridization, to a strand, or its complement, typically using a sequence of IL-B60, for example, in SEQ ID NO: 1. Typically, selective hybridization will occur when there is at least about 55% identity over a stretch of at least about 30 nucleotides, preferably at least about 75% over a stretch of about 25 nucleotides, and more preferably, at least about 90% over about 20 nucleotides. See, Kanehisa (1984) Nuc. Acids Res. 12: 203-213. The identity comparison length, as described, can be over longer stretches, and in certain embodiments will be over a stretch of at least about 17 nucleotides, usually at least about 28 nucleotides. nucleotides, typically at least about 40 nucleotides, and preferably at least about 75 to 100 or more nucleotides. The severe conditions, in reference to homology in the hybridization context, will be severe combined conditions of salt, temperature, organic solvents and other parameters, typically those controlled in hybridization reactions. Severe temperature conditions will usually include temperatures above about 30 ° C, usually above about 37 ° C, typically above about 55 ° C, preferably above about 70 ° C. Severe salt conditions will commonly be less than about 1000 mM, usually less than about 400 mM, typically less than about 250 mM, preferably less than about 150 mM, including about 100, 50, or even 20 mM. However, the combination of parameters is much more important than the measurement of any individual parameter. See, for example, Wetmur and Davidson (1968) J. Mol. Biol. 31: 349-370. Hybridization under severe conditions should provide a background of at least 2 times in depth, preferably at least 3 - 5 or more. For sequence comparison, typically a sequence acts as a reference sequence, to which evaluation sequences are compared. When a sequence comparison algorithm is used, evaluation and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percentage of sequence identity for the evaluation sequence (s), relative to the reference sequence, based on the designated program parameters. The optical alignment of sequences for comparison can be conducted, for example, by the local homology algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2- 482, by the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48-443, by the similarity search method of Pearson and Lipman (1988) Proc Nat'l Acad. Sci. USA 85: 2444, by computerized executions of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package, Gentics Computer Group, 575 Science Dr., Madison, Wl), or by visual inspection (see generally Ausubel and others, above). An example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment of a group of related sequences, using progressive alignments, in pairs, to show relation and percentage of sequence identity. It also diagrams a tree or dendrogram that shows the relationships of groupings used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng and Doolittie (1987) J. Mol. Evol. 35: 351-360. The method used is similar to the method described by Higgins and Sharp (1989) CABIOS 5: 151-153. The program can align up to 300 sequences, each with a maximum length of 5,000 nucleotides or amino acids. The multiple alignment procedure begins with the pairwise alignment of the two most similar sequences, producing a cluster of two aligned sequences. This cluster is then aligned to the next most related sequence or cluster of aligned sequences. Two clusters of sequences are aligned by a simple extension of the alignment of pairs of two individual sequences. The final alignment is achieved by a series of progressive alignments, in pairs.

The program is executed by designating specific sequences and their coordinates of amino acids or nucleotides for regions of sequence comparison, and designating the parameters of the program. For example, a reference sequence can be compared to other evaluation sequences, to determine the percentage relationship of sequence identity using the following parameters: space weight per failure (3.00), space length weight per failure (EYE) and heavy ends spaces. Another example of an algorithm that is suitable for determining the percentage of sequence identity and sequence similarity is the BLAST algorithm, which is described by Altschul et al. (1990) J. Mol. Biol. 215: 403-410. Software to perform BLAST analyzes is available to the public through the National Center for Biotechnology Information (http: www.nebi.nim.nih.gov/). This algorithm involves first identifying pairs of high-ranking sequences (HSPs), identifying short words of length W in the question sequence, which either join or satisfy some positive value threshold T classification when they are aligned with a word of the same value. length in a database sequence. T is referred to as the neighbor word classification threshold (Altschul et al., Above). These initial neighbor word hits act as seeds for the initiation of searches to find longer HSPs that contain them. The word hits are then extended in both directions along each sequence, as long as the cumulative alignment classification can be increased. The extension of word hits in each direction is interrupted when: the classification of cumulative alignment decreases by the amount X of its maximum value achieved; the cumulative classification goes to zero or less, due to the accumulation of one or more residual alignments of negative classification; or the end of each sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLAST program uses as faults a word length (W) of 11, the BLOSUM62 classification matrix alignments (see Henikoff and Henikoff (1989) Proc. Nat'l Acad. Sci. USA 89: 10915) (B) of 50 , expectation (E) of 10, M = 5, N = 4 and a comparison of both strands. In addition to calculating the percentage of sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin and Altschui (1993) Proc. Nat'l Acad. Sci. USA 90: 5873-5787 ). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P (N)), which provides an indication of the probability by which a clash between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the likelihood of a minor sum in a comparison of the assay nucleic acid to the reference nucleic acid is less than about 0J, more preferably less than about 0.01, and with more lower preference of about 0.001.

Another indication that the two polypeptide nucleic acid sequences are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically reactive in cross-over with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, when the two polypeptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under severe conditions, as described below. IL-B60 from other mammalian species can be cloned and isolated by cross-species hybridization of closely related species. Homology may be relatively low among distantly related species, and therefore hybridization of closely related species is advisable. Alternatively, the preparation of an antibody preparation exhibiting less species specificity may be useful in expression cloning approaches VIL Preparation of IL-B60 or complex; mimetics The DNA encoding IL-B60 or fragments thereof can be obtained by chemical synthesis, screening of cDNA libraries, or screening of libraries prepared from a wide variety of cell lines or tissue samples. See, for example, Okayama and Berg (1982) Mol. Cell Biol. 2: 161-170; Gubier and Hoffman (1983) Gene 25: 263-269; and Glover (ed. 1984) DNA Cloninq: A Practical Approach. IRL Press, Oxford. Alternatively, the sequences provided herein provide useful PCR primers, or allow for the synthetic preparation or other preparation of suitable genes encoding an IL-B60; including natural modalities. This DNA can be expressed in a wide variety of host cells for the synthesis of a full-length IL-B60 or fragments which can, in turn, for example, be used to generate monoclonal or polyclonal antibodies; for link studies; for the construction and expression of modified molecules; and for structure / function studies. The vectors, as used herein, comprise plasmids, viruses, bacteriophages, integrable DNA fragments and other vehicles that allow the integration of DNA fragments into the host genome. See, for example, Pouweis et al. (1985 and Supplements) Cloninq Vectors: A Laboratorv Manual, Elsevier, N.Y .; and Rodríguez et al. (eds. 1988) Vectors: A Survev of Molecular Cloninq Vectors and Their Uses, Buttersworth, Boston, MA. For purposes of this invention, the DNA sequences are operatively connected when they are functionally related to each other. For example, DNA for a presequence or secretory leader is operably linked to a polypeptide if it is expressed as a preprotein, or participates in the direction of the polypeptide to the cell membrane, or in the secretion of the polypeptide. A promoter is operably linked to a coding sequence if it controls the transcription of the polypeptide; A ribosomal binding site is operatively connected to a coding sequence if it is located in such a way as to allow translation. Usually, operatively connected means contiguous and in reading frame; however, certain genetic elements such as repressor genes are not contiguously connected, but still bind to operative sequences that in turn control expression. See, for example, Rodríguez et al., Chapter 10, pages 205-236; Balbs and Bolivar (1990) Methods in Enzvmoloqy 185: 14-37; and Ausubel et al. (1993) Current Protocois in Molecular Bioloqv, Greene and Wiley, NY. Representative examples of suitable expression vectors include pCDNAl; pCD, see Okayama et al. (1985) Mol. Cell Biol. 5: 1136-1142; pMCI neo Poly-A, see Thomas et al. (1987) CeJI 51: 503-512; and a baculovirus vector, such as pAC 373 or pAC 610. See, e.g., Miller (1988) Ann. Rev. Microbiol. 42: 177-199. It will often be desired to express an IL-B60 polypeptide in a system that provides a specific or defined glycosylation pattern. See, for example, Luckow and Summers (1988) Bio / Tech noloqy 6: 47-55; and Kaufman (1990) Meth. Enzvmol 185: 487-511. IL-B60, or a fragment thereof, can be designed to be phosphatidyl inositol (P1) bound to a cell membrane, but can be removed from membranes by treatment with a phosphatidyl inositol decomposition enzyme. , for example, phosphatidyl inositol phospholipase-C. This releases the antigen in a biologically active form, and allows the purification by standard procedures of protein chemistry. See, for example, Low (1989) Biochim. Biophvs. Acta 988: 427-454; Tse et al. (1985) Science 230: 1003-1008: v Brunner et al. (1991) J. Cell Biol. 114: 1275-1283. Now that IL-B60 has been characterized, fragments or derivatives thereof can be prepared. by conventional procedures for the synthesis of peptides. These include procedures such as described in Stewart and Young (1984) Solid Phase Peptide Synthesis, Pierce Chemical Co., Rockford, IL.; Bodanszky and Bodanszky (1984) The Practice of Peptide Svnthesis, Springer-Verlag, New York; Bodanszky (1984) The Principies of Peptide Synthesis, Springer-Verlaq, New York; and Villafranca (ed.1991) Techniques in Protein Chemistrv II, Academic Press, San Diego, CA. VIII. USES The present invention provides reagents that will find use in diagnostic applications, as described elsewhere herein, for example, under conditions mediated by IL-B60, or later in the description of diagnostic equipment. The gene may be useful in forensic science, for example, to distinguish rodents from humans, or as a marker to distinguish between different cells that exhibit differential patterns of expression or modification. The compositions provided are reactive useful for, for example, in vitro assays, scientific research and for the synthesis or manufacture of nucleic acids, polypeptides or antibodies. This invention also provides reagents with potential commercial and / or therapeutic significance. 11-B60 (natural or recombinant), fragments thereof and antibodies thereto, together with compounds identified as possessing binding affinity to IL-B60, should be useful as reagents for teaching molecular biology, immunology or physiology. Appropriate equipment can be prepared with the reagents, for example, in practical laboratory exercises in production or use of proteins, antibodies, cloning methods, histology, etc. The reagents will also be useful in the treatment of conditions associated with physiology or abnormal development, including inflammatory conditions. In vitro evaluations may be useful for the detection of the presence or absence of interaction components, which may be correlated with the success of particular treatment strategies. In particular, the modulation of the physiology of several cells, for example, hematopoietic or lymphoid, will be achieved by means of appropriate methods for treatment using the compositions provided herein. See, for example, Thomson (1994; ed.) The Cvtokine Handbook (2nd ed.) Academic Press, San Diego; Metcalf and Nicola (1995) The Hematopoietic Colon Stimulatinq Factors, Cambridge University Press; and Aggarwal and Gutterman (1991) Human Cytokines Blackweil Pub.

For example, a disease or disorder associated with abnormal signaling or abnormal expression by an IL-B60 should be a likely target for an agonist or antagonist. The new cytokine should play a role in the regulation or development of hematopoietic cells, for example, lymphoid cells, which affect immunological responses, for example, inflammation and / or autoimmune disorders. Alternatively, it may affect physiology or vascular development, or neuronal effects. In particular, the cytokine should act as an intermediary, in various contexts, in the synthesis of cytokines by cells, proliferation, etc. Antagonists of IL-B60, such as variants of muteins of a natural form of IL-B60 or blocking antibodies, can provide a selective and powerful way to block immune responses, for example, in situations such as inflammatory or autoimmune responses. See also Samter et al. (Eds.) Immunoloqical Diseases vol. 1 and 2, Littie, Brown and Co. In addition, certain combination compositions would be useful, for example, with other modulators of inflammation. Said other molecules may include steroids, other versions of IL-6 and / or G-CSF, including species variants, or viral homologues, and their respective antagonists. Several abnormal conditions are known in each of the cell types that are shown to produce IL-B60 mRNA by Northern blot analysis. See Berkow (ed.) The Merck Manual of Diacinosis and Therapy, Merck & Co., Rahway, N. J .; Thorn and others Harrison's Principies of Interna! Medicine, McGraw-Hili, N.Y .; and Weatherail et al. (eds.) Oxford Textbook of Medicine. Oxford University Press, Oxford. Many other medical conditions and diseases involve activation by macrophages or monocytes, and many of these will respond to treatment with an agonist or antagonist provided herein. See, for example, Stites and Terr (eds., 1991) Basic and Clinical Immunoloqy Appleton and Lange, Norwalk, Connecticut; and Samter et al. (eds.) Immunoloqical Diseases Littie, Brown and Co. These problems should be amenable to prevention or treatment with the use of the compositions provided herein. The location of pancreatic islets suggests a possible relevance to diabetes. IL-B60, antagonists, antibodies, etc., can be purified and then administered to a patient, veterinarian or human. These reagents can be combined for therapeutic use with additional active or inert ingredients, for example, in conventional pharmaceutically acceptable carriers or diluents, for example, immunogenic adjuvants, together with physiologically harmless stabilizers, excipients or preservatives. These combinations can be sterile filtered and placed in dosage forms, such as by lyophilization in dosing containers, or storage in stabilized aqueous preparations. This invention also contemplates the use of antibodies or binding fragments thereof, including forms that are not complement binders.

Drug screening using IL-B60 or fragments thereof can be performed to identify compounds that have binding affinity, or other relevant biological effects on the functions of IL-B60, including isolation of associated components. Subsequent biological assays can then be used to determine if the compound has intrinsic stimulating activity, and is therefore a blocker or antagonist in that it blocks the activity of the cytokine. Likewise, a compound that has intrinsic stimulating activity can activate the signal path, and therefore is an agonist in that it stimulates the activity of IL-B60. This invention also contemplates the therapeutic use of blocking antibodies for IL-B60 as antagonists, and of stimulant antibodies as agonists. This approach should be particularly useful with other variants of IL-B60 species. The quantities of reagents needed for effective therapy will depend on many different factors, including means of administration, target site, physiological state of the patient and other medications administered. Thus, treatment dosages should be evaluated to optimize safety and efficacy. Typically, dosages used in vitro can provide useful guidance in amounts useful for in situ administration of these reagents. The evaluation in animals of the effective doses for the treatment of particular disorders will provide another indication of prognosis of the human dosage. Various considerations are described, for example, in Gilman et al. (Eds. 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics. 8a. Ed., Pergamon Press; and Reminqtoh's Pharmaceutical Sciences. 17th ed. (1990), Mack Publishing Co., Easton, Penn. Methods for administration are discussed therein and then for oral, intravenous, intraperitoneal or intramuscular administration, transdermal diffusion and others. Pharmaceutically acceptable carriers will include water, saline, pH regulator and other compounds that are described, for example, in the Merck Index, Merck & Co., Rahway, New Jersey. Commonly it would be expected that the dosage ranges are in amounts lower than concentrations of 1 mM, typically lower than concentrations of about 10 μM, usually less than about 100 nM, preferably less than about 10 pM (picomolar), and more preferably less than about 1 fM (femtomolar), with an appropriate carrier. Slow release formulations, or a slow release device, will often be used for continuous or long-term administration. See, for example, Langer (1 990) Science 249: 1527-1533. IL-B60, fragments thereof and antibodies to it or its fragments, antagonists and agonists, can be administered directly to the host to be treated, or in accordance with the size of the compounds, it may be convenient to conjugate them with carrier proteins such as ovalbumin or serum albumin before administration. Therapeutic formulations can be administered in many conventional dosage formulations. While it is possible for the active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation. The formulations typically comprise at least one active ingredient, as defined above, together with one or more acceptable carriers thereof. Each carrier should be both pharmaceutically and physiologically acceptable, in the sense of being compatible with the other ingredients, and not harmful to the patient. The formulations include those suitable for oral, rectal, nasal, topical or parenteral administration (including subcutaneous, intramuscular, intravenous and intradermal). The formulations can conveniently be presented in unit dosage form, and can be prepared by any method well known in the art of pharmacy. See, for example, Gilman et al. (Eds. 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 8a. Ed., Pergamon Press; and Reminqton's Pharmaceutical Sciences, 17th Ed. (1990), Mack Publishing Co., Easton, Penn .; Avis et al. (Eds. 1993) Pharmaceutical Dosaqe Forms: Parenteral Medications. Dekker, New York; Lieberman et al. (Eds. 1990) Pharmaceutical Dosaqe Forms: Tablets. Dekker, New York; and Lieberman et al. (eds. 1990) Pharmaceutical Dosaqe Forms: Disperse Systems. Dekker, New York. The therapy of this invention can be combined, or used, in conjunction with other agents, for example, other cytokines, including IL-6 or G-CSF, or their respective antagonists. Both the natural and recombinant forms of IL-B60 of this invention are particularly useful in kits and assay methods that are capable of screening compounds for detection of protein binding activity. Several methods of automated testing have been developed in recent years, in order to allow the tracking of tens of thousands of compounds in a short period. See, for example, Fodor et al. (1991) Science 251: 767-773, which describes means for evaluating binding affinity by a plurality of defined polymers synthesized on a solid substrate. The development of suitable assays can be greatly facilitated by the ability of large quantities of purified soluble IL-B60 as provided by this invention. Other methods for determining critical residues in receptor interactions of IL-B60-IL-B60 can be used. Mutational analysis can be performed, for example, see Somoza et al. (1993) J. Expti. Med. 178: 549-558, to determine the specific critical residues in the interaction and / or signaling. PHD (Rost and Sander (1994) Proteins 19: 55-72) and DSC (King and Sternberg (1996) Protein Sci. 5: 2298-2310) can provide predictions of secondary structures of a-helix (H), β-strand (E), or spiral (L). The helices A and D are the most important in the interaction of receptors, with the D helix being the most important region. See Table 2. For example, antagonists can normally be found once the antigen has been structurally defined, for example, by tertiary structure information. The evaluation of interaction potentials is now possible with the development of highly automated assay methods, using a purified IL-B60. In particular, new agonists and antagonists will be discovered using the screening techniques described herein. Of particular interest are compounds that demonstrate to have a combined affinity for a spectrum of IL-B60 molecules, for example, compounds that can serve as antagonists for IL-B60 species variants. A method for drug screening utilizes eukaryotic or prokaryotic host cells that are stably transformed with recombinant DNA molecules that express an IL-B60. Cells expressing an IL-B60 isolated from other molecules can be isolated. Said cells, either in a viable or fixed form, can be used for standard link partner binding assays. See, also, Parce et al. (1989) Scienc 246: 243 - 247; and Owicki et al. (1990) Proc. Nat'l Acad. Sci. USA 87: 4007-4011, which describe sensitive methods for detecting cellular responses. Another technique for drug screening involves an approach that provides high throughput screening for compounds that have stable binding affinity for an IL-B60, and is described in detail in Geysen, European Patent Application 84/03564, published on 13 September 1984. First, large amounts of different small peptide evaluation compounds are synthesized on a solid substrate, eg, plastic hairpins or some other suitable surface, see Fodor et al. (1991). Then all the forks are reacted with unpurified, solubilized or purified solubilized IL-B60, and washed. The next step involved detecting bound IL-B60. The rational drug design may also be based on structural studies of the molecular forms of IL-B60 and other effectors or analogues. The effectors may be other proteins that act as intermediates in other functions in response to the linkage, or other proteins that normally interact with IL-B60, for example, a receptor. A means of determining which sites interact with other specific proteins is a determination of physical structure, for example, X-ray crystallography, or 2-dimensional NMR techniques. These will provide guidance as to which amino acid residues form molecular contact regions, as a model, for example, against other models of cytokine receptor. For a detailed description of protein structural determination, see, for example, Blundell and Johnson (1976) Protein Crvstallographv. Academic Press, New York.

IX. Equipment This invention also contemplates the use of IL-B60 proteins, fragments thereof, peptides and their fusion products, in a variety of diagnostic equipment and methods for the detection of the presence of other IL-B60 or link partner. Typically the kit will have a compartment that contains either a defined IL-B60 peptide, or genetic segment or a reagent that recognizes one or the other, eg, fragments of IL-B60 or antibodies. A kit for determining the binding affinity of an evaluation compound to an IL-B60 would typically comprise an evaluation compound; a labeled compound, e.g., a binding partner or antibody having known binding affinity for IL-B60; a source of IL-B60 (natural or recombinant); and a means for separating tagged compound bound from free, such as a solid phase to immobilize the molecule. Once the compounds are screened, those that have adequate binding affinity to the antigen can be evaluated in suitable biological assays, as are well known in the art, to determine whether they act as agonists or antagonists for the signaling pathway of IL-B60 . The capacity of the recombinant IL-B60 peptides also provides well-defined standards for calibrating such assays. A preferred device for determining the concentration of, for example, an IL-B60 in a sample would typically comprise a labeled compound, eg, binding partner or antibody, having known binding affinity for the antigen, a source of cytokine. (natural or recombinant) and a means for separating the free labeling compound from the linkage, for example, a solid phase to immobilize IL-B60. Normally, compartments containing reagents and instructions will be provided.

Antibodies, including antigen binding fragments, specific for IL-B60 or fragments, are useful in diagnostic applications to detect the presence of high levels of IL-B60 and / or its fragments. Such diagnostic assays may employ lysates, living cells, fixed cells, immunofluorescence, cell cultures, body fluids, and may also involve the detection of antigens related to the antigen in serum, or the like. Diagnostic assays can be homogeneous (without a separation step between the free and complex reagent, antigen-binding partner) or heterogeneous (with a separation step). Several commercial assays exist, such as radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA), enzyme immunoassay (EIA), enzyme amplification immunoassay technique (EMIT), fluorescent labeled immunoassay with substrate (SLFIA), and the like . See, for example, Van Vunakis et al. (1980) Meth Enzymol. 70: 1-525; Harlow and Lane (1980) Antibodies: A Laboratorv Manual. CSH Press, NY; and Coligan et al. (eds. 1993) Current Protocois in Immunoloqy, Greeene and Wiley, NY. Anti-idiotypic antibodies may have similar use for the diagnosis of the presence of antibodies against an IL-B60, such as the diagnosis of several abnormal states. For example, overproduction of IL-B60 can result in the production of various immunological reactions that can be diagnostic of abnormal physiological states, in particular in proliferating cell conditions such as cancer or abnormal activation or differentiation. Moreover, the distribution pattern available provides information that the cytokine is expressed in pancreatic islets, suggesting the possibility that the cytokine may be involved in the function of that organ, for example, in a medical condition pertinent to diabetes . Frequently, the reagents for diagnostic tests are supplied as equipment, in order to optimize the sensitivity of the assay. For the present invention, according to the nature of the assay, the protocol and the label, either binding partner or labeled or unlabeled antibody, or labeled IL-B60 is provided. This is usually in conjunction with other additives, such as pH regulator, stabilizers, materials necessary for signal production, such as substrates for enzymes, and the like. Preferably, the equipment will also contain instructions for the proper use and disposal of the contents after use. Typically the equipment has compartments for each useful reagent. Conveniently, the reagents are provided as a dry lyophilized powder, where reagents can be reconstituted in an aqueous medium, providing appropriate concentrations of reagents to effect the assay. Many of the aforementioned constituents of the drug screening and diagnostic assays can be used without modification, or they can be mortified in a variety of ways. For example, labeling can be achieved by covalent or non-covalent attachment of a portion that directly or indirectly provides a detectable signal. In any of these assays, the binding partner, composed of evaluation, IL-B60, or antibodies to them, can be labeled either directly or indirectly. The possibilities for direct labeling include label groups: radiorrhounds such as 125 ?, enzymes, such as peroxidase and alkaline phosphatase, and fluorescent labels (US Patent No. 3,940,475) capable of monitoring the change in fluorescence intensity, change in wavelength , or fluorescence polarization. Possibilities for indirect labeling include biotinylation of a constituent, followed by binding to avidin coupled to one of the preceding label groups. In addition there are numerous methods of separating bound IL-B60 from free, or alternatively, bound free evaluation compound. IL-B60 can be immobilized on several matrices, followed by washing. Suitable matrices include plastic, such as an ELISA plate, filters and beads. See, for example, Coligan et al. (Eds. 1993) Current Protocois in Immunoloqy, Vol. 1, Chapter 2, Greene and Wiley, NY. Other suitable separation techniques include, without limitation, the particle method magnetized fluorescein antibody, described in Rattie et al. (1984) Clin. Chem. 30: 1457-1461, and magnetic particle separation of double antibody, as described in U.S. Patent No. 4,659,678. Methods for the attachment of proteins or their fragments to the various labels have been extensively reported in the literature, and they do not require detailed discussion in the present. Many of the techniques involve the use of activated carboxyl groups, either through the use of carbodiimide or active esters, to form peptide bonds, the formation of thioethers by reaction of a mercapto group with an activated halogen, such as chloroacetyl, or a activated olefin such as maleimide, for binding, or the like. The fusion proteins will also find use in these applications. Another diagnostic aspect of this invention involves the use of oligonucleotide or polynucleotide sequences taken from the sequence of an IL-B60. These sequences can be used as probes for the detection of IL-B60 message levels in samples from patients suspected of having an abnormal condition, for example, inflammatory or autoimmune. Because the cytokine can be a marker or mediator for activation, it may be useful to determine the numbers of activated cells to determine, for example, when additional therapy may be required, for example, in a preventive manner before the effects progress and become significant. The preparation of both DNA and RNA nucleotide sequences, the labeling of the sequences and the preferred size of the sequences have received ample description and discussion in the literature. See, for example, Langer-Safer et al. (1982) Proe. Nat'l. Acad. Sci. 79: 4381 - 4385; Caskey (1987) Science 236: 962-967; and Wilchek and others (1988) Anal. Biochem. 171: 1-32.

Diagnostic equipment that also assess the qualitative or quantitative expression of other molecules are also contemplated. The diagnosis or prognosis may depend on the combination of multiple indications used as markers. Thus, teams can evaluate combinations of markers. See, for example, Viallet et al. (1989) Proqress in Growth Factor Res. 1: 89-97. Other equipment may be used to evaluate other cellular subclasses.

X. Isolation of an IL-B60 receptor Having isolated a ligand from a specific ligand-receptor interaction, there are methods to isolate the receptor. See, Gearing et al. (1989) EMBO J. 8: 3667-3676. For example, means for labeling the cytokine IL-B60 can be determined without interfering with the binding to its receptor. For example, an affinity tag can be fused to either the amino or carboxyl terminus of the ligand. Said label can be a FLAG epitope queue, or, for example, an Ig or Fc domain. An expression library can be screened to determine cytokine specific binding, for example, by cell sorting, or other screening to detect subpopulations that express said binding component. See, for example, Ho et al. (1993) Proc. Nat'l Acad. Sci. USA 90: 11267-11271; and Liu et al. (1994) J. Immunol. 152: 1821-29. Alternatively, a panning method can be used. See, for example, Seed and Aruffo (1987) Proc. Nat'l Acad. Sci. USA 84: 3365-3369.

Protein crosslinking techniques with tag can be applied to isolate cytokine binding partners IL-B60. This would allow the identification of proteins interacting specifically with the cytokine, for example, in a ligand-receptor type manner. Early experiments will be performed to determine whether the known IL-6 or G-CSF receptor components are involved in response (s) to IL-B60. It is also quite possible that these functional receptor complexes can share many or all of the components with a complex of IL-B60 receptors, either a specific receptor subunit, or an accessory receptor subunit. Many modifications and variations of this invention can be made, without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments described herein are offered by way of example only, and the invention should be limited only by the terms of the appended claims, together with the full scope of equivalents to which said claims are entitled.

EXAMPLES I General methods Many of the standard methods below are described, or are referenced, for example, in Maniatis et al. (1982) Molecular Cloninq. A Laboratorv Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY; Sambrook et al. (1989) Molecular Cloning: A Laboratorv Manual (2nd Ed.) Vol. 1 - 3, CSH Press, NY; Ausubel et al., Bioloqv Greene Publishing Associates, Brookiyn, NY; Ausubel et al. (1987 and Supplements) Current Protocols in Molecular Biology Wiley / Greene, NY; Innis et al. (Eds. 1990) PCR Protocols: A Guide to Methods and Applications Academic Press, NY; Bonifacino and others Current Protocois in Cell Bioloqv Wiley, NY; and Doyle et al. Cell and Tissue Culture: Laboratorv Protocols Wiley, NY. Methods for protein purification include methods such as ammonium sulfate precipitation, column chromatography, electrophoresis, centrifugation, crystallization and others. See, for example, Ausubel et al. (1987 and periodic supplements), Deutscher (1990) "Guide to Protein Purification", Methods in Enzvmoloqy vol. 182, and other volumes in this series; Coligan et al. (1995 and supplements) Current Protocois in Protein Science John Wiley and Sons, New York, NY, Matsudaira (ed., 1993) A Practical Guide to Protein and Peptide Purification for Microsequencinq. Academic Press, San Diego, CA; and manufacturer's literature on the use of protein purification products, for example, Pharmacia, Piscataway, NJ, or Bio-Rad, Richmond, CA. The combination with recombinant techniques allows fusion to appropriate segments (epitope tails), for example, to a FLAG sequence or an equivalent, which can be fused, for example, by means of a removable protease sequence. See, for example, Hochuli (1990) "Purification of Recombinant Proteins with Metal Chelate Absorbent" in Setlow (ed.) Genetic Engineering, Principle and Methods 12: 87-98, Plenum Press, NY; and Crowe et al. (1992) QIAexpress: The Hiqh Level Expression & Protein Purification System QUIAGEN, Inc., Chatsworth, CA. Computer sequence analysis is performed, for example, using available software programs, including those from the University of Wisconsin Geneties Computer Group (GCG), Madison, Wi, the NCBI at NIH, and GenBank, NCBI, EMBO and other sources of information. public sequence. Other sources of analysis include, for example, the RASMOL program, see Bazan et al. (1996) Nature. 379: 591; Lodi et al. (1994) Science 263: 1762-1766; Sayle and Miiner-White (1995) TIBS 20: 374-37; and Gronenberg et al. (1991) Protein Engineerinq 4: 263-269; and DSC, see King and Sternberg (1996) Protein Sci. 5: 2298-2310. See, also, Wiikins et al. (eds. 1997) Proteome Research: New Frontiers in Functional Genomics Springer-Verlag, NY; Salzberg et al. (Eds. 1998) Computational Methods in Molecular Bioloqv Elsevier, NY; and Birren et al. (eds. 1997) Genome Analysis: A Laboratory Manual Cold Spring Harbor Press, Cold Spring Harbor, NY. Standard immunological techniques are described, for example, in Hertzenberg et al. (Eds. 1996) Weir's Handbook of Experimental Immunoloqy vols. 1-4, Blackwell Science; Coligan (1991 and updates) Current protocols in Immunloqy Wiley / Greene, NY; and Methods in Enzvmoloqy vol. 70, 73, 74, 84, 92, 93, 108, 116, 121, 132, 150, 162 and 163. Cytokine assays are described, for example, in Thomson (ed., 1994) The Cvtokine Handbook (2nd ed. ) Academic Press, San Diego; Metcalf and Nicola (1995) The Hematopoietic Colonv Stimulating Factors Cambridge University Press; and Aggarwal and Gutterman (1991) Human Cvtokines Blackwell Pub. Assays for vascular biological activities are well known in the art. They will cover angiogenic and angiostatic activities in tumor or other tissues, for example, arterial smooth muscle proliferation (see, for example, Koyoma et al. (1996) Cell 87: 1069-1078), adhesion of monocytes to vascular epithelium (see McEvoy et al. (1997) J. Exp. Med. 185: 2069-2077), etc. See also Ross (1993) Nature 362: 801-809; Rekhter and Gordon (1995) Am. J. Pathol. 147: 668-677; Thyberg et al. (1990) Atherosclerosis 10: 966-9901 and Gumbiner (1996) CeJI 84: 345-357. Assays for the biological activities of neural cells are described, for example, in Wouterlood (ed. 1995) Neuroscience Protocols module 10 , Elsevier; Methods in Neurosciences Academic Press; and Neuromethods Humana Press, Totowa, NJ. The methodology of development systems is described, for example, in Meisami (ed.) Handbook of Human Growth and Developmental Bioloqv CRC Press; and Chríspeels (ed.) Molecular Techniques and Approaches in Developmental Bioloqv Interscience. FACS analyzes are described in Melamed et al. (1990) Flow Cytometry and Sortinq Wiley-Liss, Inc., New York, NY; Shapiro (1988) Practical Flow Cvtometrv Liss, New York, NY; and Robinson et al. (1993) Handbook of Flow Cvtometry Methods Wiley-Liss, New York, NY.

II. Cloning of Human IL-B60 The sequence of the gene is provided in Table 1. The sequence is derived from a human genomic sequence. These sequences allow the preparation of PCR primers, or probes, to determine the cellular distribution of the gene. The sequences allow the isolation of genomic DNA that encodes the message. Using the probe or PCR primers, various tissues or cell types are probed to determine cell distribution. The PCR products are cloned using, for example, a TA cloning kit (Invitrogen). The resulting cDNA plasmids are sequenced from both terms, on an automated sequencer (Applied Biosystems).

III. Cell Expression of IL-B60 Proper probe or primers are prepared for cDNA encoding primate IL-B60. Typically, the probe is labeled, for example, by random primers. The expression is probably in the cell types described, and perhaps also in pancreatic islets. The presence of a leader sequence led to the expectation of finding secreted IL-B60 when expressed in mammalian cells. Transfection of 293T cells with a marked form of hIL-B60 (NL-B60-Etag) did not produce the efficient secretion of IL-B60. Instead, IL-B60 could only be immunoprecipitated from the lysate of the transfected cells. We investigated the possibility that IL-B60 was a compound factor such as IL-12 (p35 / p4O), and thus needed a partner for secretion. Among the non-signaling receptors of the IL-6 family, the recently described, and until now, orphan CLF-1 (NR6) also showed a high level of homology between the human and murine forms (> 95% amino acid identity). ). Based on these observations, a hypothesis was generated that IL-B60 and CLF-1 are partners. Southern Analysis: DNA (5 μg) is digested from a primary amplified cDNA library, with appropriate restriction enzymes to release the inserts, run on a 1% agarose gel and transferred to a nylon membrane (Schleicher and Schuell , Keene, NH). Samples for isolation of human mRNA may include, for example, peripheral blood mononuclear cells (monocytes, T cells, NK cells, granulocytes, B cells), at rest (T100); peripheral blood mononuclear cells, activated with anti-CD3 for 2, 6, 12 h meeting (T101); THO Mot 72 clone, at rest (T102), T cell, THO Mot 72 clone, activated with anti-CD28 and anti-CD3 for 3, 6, 12 h meeting (T103), T cell, THO clone Mot 72, treated anergic with specific peptide for 2, 7, 12 h collected (T104); T cell, clone TH1 HY06, at rest (T107); T cell, clone TH1 HY06, activated with anti-CD28 and anti-CD3 for 3, 6, 12 h meeting (T108); T cell, clone TH1 HY06, anergic treated with specific peptide for 2, 6, 12 h meeting (T109); T cell, clone TH2 HY935, at rest (T110); T cell, clone TH2 HY935, activated with anti-CD28 and anti-CD3 for 2, 7, 12 h meeting (T111); tu tumor cell lines Jurkat and Hut78, at rest (T117); T cell clones, pooled AD130.2, Tc783J2, Tc783.13, Tc783.58, Tc782.69, at rest (T118); clones of T? d cells of chance of T cells, at rest (TU 9); T-cell clone CD28; Splenocytes, at rest (B100); Splenocytes, activated with anti-CD40 and IL-4 (B101); EBV lines of B-cells assembled WT49, RSB, JY, CVIR, 721.221, RM3, HSY, at rest (B102); JY cell line B, activated with PMA and yonomycin for 1, 6 h collected (B103); clones NK 20 collected, at rest (K100); assembled NK 20 clones, activated with PMA and yonomycin for 6 h (K101), NKL clone, peripheral blood derivative of LGL leukemia patient, treated with IL-2 (K106); TF1 hematopoietic precursor line, activated with PMA and yonomycin for 1, 6 h collected (C100); premonocytic line U937, at rest (M100); U937 premonocytic line, activated with PMA and yonomycin for 1, 6 h collected (M101); Elutriated monocytes, activated with LPS, IFN ?, anti-IL-10 during 1, 2, 6, 12, 24 h meeting (M102); elutriated monocytes, activated with LPS, IFN ?, IL-10 during 1, 2, 6, 12, 24 h meeting (M103); elutriated monocytes, activated with LPS, IFN ?, anti-IL-10 for 4, 16 h meeting (M106); elutriated monocytes, activated with LPS, IFN ?, IL-10 for 4, 16 h meeting (M107), elutriated monocytes, activated with LPS for 1 h (M108); elutriated monocytes, activated with LPS for 6 h (M109); DC 70% CD1a +, CD34 +, GM-CSF, TNFa 12 days, at rest (D101); DC 70% CD1a +, CD34 + GM-CSF, TNFa 12 days, activated with PMA and yonomycin for 1 h (D102); DC 70% CD1a +, CD34 + GM-CSF, TNFa 12 days, activated with PMA and yonomycin for 6 h (D103); DC 95% CD1a +, of CD34 + GM-CSF, TNFa 12 days classified FACS, activated with PMA and yonomycin for 1, 6 h collected (D104); DC 95% CD14 +, ex CD34 + GM-CSF, TNFa 12 days classified FACS, activated with PMA and yonomycin for 1, 6 h collected (D105); DC CD1a + CD86 +, of CD34 + GM-CSF, TNFa 12 days classified FACS, activated with PMA and yonomycin for 1, 6 h collected (D106); DC monocytes GM-CSF, IL-4 5 days, at rest (D107); DC of monocytes GM-CSF, IL-4 5 days, at rest (D108); DC of monocytes GM-CSF, IL-4 5 days, activated LPS 4, 16 h collected (D109); DC of monocytes GM-CSF, IL-4 5 days, activated TNFa, supe monocyte for 4, 16 h pooled (D110); epithelial cells, not stimulated; epithelial cells, activated with IL-10; lung fibroblast sarcoma line MRC5, activated with PMA and yonomycin for 1, 6 h collected (C101); Kidney epithelial carcinoma cell line CHA, activated with PMA and yonomycin for 1, 6 h collected (C102). Expression of IL-B60 transcription was very high in elutriated monocytes, activated with LPS, IFN ?, anti-IL-10 for 4, 16 h meeting (M106); elutriated monocytes, activated with LPS, IFNV, anti-IL10 for 1, 2, 6, 12, 24 h meeting (M102); elutriated monocytes, activated LPS for 6 (M109); and elutriated monocytes, activated LPS for 1 h (M108). Samples for expression of mouse mRNA may include, for example, mouse fibroblast L cell line at rest (C200); Transfected cells Braf: ER (Braf fusion to estrogen receptor), control (C201); natural T cells Mel14 + of spleen, at rest (T209); natural Mel14 + spleen cells, stimulated with IFN ?, IL-12 and anti-IL-4, to polarize TH1 cells, exposed to IFN? and IL-4 for 6, 12, 24 h, assembled (T210); natural Mel14 + spleen cells, stimulated with IL-4 and anti-IFN? to polarize Th2 cells, exposed to IL-4 and anti-IFN? for 6, 13, 24 h, assembled (T211); T cells, polarized TH1 (bright Mel14, spleen CD4 + cells, polarized for 7 days with IFN-α and anti IL-4, T200); T cells, polarized TH2 (bright Mel14, spleen CD4 + cells, polarized for 7 days with IL-4 and anti-IFN- ?; T201); T cells, highly TH1 polarized 3x transgenic Balb / C (see Openshaw et al. (1995) J. Exp. Med. 182: 1357-1367, activated with anti-CD3 for 2, 6, 24 h, T202); T cells, highly TH2 polarized 3x transgenic Balb / C (activated with anti-CD3 for 2, 6, 24 h meeting (T203); T cells, highly TH1 polarized 3x C57 bl / 6 transgenic (activated with anti-CD3 during 2, 6, 24 h meeting, T212), highly polarized T2 cells, 3x C57 bi / 6 transgenic (activated with anti-CD3 for 2, 6, 24 h, T213), T cells, highly TH1 polarized (cells Natural CD4 + transgenic Balb / C, polarized 3x with IFN ?, IL-12 and anti-IL-4, stimulated with IGIF, IL-12 and anti-IL-4 for 6, 12, 24 h collected); CD44 - CD25 +, selected from thymus (T204), TH1 D1 J cell clone, resting for 3 weeks after the last stimulation with antigen (T205), TH1 D1 J cell clone, 10 μg / ml stimulated ConA 15 h (T206), TH2 CDC35 T cell clone, resting for 3 weeks after the last antigen challenge (T207), TH2 CDC35 T cell clone, 10 μg / ml ConA stimulated 15 h (T2) 08); line of non-stimulated B cells CH12 (B201); mature non-stimulated B cell leukemia cell line A20 (B200); large B cells not stimulated from spleen (B202); total spleen B cells, activated LPS (B203); dendritic cells enriched with metrizamide, spleen, at rest (D200); bone marrow dendritic cells, at rest (D201); dendritic cells derived from unstimulated bone marrow, depleted with anti-B220, anti-CD3 and anti-Class II, cultured in GM-CSF and IL-4 (D202); dendritic cells derived from bone marrow subjected to depletion with anti-B220, anti-CD3 and anti-Class II, cultured in GM-CSF and IL-4, stimulated with anti-CD40 for 1.5 d, collected (D203); line of monocyte cells RAW 264.7 activated with LPS 4 h (M200); bone marrow macrophages derived with GM and M-CSF (M201); bone marrow macrophages derived with GM-CSF, stimulated with LPS, IFN? and IL-10 for 24 h (M205); bone marrow macrophages derived with GM-CSF, stimulated with LPS, IFN? and anti-IL-10 for 24 h (M206); peritoneal macrophages (M207); macrophage cell line J774, at rest (M202); cell line of macrophages J774 + LPS + anti-IL-10 at 0.5, 1, 3, 6, 12 h meeting (M203); cell line of macrophages J774 + LPS + IL-10 at 0.5, 1, 3, 5, 12 h collected (M204); unstimulated mast cell lines MC-9 and MCP-12 (M208); immortalized endothelial cell line derived from cerebral, non-stimulated microvascular endothelial cells (E200); line of immortalized endothelial cells derived from cerebral microvascular endothelial cells, stimulated overnight with TNFa (E201); line of immortalized endothelial cells derived from cerebral microvascular endothelial cells, stimulated overnight with TNFa (E202); line of immortalized endothelial cells derived from cerebral microvascular endothelial cells, stimulated overnight with TNFa and IL-10 (E203); total aorta of mouse wt C57 bl / 6, total aorta of mouse KO ApoE of 5 months (X207); Total aortic mouse KO ApoE 12 months (X207); wt thymus (0214); total thymus, rag-1 (O208); total kidney, rag-1 (O209); total kidney, mouse NZ B / N; and total heart, rag-1 (O202). Human IL-B60 was found expressed in T cell samples; the ThO Mot72 clone (activated); PBL activated; monocytes; dendritic cells; fetal lung and lung of customary smoker. CLF-1 was found expressed in dendritic cell samples; splenocytes; Th1 cells; fetal lung; and lung. This distribution is consistent with the concept that the complex is important in immune function, for example, dendritic and immune cells, and in the physiology of the lung. Since CLF-1 is necessary for the secretion of IL-B60 in vitro, several libraries of human and mouse cDNAs were screened for determination of co-expression of both mRNA. The highest expression for both was found in adult human splenocytes, T cells, activated monocytes and dendritic cells, and in fetal lung and uterus. In mouse libraries, co-expression was strongest in adult lung.

IV. Chromosome Mapping of IL-B60 An isolated cDNA encoding IL-B60 is used. Chromosome mapping is a standard technique. See, for example, BIOS Laboratories (New Haven, CT) and methods for the use of a hybrid panel of mouse somatic cells with PCR. The human IL-B60 gene has been located in the human chromosome 11.

V. Purification of Protein IL-B60 or Complexes Multiple transfected cell lines are screened for one that expresses the cytokine at a high level, compared to other cells. Alternatively, a recombinant construct can be made with both subunits. Several cell lines are screened and selected for their favorable handling properties. Natural IL-B60 can be isolated from natural sources, or by expression from a transformed cell using an appropriate expression vector. The purification of the expressed protein or complex is achieved by standard procedures, or it can be combined with means designed for effective purification at high efficiency, from lysates or cellular supernatants. FLAG or His6 segments can be used for said purification characteristics. Alternatively, affinity chromatography with specific antibodies can be used; see below. The protein is produced in mammalian expression systems, insect cells, or coli, as desired.

SAW. Isolation of Genes IL-B60 Homologs The cDNA of IL-B60, or another species counterpart sequence, can be used as a hybridization probe to screen a library from a desired source, eg, a primate cell cDNA library . Many different species can be screened both for determining the severity necessary for easy hybridization, and for detecting the presence using a probe. Appropriate hybridization conditions will be used to select clones exhibiting cross-hybridization specificity. Hybridization screening using degenerate probes based on the peptide sequences will also allow the isolation of appropriate clones. Alternatively, the use of appropriate primers for PCR screening will result in enrichment of appropriate nucleic acid clones. Similar methods are applicable to isolate any species variant, allelic or polymorphic. Species variants are isolated using cross-species hybridization techniques based on the isolation of a full-length isolate or fragment of a species such as a probe. Alternatively, antibodies raised against human IL-B60 will be used to screen for cells expressing cross-reactive proteins from an appropriate library, eg, cDNA. The purified protein or defined peptides are useful for generating antibodies by standard methods, as described above. The synthetic peptides or purified protein are presented to an immune system to generate monoclonal or polyclonal antibodies. See, for example, Coligan (1991) Current Protocols in Immunoloqy and Wiley / Greene; and Harlow and Lane (1989) Antibodies: A Laboratory Manual Cold Spring Harbor Press. The resulting antibodies are used for screening, purification or diagnosis, as described.

Vile. Antibodies specific for IL-B60 or complexes The synthetic peptides or purified protein are presented to an immune system to generate monoclonal or polyclonal antibodies. See, for example, Coligan (1991) Current Protocois in Immunology Wiley / Greene; Y Harlow and Lane (1989) Antibodies: A Laboratorv Manual Cold Spring Harbor Press. Polyclonal serum, or hybridomas, can be prepared. In appropriate situations, the present reagent is either labeled, as described above, for example, fluorescence or otherwise, or immobilized to a substrate for panning methods. Immunoselection, immunodepletion and related techniques are available to prepare selective reagents, as desired, for example, for IL-B60 alone, or the complex between the two subunits.

HIV IL-B60 and CLF-1 are coprecipitated A CLF-1 -FLAG construct was prepared in an expression vector. An IL-B6OEtag construct (labeled epitope) was also prepared. Transient transfection in COS cells, either with the IL-B6OEtag construct alone, the CLF-1 -FLAG construct alone, or together. The cells were labeled with 35S methionine. Supernatants and cells were collected. With the co-transfection of cells with soluble IL-B60-Etag and CLF-1 -FLAG receptor, the secretion of both ligand and soluble receptor was greatly improved. Both could be immunoprecipitated with antibodies against either the ligand (anti-Etag) or the receptor (anti-Flag), indicating that IL-B60 and CLF-1 form a soluble cytokine / receptor complex, similar to IL-12 (p35 / p40). ). See Gubler et al. (1991) Proc. Nat'l Acad. Sci. USA 88: 4143-4147; Wolf et al. (1991) J. Immunol. 146: 3074-3081. Consequently, co-expression with a correct partner will produce a dramatic increase in the secretion of the gene products. The co-expression of IL-B60 with other soluble receptors, including Ebi3 (Devergne et al (1996) J. Virol. 70: 1143-1153), IL-12 p40 and sCNTFR (Davis et al. (1991) Science 253: 59 - 63) did not produce an efficient secretion of the ligand. The supernatants were immunoprecipitated either with anti-FLAG M2 (precipitates CLF-1) or with anti-Etag Ab (precipitates 1 L-B60). In IL-BßOEtag transfectants alone, the level of expression in the supernatant detected using the antiEtag antibodies was very low. In contrast, in the double transfectants, the IL-B60Etag and a second labeled band were immunoprecipitated. The second band corresponds to the CLF-1. Therefore, the Etag antibody immunoprecipitates both proteins, for example, forms a complex. In the individual CLF-1 FLAG transfectant, a little CLF-1 FLAG protein is immunoprecipitated with the anti-FLAG M2 Ab. This result is consistent with the other soluble receptors, for example, for p40 component of IL-12. However, in double transfectants, not only more CLF-1 is observed, but now also IL-B60. Immunoprecipitation works in both directions.

IX. IL-B60 binds to CNTFR To identify the signaling receptors for IL-B60 / CLF-1, conditioned medium from 293T cells co-transfected with hlL-B60 and mCLF-1 was added to BA / F3 cells stably transfected with gp130 alone or hgp130 in combination with hIL-6R, hOSMR, hLIFR or hLIFR and hCNTFR, respectively. Only BA / F3 cells expressing gp130, LIFR and CNTFR showed a proliferative response with stimulation with IL-B60 / CLF-1. To analyze the possibility that there is a signaling complex consisting of CNTFR / gp130, or CNTFR / LIFR alone, two soluble fusion proteins were designed that connected either the CNTFR or CLF-1 to IL-B60 via a linker flexible. The so-called similar hypercynins have been shown to be 100-1000 x more active in cells than the cytokine and soluble receptor aggregated separately. See Fischer et al. (1997) Nature Biotechnol .. 15: 142-145. Hyper-CNTFR-IL-B60 was able to induce the proliferation of BAF3 / gp130 / LIFR cells, but not of BAF3 / gp130 cells, showing that the LIFR it is a component of the signaling complex. Stimulation of cells with hyper-CLF-1-IL-B60 did not produce the proliferation of any cell line. This indicated that although it is necessary for the secretion of IL-B60, CLF-1 is not a subunit of the active signaling receptor complex. The involvement of gp130 in the active receptor complex was demonstrated with a neutralizing antibody against gp130, which completely blocked this response. In addition, the analysis of signal transducers in lysates of BA / F3 cells expressing gp130, LIFR and CNTFR, showed that STAT3 is only phosphorylated after stimulation either with co-expressed IL-B60 and CLF-1, or with the CNTFR-IL-B60 fusion protein, but not with the CLF-1-IL-B60 fusion.

X. Evaluation of the Extension of Biological Functions The biological activities of IL-B60 or complex are evaluated on the basis of, for example, the structural and sequence homology between IL-B60 and IL-6 and G-CSF. Initially, the trials that had shown biological activities of IL-6 or G-CSF are examined.

A. Regulation of IL-B60 and CLF-1 after injury to the sciatic nerve. The expression of IL-B60 and CLF-1 in the mouse spinal cord was analyzed in unilateral cross-section of the sciatic nerve, followed by separation of nerve fragments. close and distant, thus avoiding regeneration. At several time points, tissue was collected from the cross-sectional area and analyzed by quantitative PCR to determine the expression of IL-B60 and CLF-1. The cross section of the sciatic nerve produced rapid and lasting up-regulation of ligand and receptor. After 6 hours, IL-B60 and CLF-1 were up-regulated. The expression was still elevated 20 days after the cross section, when compared with nerves not injured or with simulated lesions. In regenerating axons (broken nerves), both IL-B60 and CLF-1 are down-regulated after 12 hours, but while IL-B60 expression almost reaches levels of unharmed nerves after 20 days, the levels of CLF-1 peak after 20 days. This could point to an additional function of CLF-1, possibly in retyping, which begins after two weeks. The cross section of the sciatic nerve in mice lacking GM-CSF and a macrophage response in the nerve, shows that the expression of IL-B60 after 4 days is not altered, compared with normal mice. However, the levels of CLF-1 in those, mice are heterogeneous, ranging from no alteration, to an increase of expression of almost 4 times, compared with normal.

B. Effects on cell proliferation The effect on the proliferation of various cell types is evaluated with various concentrations of cytokine. A dose response analysis is performed in combinations with the related cytokines IL-6, G-CSF, etc. A cytosensing machine can be used, which detects metabolism and cell growth (Molecular Devices, Sunnyvale, CA).

C. Effects on the expression of cell surface molecules in human monocytes Monocytes are purified by negative selection of peripheral blood mononuclear cells from normal healthy donors. Briefly, 3 x 108 mononuclear cells are incubated in ficol bands, on ice with a cocktail of monoclonal antibodies (Becton-Dickinson, Mountain View, CA) consisting, for example, of 200 μl of aCD2 (Leu-5A), 200 μl of aCD3 (Leu-4), 100 μl of aCD8 (Leu 2a), 100 μl of aCD19 (Leu-12), 100 μl of aCD20 (Leu-16), 100 μl of aCD56 (Leu-19), 100 μl of aCD67 (IOM 67; Immunotech, Westbrook, ME) and anti-glycophorin antibody (10F7MN, ATCC Rockville, MD). Antibody-bound cells are washed and then incubated with magnetic beads linked to sheep anti-mouse IgG (Dynal, Oslo, Norway) at a cell count ratio of 20: 1. Antibody-bound cells are separated from monocytes by application of a magnetic field. Subsequently, human monocytes are grown in Yssel's medium (Gemini Bioproducts, Calabasas, CA) containing 1% human AB serum in the absence or presence of IL-B60, IL-6, G-CSF or combinations. The analysis of the expression of cell surface molecules can be carried out by direct immunofluorescence. For example, 2 × 10 5 purified human monocytes are incubated in phosphate buffered saline (PBS) containing 1% human serum on ice for 20 minutes. The cells are pelleted at 200 x g. The cells are again suspended in 20 ml of mAB with FITC or PE label. After an additional 20 minute incubation on ice, the cells are washed in PBS containing 1% human serum, followed by two washes in PBS alone. Cells are fixed in PBS containing 1% paraformaldehyde and analyzed in a FACScan flow cytometer (Becton Dickinson, Mountain View, CA). Exemplary mABs are used, for example: CD11b (anti-mad), CD11c (one gpl 50/95), CD14 (Leu-M3), CD54 (Leu 54), CD80 (anti-BB1 / B7), HLA-DR ( L243) from Becton-Dickinson and CD86 (FUN 1; Pharmingen), CD64 (32.2; Medarex), CD40 (mAb89; Schering-Plow France).

D. Effects of IL-B60 or complex on cytokine production by human monocytes Human monocytes are isolated as described, and cultured in Yssel's medium (Gemini Bioproducts, Calabasas, CA) containing 1% human AB serum in the absence and presence of IL-B60 (material expressed by baculovirus dilution 1/100). In addition, monocytes are stimulated with LPS (E. coli 0127: Difco B8) in the absence or presence of IL-B60, and the concentration of cytokines (IL-1β, IL-6, TNFa, GM-CSF and IL) is determined by ELISA. -10) in the cell culture supernatant.

For intracytoplasmic staining for cytokines, monocytes (1 million / ml) are grown in Yssel's medium, in the absence or presence of IL-B60 and LPS (E. coli 0127: B8 Difco) and 10 mg / ml Brefeldin A (Epicenter technologies Madison Wl) for 12 hours. The cells are washed in PBS and incubated in 2% formaldehyde / PBS solution for 20 minutes at RT. The cells are then washed, resuspended in permeabilization pH buffer (0.5% saponin (Sigma) in PBS / BSA (0.5%) / Azide (1 mM) and incubated for 20 minutes at RT. 2 x 105) are centrifuged and resuspended in 20 ml anti-cytokine conjugated mAbs directly diluted 1: 10 in permeabilization pH regulator for 20 minutes at RT The following antibodies can be used: IL-1a-PE (364 -3B3-14), IL-6-PE (MQ2-13A5), TNFa-PE (Mab11), GM-CSF-PE (BVD2-21C11), and IL-12-PE (C11.5J4; Pharmigen San Diego, CA) Subsequently, the cells are washed twice in permeabilization pH regulator, and once in PBS / BSA / Azide, and analyzed in FACScan flow cytometer (Becton Dickinson-, Mountain View, CA).

E. Effects of IL-B60 on the proliferation of human peripheral blood mononuclear cells (PBMC) Total PBMC is isolated from skin cover of normal healthy donors, by centrifugation through ficol-hypaque, as described (Boyum et al.) . PBMC is cultured in 200 μl Yssel medium (Gemini Bioproducts, Calabasas, CA) containing 1% human AB serum in 96-well plates (Falcon, Becton-Dickinson, NJ), in the absence or presence of IL-B60. The cells are grown in medium alone, or in combination with 100 U / ml IL-2 (R & D Systems) for 120 hours. 3H-Thymidine (0.1 mCi) is added during the last six hours of culture, and the incorporation of 3H-Thymidine by liquid scintillation is determined. Natural, recombinant and fusion proteins would be evaluated for agonist and antagonist activity, in many other biological assay systems, for example, T cells, B cells, NK, macrophages, dendritic cells, hematopoietic progenitors, etc. Due to the structural relationship of IL-6 and G-CSF, trials related to these activities should be analyzed. IL-B60 is evaluated for agonist or antagonist activity on transfected cells expressing G-CSF or IL-6 receptor and controls. See, for example, Ho et al. (1993) Proc. Nat'l Acad. Sci. USA 90, 11267-11271; Ho and others (1995) Mol. Cell Biol. 15: 5043-5053- and Liu et al. (1994) J. Immunol. 152: 1821-1829. IL-B60 is evaluated to determine the effect on macrophage / dendritic cell activation and antigen presentation assays, T-cell cytokine production and proliferation, in response to allogeneic stimulus or antigen. See, for example, Waal Malefyt et al. (1991) J. Exp. Med. 174: 1209-1220; de Waal Malefyt et al. (1991) J. Exp. Med. 174: 915-924; Fiorentino et al. (1991) J. Immunol. 147, 3815-3822; Fiorentino et al. (1991) J. Immunol. 146: 3444-3451; and Groux et al. (1996) J. Exp. Med. 184: 19-29. IL-B60 will also be evaluated for effects on stimulation of NK cells. The tests can be based, for example, on Hsu et al. (1992) Internat. Immunol. 4: 563-569; and Scharwz et al. (1994) J. Immunother. 16: 95-104. Differentiation and growth of B cells will be analyzed, for example, by the methodology described, for example, in Defrance et al. (1992) J. Exp. Med. 175: 671-682; Rousset et al. (1992) Proc. Nat'l. Acad. Sci. USA 89: 1890-1893; including lgG2 and IgA2 change factor assays. Note that, unlike the COS7 supernatants, the supernatants of NIH3T3 and COP apparently do not interfere with human B cell assays.

F. IL-B60 v CLF-1 induces a change in neurotransmitter properties Cholinergic sympathetic neurons innervate at least three different targets: the sweat glands, the vasculature in the skeletal muscle and the periosteum. The mature innervation of sympathetic neurons begins at the end of the first postnatal week, and is characterized by the appearance of cholineric properties. Cultures of sympathetic neurons were analyzed to determine the induction of different neuromodulators, which specify the cholinergic type. Cholecystokinin (CCK), vasoactive intestinal polypeptide (VIP), substance P (SP) and somatostatin (SOM) are up-regulated after stimulation of neurons with conditioned medium of co-transfected cells IL-B60 / CLF-1 or the fusion protein CNTFR-IL-B60. Therefore, the complex exhibits a significant function of developmental biology, and may be effective in inducing certain aspects of neural development.

XI. Generation and Analysis of Genetically Altered Animals Transgenic mice can be generated by standard methods. These animals are useful for determining the effects of overexpression of the gene, in specific tissues, or completely throughout the organism. These animals can provide an interesting insight into the development of the animal, or of particular tissues in various stages. In addition, the effect on various responses to biological effort can be evaluated. See, for example, Hogan et al. (1995) Manipulating the Mouse Embryo: A Laboratorv Manual (2nd ed.) Cold Spring Harbor Laboratory Press. Adenovirus techniques are available for the expression of the gene in various cells and organs. See, for example, Hitt et al. (1997) Adv. Pharmacol. 40: 137-195; and literature from Quantum Biotechnologies, Montreal, Canada. Animals may be useful for determining the effects of the gene on various physiologically functional or developmental animal systems.

The genomic structure for mouse IL-B60 has been determined. A strategy for the production of knockout mice IL-B60 can be developed, and appropriate constructions can be made. All references cited herein are incorporated herein by reference, to the same extent as if each publication or individual patent application was specifically and individually indicated to be incorporated as a reference in its entirety for all purposes. Many modifications and variations of this invention can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments described herein are offered by way of example only, and the invention should be limited only by the terms of the appended claims, together with the full scope of equivalents to which said claims are entitled. ' LIST OF SEQUENCES SEQ ID NO: 1 is a primate IL-B60 natural nucleic acid sequence. SEQ ID NO: 2 is a natural amino acid sequence of primate IL-B60. 5 SEQ ID NO: 3 is a natural nucleic acid sequence of rodent IL-B60. SEQ ID NO: 4 is a natural amino acid sequence of rodent IL-B60. SEC ID NO: 5 is a rodent LIF. SEC ID NO: 6 is a primate LIF. SEQ ID NO: 7 is a primate CT-1. SEQ ID NO: 8 is a rodent CT-1. 10 SEC ID NO: 9 is a primate CNTF. SEC ID NO: 10 is a rodent CNTF. SEQ ID NO: 11 is a primate DNAX IL-40. SEC ID NO: 12 is a primate CLF-1. SEQ ID NO: 13 is a rodent CLF-1. ^ c < 110 > Schering Corporation < 120 > Mammalian cytokines; Related Reagents and Methods < 130 > DX0935K < 140 > < 141 > 0 < 150 > US 09 / 267,901 < 151 > 1 1 - 03 - 1999 < 160 > 13 < 170 > Pateptln Ver.2.0 < 210 > 1 < 211 > 1790 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (162) .. (806) < 220 > < 221 mat_ peptide < 222 > (213) .. (806) < 400 > 1 ccgagcgaaa aaaacctgcg agtgggcctg gcggatgsga ttactaaagc ttcgccggag 60 ccgcggctcg ccctcccact ccgccagcct ccgggagagg agccgcaccc ggccggcccg 120 gccccagccc catggacctc cga atg g cagggg actcgegggg tea gcg tgc ctg 176 Met Leu Ala Leu Cys -15 tgc acg egg falls geg cec cec ccc gcc ctc cea cea geg aae cgc ace __ -. Cvs Tnr Val Leu Trp Kis Leu Prc Wing Val Prc Ala Leu Asr. Are Tr.r -10 -5 -1 ggg gac cea ggg ect ggc ccc ecc aec cag aaa acc eat gac cec acc 272 Gly Asp Pro Gly Pro Gly Pro Ser lie Gln Lys Tr.r Tyr Asp Leu Tnr 5 10 15 20 cgc eac ceg gag falls ca ce cec cgc age eeg gee ggg acc cat ceg aac 320 Arg Tyr Leu Glu His Gln Leu Arg Ser Leu Wing Gly Thr Tyr Leu Asn 25 30 35 tac ctg ggc ccc ect tec aac gag cea gac eec aac ect ccc cgc ceg 368 Tyr Leu Gly Pro Pro Phe Asn Glu Pro Asp Phe Asn Pro Pro Arg Leu 0 45 50 ggg gea gag act ctg ccc agg gcc act gtt gac CCg gag gcg egg cga 416 Gly Wing Glu Thr Leu Pro Arg Wing Thr Val Asp Leu Glu Val Trp Arg 55 60 55 age cec aat gac aaa ctg cgg ceg acc cag aac tac gac gcc eac age 464 Ser Leu Asn Asp Lys Leu Arg Leu Thr Gln Asn Tyr Glu Ala Tyr Ser 70 75 B0 falls ctt ctg tgt tac ttg cgt ggc ccc aac cgt cag gct gcc act gct 512 K s Leu Leu Cys Tyr Leu Arg Gly Leu Asn Arg Gln Wing Wing Thr Wing 85 90 95 100 gag ceg cgc cgc age ceg czz cae eec ege acc age cec cag ggc ctg 560 Glu Leu Arg Arg Ser Leu Ala K s Pne Cys Tnr Ser Leu Gln Gly Leu 105 11"" 115 ceg ggc age aet gcg ggc gtc aeg gea gee ceg ggc tac cea ctg ccc 608 Leu Val Mee Ala Ala Leu Gly Tyr Pro Leu Pro 120 2.25 130 cag ccg ctg ect ggg acc gaa czc ace cgg ace ect ggc ect gcc drops 656 Glp Pro Leu Pro Gly Tnr Glu Pro Thr Trp Thr Pro Gly Pro Wing His 135 140 145 agt gac etc cec cag aag aeg sac gac cec tgg cCg cg aag gag ccg 704 Ser Asp Phe Leu Gln Lys Mee Asp Aep Phe Trp Leu Leu Lys Glu Leu 150 155 160 cao acc egg ctg tgg cgc ecg gcc aag gac tte aac cgg ctc aag aag 752 Glr. Thr Trp Leu Trp Arg Be Wing Lys Asp Phe Asn Arg Leu Lys Lys 165 170 175 180 aag atg cag ect cea gea gct gea gtc acc ctg falls ctg ggg gct cat 800 Lys Met Gln Pro Pro Ala Ala Ala Val Thr Leu Kis Leu Gly His Ala 185 190 195 sgc tte tgactectga cccececcec eecgcecccc ceccaaaccc cgcccccact 656 Gly Phe tegegagage cagccctgea tgccaacacc tgttgagcca ggagacagaa gctgtgagcc 916 tctggccctt tcctggaccg gcegggcgeg egaegcgaec agcccegece cceccccacc 9 ~ é ecccaaaggc ctaccgagce ggggaggagg eacageaggc ccegtccege ccegeeecea 1036 caggaageca Cgctcgaggg agcgtgaage ggeccaggee ggcgcagagg -cgcecacggc 1096 ctgcceacca cecctgcttc cccggccagc gcccacccag ccccccagge ggcacatceg 1156 gagggcaggg gccgaggggc caccaccaca caCgccccec cggggcgaag ccccccsgcc 1216 gccccacecC ccceggaCgg gegeegcccc aaccacecca cceaccccca eacacccaae 1276 aacacggegg ecaggaaaca caaeeceaca caaaaagaga cgagaetaac agegcagggt 1336 tggggtctgc attggaggtg ccctataaac cagaagagaa aatactgaaa gcacaggggc 1396 agggacagac cagaccagac ccaggagtct ccaaagcaca gagtggcaaa caaaacc cga 1456 gctgagcatc aggaccttgc etegaatege cceccagtat tacggegcce ceectctgcc 1516 ccceecccca gggeaeccge gggctgccag gctggggagg gcaaccaeag ccacaccaca 1576 ggaeeecceg aaageeeaca aegcageagc aceccggggc geagggeggc agceccccaa 1636 ggcccegccc cccagcccca cccacecaeg aceceaageg cgccgcacca aeatttacee 1696 geeaeeeaee aeetggagae agaegacaee tacegeagaa ceeceaccce Cgtattaaca 1756 aaeaaaaege cegccccaga aaaa 1790 acaaaaaaaa Tyr Glu Ala Tyr Ser His Leu Leu Cys Tyr Leu Arg Gly Leu Asr. Arg 80 85 90 S5 Gln Ala Ala Thr Ala Glu Leu Arg Arg Ser Leu Ala Kis Phe Cys Thr 100 105 110 Be Leu Gln Gly Leu Leu Gly Be lie Wing Gly Val Met Wing Wing Leu 115 120 125 Gly Tyr Pro Leu Pro Gln Pro Leu Pro Gly Thr slu Pro Thr Trp Thr 130 135 140 Pro Gly Pro Wing His Ser Asp Phe Leu Gln Lys Met Asp Asp Phe Trp 145 150 155 Leu Leu Lys Glu Leu Gln Thr Trp Leu Trp Arg Ser Wing Lys Asp Phe 160 165 170 175 Asn Arg Leu Lys Lys Lys Mee Gln Pro Pro Wing Ala Wing Val Thr Leu 180 185 190 is Leu Gly Wing His Gly Phe 195 < 210 > 2 < 21 1 > 215 < 212 > PRT < 213 > primate < 400 > 2 Leu Wing Cys Leu CC \ vs Tnr Val Leu Trp Kl LS Leu Pro Wing Val Pro 10 '? A Leu Asn Arg Thr Gly Asp Pro Gly Pro Gly Pro Ser He Gln Lys 1i "0 15 Tyr Leu Glu His Gln Leu Arg Ser Leu Ala rhr Tyr Asp Leu Thr Arg 25 30 and Thr Tyr Leu Asn Tyr Leu Glv Pro Pro Phe Asn Glu Pro Asp Phe Glv 35 40 45: sn p ^ 0 * -o Aro Leu Glv Ala Glu Thr Leu Pro Arg Ala Thr Val Asp 50 ~ 55 e0 Leu Glu Val Trp Arg Ser Leu Asn Asp Lys Leu Arg Leu Thr Gln Asn 65 ^ 75 < 210 > 3 < 211 > 648 < 212 > DNA < 213 > primate < 220 > < 221 > CDS < 222 > (1 ) . . (645) < 220 > < 221 > mat_peptide < 222 > (52). . (645). < 400 > 3 r- a c "teg falls cec cce gea gtg cea 4e aeg cea gcc ege cta cgc acg g-g - - ^ pro Ala Val Pro Mee Leu Ala Cys Leu Cys ihr - / to Leu T p ^ 10 c cag aaa 96 gee ctt aat cgc ac gga gat ce ggc ect ggc ccc tec aec cag -a.a.a. "ia .ß ,, üsn Arq Thr Gly Asp Pro Gly Pro Gly Pio Ser He Gln Ly 1s5 144 acc acce gac cec ac cgc tac ceg gag cat ca ctc cgc age tta gct Thr Tyr Asp Leu Thr Arg Tyr Leu Glu His Gln Leu Arg Ser Leu Wing 20 25 30 192 ggg acc eac ceg aac tac ctg ggg ccc ect t aac gag ect gac tte Gly Thr Tyr Leu Asn Tyr Leu Gly Pro Pro Phe Asn Glu Pro Asp Phe 4 400 45 35 aat ect cct cg ggg gea gaa act ceg ccc agg gcc acc gtc aac 240 Asn Pro Pro Arg Leu Gly Wing Glu Thr Leu Pro Arg Wing Thr Val Asn 50 55 60 c ttg gaa gtg tgg cga age ctc aat gac agg ctg cgg ctg acc cag aac 86 Leu Glu Val Trp Arg Ser Leu Asn Asp Arg Leu Arg Leu Thr Gln Asn 65 70 75 tat gag gcg tac agt falls ctc ctg tgt tac tgc ggc etc aac cgt 33S Tyr Glu Ala Tyr Ser His Leu Leu Cys Tyr Leu Arg Gly Leu Asn Arg 80 85 90 95 cag gct gcc ac gct gaa ctc cga cgt age ctg gcc cae tte tgt acc 384 Gln Ala Ala Thr Ala Glu Leu Arg Arg Ser Leu Ala His Phe Cys Thr 100 105 110 A n age ctc cag ggc ctg ctg ggc age att gea ggt gtc .atg gcg acg ctt 432 Ser Leu Gln Gly Leu Leu Gly Ser He Wing Gly Val Met Wing Thr Leu 115 120- 125 ggc tac cea ctg ccc cag ccc ctg cea ggg act gag cea gcc tgg gcc 480 Gly Tyr Pro Leu Pro Gl. Pro Leu Pro Gly Thr Glu Pro Wing Trp Wing 130 135 140 ect ggc ect gcc falls agt gac tte ctc cag aag ac ga gat tte tgg 528 Pro Gly Pro Ala His Ser Asp Phe Leu Gln Lys Met Asp Asp Phe Trp 145 150 155. -ctg ctg aag gag ctg cag acc tgg cta tgg cgt tea gcc aag gac tte 576 Leu Leu Lys Glu Leu Glp Thr Trp Leu Trp Arg Ser Ala Lys Asp Phe 160 165 170 175 aac cgg aa aag aag aag cag ect cea gea gct tea gtc acc ctg 624 Asr. Arg Leu Lys Lys Lys Mee Glr. Pro Pro Ala Ala Ser Val Thr Leu 180 185 190 falls ttg gag gcc cat ggt tte tga 648 Kis Leu Glu Ala His Gly Phe 195 < 210 > 4 < 2 > 215 < 212 > PRT < 213 > primate < 400 > 4 Met Leu Wing Cys Leu Cys Thr Val Leu Trp His Leu Pro Wing Val Pro - \ S -10 -5 Wing Leu Asn Arg Thr Gly Asp Pro Gly Pro Gly Pro Ser He Gln Lys -1 1 5 10 15 Thr Tyr Asp Leu Thr Arg Tyr Leu Glu His Gln Leu Arg Ser Leu Wing 20 25 30 Gly Thr Tyr Leu Asn Tyr Leu Gly Pro Pro Phe Asn Glu Pro Asp Phe 35 40 45 Asn Pro Pro Arg Leu Gly Wing Glu Thr Leu Pro Arg Wing Thr Val Asn 50 55 60 Leu Glu Val Trp Arg Ser Leu Asn Asp Arg Leu Arg Leu Thr Glp Asn 65 70 75 Tyr Glu Wing Tyr Ser His Leu Leu Cys Tyr Leu Arg Gly Leu Asn Arg 80 85 90 95 Gln Ala Ala Thr Ala Glu Leu Arg Arg Ser Leu Ala His Phe Cys Thr 100 105 110 Be Leu Gln Gly Leu Leu Gly Be He Wing Gly Val Met Wing Thr Leu 115 120 125 Gly Tyr Pro Leu Pro Gln Pro Leu Pro Gly Thr Glu Pro Wing Trp Wing 130 135 140 Pro Gly Pro Wing His Ser Asp Phe Leu_ Gln Lys Met Asp Asp Phe Trp '145 150 1S5 Leu Leu Lys Glu Leu Gln Thr Trp Leu Trp Arg Ser Wing Lys Asp Phe 160 165 170 175 Asn Arg Leu Lys Lys Lys Met Gln Pro Pro Wing Wing Ser Val Thr Leu 180 185 190 His Leu Glu Ala His Gly Phe 195 • < 210 > 5 < 211 > 203 < 212 > PRT < 213 > rodent < 400 > 5 Met Lys Val Leu Wing Wing Gly He Val Pro Leu Leu Leu Leu Val Leu 1 5 10 15 His Trp Lys His Gly Wing Gly Pro Pro Leu Pro He Thr Pro Val Asn 20 25 30 Wing Thr Cys Wing He Arg His Pro Cys His Gly Asn Leu Met Asn Gln 35 40 45 He Lys Asn Gln Leu Wing Gln Leu Asn Gly Wing Asn Ala Leu Phe 50 55 60 He Ser Tyr Tyr Thr Wing Gln Gly Glu Pro Phe Pro Asn Asn Val Glu 65 70 75 80 L.vs Leu Cys Ala Pro Asr. Mee Tr.r Asp Phe Prc Ser? .-? E K s Gly Asr. 83 9C 95 Gly Thr Glu Lys Thr Lys Leu Val Glu Leu Tyr Arg Mee Val Wing Tyr 100 105 110 Leu Ser Wing Being Leu Thr Asn He Thr Arg Asp Glr. Lys Val Leu Asr. 115 120 125 Pro Thr Ala Val Ser Leu Glr. Val Lys Leu Asn Wing Thr He Asp Val 130 135 140 Mee Arg Gly Leu Leu Ser Asn Val Leu Cys Arg Leu Cys Asr. Lys Tyr 145 150 155 160 Arg Val Gly His Val Asp Val Pro Pro Val Pro Asp Kis As Asp Lys 1SS 170 175 Glu Ala Phe Glr. Arg Lys Lys Leu Gly Cys Glr. eu Leu Gly Thr Tyr 180 135 190 Lys Glr. Val He Ser Val Val Val Gln Ala Phe 195 200 < 210 > 6 < 211 > 202 < 212 > PRT < 213 > primate < 400 > 6 Met Lys Val Leu Ala Wing Gly Val Val Pro Leu Leu Leu Val Leu His 1"5 10 15 Trp Lys His Gly Wing Gly Pro Pro Leu Pro He Thr Pro Val Asn Wing 20 25 30 Thr Cys Wing He Arg His Pro Cys His Asn Asn Leu Met Asn Gln He 35 40 45 Arg Ser Gln Leu Ala Gln Leu Asn Gly Ser Ala Asn Wing Leu Phe He 50 55 60 Leu Tyr Tyr Thr Wing Gln Gly Glu Pro Phe Pro Asn Asn Leu Asp Lys 65 70 75 80 Leu Cys Gly Pro Asn Val Thr Asp Phe Pro Pro Phe His Wing Asn Gly 85 90 95 Thr Glu Lys Wing Lys Leu Val Glu Leu Tyr Arg He Val Val Tyr Leu 100 105 110 Gly Thr Ser Leu Gly Asn He Thr Arg Asp Gln Lys He Leu Asn Pro 115 120 125 Being Wing Leu Being Leu Being Lys Leu Asn Wing Thr Wing Asp He Leu 130 135 140 Arg Gly Leu Leu Being Asn Val Leu Cys Arg Leu Cys Ser Lys Tyr His 145 150 155 160 Val Gly His Val Asp Val Thr Tyr Gly Pro Asp Thr Ser Gly Lys Asp 165 170 175 Val Phe Gln Lys Lys Lys Leu Gly Cys Gln Leu Leu Gly Lys Tyr Lys 180 185 190 Gln He He Wing Val Leu Wing Gln Ala Phe 195 200 < 210 > 7 < 21 > 201 < 212 > PRT < 213 > primate < 400 > 7 Met Ser Arg Arg Glu Gly Ser Leu Glu Asp Pro Gln Thr Asp Ser Ser 1 5 10 15 Val Ser Leu Leu Pro Kis Leu Glu Ala Lys He Arg Gln Thr Kis Ser 20 25 30 Leu Ala His Leu Leu Thr Lys Tyr Ala Glu Gln Leu Leu Gln Glu Tyr 35 40 45 Val Gln Leu Gln Gly Asp Pro Phe Gly Leu Pro Ser Phe Ser Pro Pro 50 55 60 Arg Leu Pro Val Ala Giy Leu Ser Ala Pro Ala Pro Ser His Ala Gly 65 70 75 80 Leu Pro Val His Glu Arg Leu Arg Leu Asp Wing Wing Wing Leu Wing Wing 85 90 95 Leu Pro Pro Leu Leu Asp Wing Vai Cys Arg Arg Gln Wing Glu Leu Asn 100 105 110 Pro Arg Ala Pro Arg Leu Leu Arg Arg Leu Glu Asp Wing Wing Arg Gln 115 120 125 Wing Arg Wing Leu Gly Wing Wing Val Glu Wing Leu Leu Wing Wing Leu Gly 130 135 140 Wing Wing Asn Arg Gly Pro Arg Wing Glu Pro Pro Wing Wing Thr Wing Be 145 150 155 160 Ala Ala Ser Ala Thr Gly Val Phe Pro Ala Lys Val Leu Gly Leu Arg 165 170 175 Val Cys Gly Leu Tyr Arg Glu Trp Leu Ser Arg Thr Glu Gly Asp Leu 180 185 190 Gly Gin Leu Leu Pro Gly Gly Ser Wing 195 200 < 210 > 8 < 211 > 203 < 212 > PRT < 213 > rodent < 400 > 8 Mee Ser Gln Arg Glu Gly Ser Leu Glu Asp Kis Gin Thr Asp Ser Ser 1 5 10 15 I have been Phe Leu Pro K s Leu Glu Wing Lys He Arg Glr. Thr K s Asn 20 25 30 Leu Wing Arg Leu Leu Thr Lys Tyr Wing Giu Gln Leu Leu Glu Glu Tyr 35 40 45 Vai Gir. Gin Gln Gly Glu Pro Pne Giy Leu Pro Gly Phe Ser Pro Pro 50 55 60 Arg Leu Pro Leu Wing Giy Leu Ser Giy Pro Wing Pro Ser His Wing Gly 65 70 75 80 Leu Pro Val Ser Glu Arg Leu Arg Glr. Asp Ala Ala Ala Leu Ser Val 85 90 95 Leu Pro Ala Leu Leu Asp Ala Vai Arg Arg Arg Gln Ala Glu Leu Asn 100 1C5 110 Pro Arg Ala Pro Arg Leu Leu Are Ser Leu Glu Asp Ala Ala Arg Gln 115 12C 125 Vai A g Ala Leu Gly Ala Ais Vai Glu Tnr Val Leu Ala Ala Leu Gly 130 135 140 Wing Wing Wing Arg Gly Pro Gly Pro Glu Pro Val Thr Val Wing Thr Leu 145 150 155 160 Phe Thr Wing Asn Being Thr Wing Gly He Phe Being Wing Lys Val Leu Gly 165 170 175 Phe Kis Vai Cys Gly Leu Tyr Giy Giu Trp Val Ser Arg Thr Glu Gly 180 135 190 Asp Leu Gly Gln Leu Val Pro Giy Giy Val Wing 195 200 < 210 > 9 < 211 > 200 < 212 > PRT < 213 > primate < 400 > 9 Mee Wing Pne Thr Glu Kis Ser Pro Leu Thr Pro Kis Arg Arg Asp Leu 1 5 10 15 Cys Ser Arg Being He Trp Leu Wing Arg Lys He Arg Being Asp Leu Thr 20 25 30 Wing Leu Thr Glu Being Tyr Vai Lys Kis Gln Gly Leu Asn Lys Asn He 35 40 45 Asn Leu Asp Being Wing Asp Giy Mee Pro Val Wing Being Thr Asp Gin Trp 50 55 50 Ser Glu Leu Thr Glu Wing Giu Arg Leu Gln Glu Asn Leu Gln Wing Tyr 65 70 75 80 Are Thr Phe H s Val Leu Leu Wing Arg Leu Leu Glu Asp Gln Gln Val 85 90 95 Kis Phe Thr Pro Tr.r Giu Gly Asp Pne K s Glr. Ala He His Thr Leu 100 105 110 Leu Leu Glr. Vai Ala Ala Pne Ala Tyr Gin He Giu Glu Leu Met He 115 120 125 Leu Leu Glu Tyr Lys He Pro Arg Asr. Giu Wing Asp Giy Mee Pro He 130 135 140 Asr. Vai Giy Asp Giy Giy Leu Phe Glu Lys Lys Leu Trp Gly Leu Lys 145 150 155 160 Vai Leu Gin Glu Leu Ser Glr. Trp Thr Vai Arg Ser He Kis Asp Leu 165 170 175 Arg Pne He Ser Ser Kis Glr. Thr Gly He Pro Wing Arg Gly Ser His 180 185 190 Tyr He Wing Asn Asn Lys Lys Mee 195 200 < 210 > 10 < 21 1 > 198 < 212 > PRT < 213 > rodent < 400 > 10 Mee Wing Phe Wing Glu Gln Ser Pro Leu Thr Leu Hi = Arg Arg Asp Leu 1 5 ío 15 Cys Ser Arg Ser He Trp Leu Ala Arg Lys He Arg Ser Aso Leu Thr 0 25 30 Ala Leu Met Glu Ser Tyr Val Lys Kis Glr. Gly Leu Asr. Lys Asr. He 35 40 45 Ser Leu Asp Ser Val Asp Gly Val Pro Val Wing Ser T.lr Asp Arg Trp 50 55 60 Ser Glu Mee Thr Glu Wing Glu Arg Leu Gln Glu Asr. Leu Gln Ala Tyr 6S 70 75 80 Are Thr Phe Gln Gly Mee Leu Thr Lys Leu Leu Glu Asp Gln Are Val 85 90 95 His Phe Thr Pro Thr Glu Gly Asp Phe K s GIn Wing He Kis Thr Leu 100 105 110 Tnr Leu Gin Val Wing Wing Pne Wing Tyr Gin Leu Giu Giu Leu Mee Wing 115 120 125 Leu Leu Glu Gln Lys Val Pro Glu Lys Glu Wing Asp Gly Mee Pro Val 130 135 140 Tnr He Gly A.sp Giy Giy Leu Pne Giu Lys Lys Leu Trp Gly Leu Lys 145 150 155 160 Val Leu Glr. Giu Leu Ser Glr. Trp Thr Vai Arg Ser He His Asp Leu 165 170 175 Arg Val He Ser Ser Kis Kis Mee Gly He Ser Wing Kis Glu Ser His 160 185 190 Tyr Gly Ala Lys Gin Mee 195 < 210 > 1 1 < 21 1 > 208 < 212 > PRT < 213 > primate < 400 > 1 1 He z Tnr Kis Leu Ser Leu Leu Z l and Pro Leu Pro Cys Val Arg Thr Ser 10 15 Gir. Glr. Leu Pro Giu Tnr Gin Glr. Val Thr Thr "or Gly Lys Lys Pro 20" 30 Val Ser Val Glyy AArg Arg Glu Val Arg Val Pro Gly Thr Ala Leu Val Pro Ser Leu Leu Ser Vai Ser Val Leu Leu Gln Leu Glp Tw G? - G-, 50 55 60 '"' ' Ser Pro Phe Ser Asp Pro Giy Pne Ser Ala Pro Glu Leu Gln Leu Se == 7C r 7 = 80 Ser Leu Pro Pro Wing Tnr Wing Phe Phe Lys Thr Trp Kis Wing Leu Asp 65 90 95 Asp Giy Giu Arg Leu Ser Leu Ala Glr. Arg Ala He A.sp Pro K s Leu 100 105 110 Gin Leu Vai C-iu Asp A.sp Glr. Be Asp Leu Asr. Pro Giy Ser Pro He 115 120 125 Leu Pro Wing Glp Leu Giy Wing Wing Arg Leu Arg Wing Glr. Gly Pro Leu 130 135 140 Gly Asn Mee Wing Wing He Mee Thr Wing Leu Gly Leu Prc He Pro Pro 145 150 155 160 Giu Glu Asp Thr Pro Giy Leu Wing Wing Phe Gly Wing Be Wing Phe Glu 165 170 175 Arg Lys Cys Arg Gly Tyr Val Val Thr Arg Glu Tyr Gly His Trp Thr 180 1-5 190 Asp Arg Ala Val Arg Asp Leu Ala Leu Leu Lys Ala Lys Tyr Ser Ala 195 200 205 < 210 > 1 1 < 211 > 208 < 212 > PRT < 213 > primate < 400 > 1 1: r Kis Leu Ser Pro Leu Pro Cys Val Arg T -hÍ;: Se: 10 15 Gin Pro Glu Thr Gin Gin Val Thr Thi Pro Gly Lys Pro 20 30 Vai Ser Vai Giy -.rg Are Giu Vai Arg Val Pro Gly Ala Leu Val 40 45 Ser s Leu Ser Vai Ser Val Leu Gir, Leu Gln Tyr Gln Gly 50 60 Ser Prc Ser Aso Pro Gi- • Pne Ser Ala Pro Glu Leu Gln Leu Se- 7C 5 8o Ser Leu Pro Pro Wing Thr Wing Phe Pr.e Lvs Thr Trp Kis Wing Leu Asp S5 90 95 Asp Giy Glu Arg Leu Ser Leu Ala Glr. Arg Ala He Asp Pro Kis Leu ICC 105 110 Gin Leu Vai Giu Asp Asp Glr. Be Asp Leu Asn Pro Giy Ser Pro He 115 120 125 Leu Pro Ala Glr. Leu Giy Wing Wing Arg Leu Arg Wing Gln Gly Pro Leu 130 135 140 Gly Asn Mee Wing Wing He Mee Thr Wing Leu Gly Leu Pro He Pro Pro 145 150 155 160 Glu Glu Asp Thr Pro Gly Leu Wing Wing Phe Gly Wing Be Wing Phe Glu 165 170 175 Arg Lys Cys Ara Gly Tyr Val Val Thr Arg Glu Tyr Gly His Trp Thr 180 1: 5 190 Asp Arg Ala Val Arg Asp Leu Ala Leu Leu Lys Ala Lys Tyr Ser Ala 195 200 205 < 210 > 12 < 211 > 410 < 212 > PRT < 213 > primate < 400 > 12 Me e Pro Al a Gin Se r Al a Arg Arg Prc 10 15 Pro Leu Leu Pro Le1. .e .- Leu Cy? Val Leu Gl and Ala Pro 20 25 30 Are Ala Gly Ser Gly Ala His Tnr Ala Val He Ser Pro Gln Asp Pro 35 40 45 Thr Leu Leu He Gly Ser Ser Leu Leu Ala Thr Cys Ser Val His Gly 50 55 60 Asp Pro Pro Gly Ala Thr Ala Glu Gly Leu Tyr Trp Thr Leu Asn Gly 65 70 75 80 Are Arg Leu Pro Pro Giu Leu Ser Arg Val Leu Asn .Ala Ser Thr Leu 85 90 95 ia Leu Ala Leu Ala Asr. Leu Asr. Gly Ser Arg Glr. Arg Ser Gly Asp 100 105 110 -.sn Leu Val Cys Kis Wing Arg Asp Gly Ser He Leu Wing Gly Ser Cys 115 120 125 > .- ~ 3-- .e._ «:. -_e iCeo- r -. P - Ser Lys Asr. Mee Lys Asp Leu Tnr Cys Are Trp Tnr Pro Giy Wing Kis 145 150 1? 5 160 Giy Giu Tnr Pne Leu Kis Thr Asn Tyr Ser Leu Lys Tyr Lys Leu Are 165 170 1 ~ 5 Trp Tyr Giy Gln Asp Asn Thr Cys Glu Glu Tyr Kis Thr Vai Gly Prc 180 135 190 Kis Ser Cys Kis He Pro Lys Asp Leu Wing Leu Pne Thr Pro Tyr Giu 195 200 205 He Trp Val Glu Wing Thr Asn Are Leu Gly Ser A .ia A.rg Ser Asp Val 210 215 220 Leu Tnr Leu Asp He Leu Asp Val Val Thr Thr Asp Pro Pro Pro Aso 225 230 235 240 Vai Kis Val Ser Arg Val Gly Giy Leu Giu Asp Gln Leu Ser Val Arg 245 250 255 Trp Vai Pro Pro Pro Ala Leu Lys Aso Phe Leu Pne Glr. Ala Lys Tyr 260 265 270 Glr. He Are Tyr Are Vai Giu Asp Ser Val Asp Trp Lys Val Vai Asp 275 2S0 285 Asp Vai Ser Asn Glp Tnr Ser Cys Are Leu Wing Giy Leu Lys Pro Giy 290 295 300 Thr Val Tyr Phe Val Gin Val Are Cys Asn Pro Pne Gly He Tyr Gly 305 310 315 320 Ser Lys Lys Wing Gly He Trp Ser Giu Trp Ser Kis Pro Tnr Ala Wing 325 330 335 Be Tnr Prc Arg Ser Glu Are Prc Gly Pro Gly Giy Gly Wing Cys Glu 340 345 350 Pro Are Giy Gly Glu Pro Ser Gly Pro Val Arg Arlu Glu Leu Lys 355 360 365 Gin Pne Leu Gly Trp Leu Lys His Ala Tyr Cys Ser Asn Leu Ser 370 S ^ S 380 Phe Arg Leu Tyr Asp Gln Trp Are Wing Trp Mee G3n Lys Ser His Lys 385 390 395 400 Thr A.rg Asr. Gin Val Leu Pro Asp Lys Leu 405 410 < 210 > 13 < 21 1 > 407 < 212 > PRT < 213 > rodent < 400 > 13 Arg Pro Leu Ser Ser Leu Trp Ser Pro Leu Leu Leu Cys Val Leu 1 5 10 15 Val Pro Arg Gly Gly Ser Gly Ala K s Thr Ala Val He Ser Pro Glr. 20 25 30 Asp Pro Thr Leu Leu He Gly Be Ser Leu Gin Wing Tnr Cys Ser He 35 40 45 Kis Gly Asp Thr Pro Giy Wing Tnr Wing Giu Gly Leu Tyr Trp Thr Leu 50 55 60 Asr. Giy Arg Arg Leu Pro Ser Leu Ser Arg Leu Leu Asr. Thr Ser Tnr 65 70 p S 80 Leu Ala Leu A.la Leu Ala As Leu Asn Giy Ser Are Glr. Gln Ser Giy e5 90 95 Asp Asn Leu Val Cys Kis Ala Are Asp Gly Ser He Leu Wing Gly Ser 100 105 110 Cys Leu Tyr Val Gly Leu Pro Pro Gi_ Lys Pro As Ne He Ser Cys 115 120 125 Trp Ser Arg Asr. Mee Lys Asp Leu Thr Cys Are Trp Tnr Pro Gly A.la 130 135 140 Kis Giv Giu Thr Phe Leu Kis Thr Asr. Tvr Ser Leu Lys Tvr Lys Leu 145 150 155 160 Are Trp Tyr Gly Gln Asp Asn Thr Cys Giu Glu Tyr His Thr Val Gly 165 170 175 Pro K's Ser Cys Kis He Pro Lys Aso Leu Wing Leu Phe Thr Pro Tyr 130 135 190 Glu He Trp Vai Giu Wing Tnr Asr. Arg Leu Gly Be Wing Arg As Asp 195 200 205 Val Leu Thr Leu Asp Val Leu As Val Val Tnr Thr Asp Pro Pro Pro 210 215 220 -.sp Vai K s Val Ser Arg Val Giy Gly Leu Glu Asp Gln Leu Ser Val : -5 230 235 240 ^ g Trp Val Ser Pro Pro Wing Leu Lys Asp Phe Leu Phe Gln Wing Lys 245 250 255 Tyr Gln He Arg Tyr Arg Val Glu Asp Ser Val Asp Trp Lys Val Val 260 265 270 Asp Asp Vai Ser Asn Gln Thr Ser Cys Arg Leu Wing Gly Leu Lys Pro 275 280 '285 Gly Thr Val Tyr Phe Val Gln Val Arg Cys Asn Pro Phe Gly He Tyr 290 295 300 Gly Ser Lys Lys Wing Gly He Trp Ser Glu Trp Ser Kis Pro Thr Wing 305 310 315 320 Wing Ser Thr Pro Arg Ser Glu Arg Pro Gly Pro Giy Gly Gly Val Cys 325 330 335 Glu Pro Arg Gly Gly Glu Prc Ser Gly Pro Va Arg Arg Glu Leu 340 345 350 Lys Gln Phe Leu Gly Trp Leu Lys Lys His Wing Tyr Cys Ser Asn Leu 355 360 365 Ser Phe Arg Leu Tyr Asp Gln Trp Arg Wing Trp Mee Gin Lys Ser Kis 370 3"> 5 3B0 Lys Thr Arg Asn Gln Asp Giu Gly He Leu Pro Ser Gly Arg Arg Gly 385 390 395 400 Ala Ala Arg Gly Pro A.la Gly 405

Claims (31)

NOVELTY OF THE INVENTION CLAIMS

1. - An isolated soluble complex comprising at least 6 amino acids of the mature protein portion of SEQ ID NO: 2 or 4, and: a) at least 6 amino acids of the mature protein portion of SEQ ID NO: 12 or 13; or b) at least 6 amino acids of the mature protein portion of CNTF-R.

2. The complex of claim 1, wherein said complex: a) comprises a recombinant polypeptide of SEQ ID NO: 2 or 4 mature; or b) comprises a recombinant polypeptide of SEQ ID NO: 12 or 13 mature; c) comprises a recombinant mature CNTF-R polypeptide; or d) comprises both a recombinant polypeptide of SEQ ID NO: 2 or 4 mature, and a recombinant polypeptide of SEQ ID NO: 12 or 13 mature; e) comprises both a recombinant polypeptide of SEQ ID NO: 2 or 4 mature, and a recombinant polypeptide of mature CNTF-R; or f) it is detectably labeled; or g) is in a regulated solution; or h) is in a sterile solution.

3. The complex of claim 1, which: a) comprises a mature IL-B60 polypeptide; b) comprises a mature CLF-1 polypeptide; or c) comprises a mature CNTF-R polypeptide; or d) exhibits at least four non-overlapping segments of at least seven amino acids of SEQ ID NO: 2 or 4; or e) exhibits epitopes of both primate L-B60 and primate CLF-1; or f) exhibits epitopes of both primate L-B60 and primate CNTF-R; g) it is not glycosylated; h) is linked to a solid substrate; or i) is conjugated to another chemical portion; or j) comprises a detection or purification tail, including a FLAG, His6, or Ig sequence.

4.- A team comprising said complex of the claim 1, and: a) a compartment comprising said complex; or b) instructions for the use or disposal of the reagents of said equipment.

5. An isolated or recombinant polypeptide comprising: a) a first segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4, and a second segment comprising at least seven amino acids identical to those of segments of SEQ ID NO: 12 or 13 mature; b) at least two non-overlapping distinctive segments, of at least five amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and a third segment comprising at least seven amino acids identical to the SEC segments ID NO: 12 or 13 mature; c) at least one segment comprising at least seven amino acids identical to the segments of mature SEQ ID NO: 2 6 4, and two non-overlapping distinctive segments, of at least five amino acids identical to the segments of SEQ ID NO: 12 or 13 mature; d) a first segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4, and a second segment comprising at least seven amino acids identical to the mature primate CNTF-R segments; oe) at least two non-overlapping, distinctive segments of at least five amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and a third segment comprising at least seven amino acids identical to the CNTF segments -R mature; of) at least one segment comprising at least seven amino acids identical to the segments of SEQ ID NO: 2 or 4 mature, and two distinctive non-overlapping segments, of at least five amino acids identical to the segments of CNTF- R of mature primate.

6. The polypeptide of claim 5, wherein said distinctive non-overlapping identity segments: a) include one of at least eight amino acids; or b) include one of at least five amino acids and one second of at least six amino acids; or c) include at least three segments of at least four, five and six amino acids, or d) include one of at least twelve amino acids.

7. The polypeptide of claim 5, which: a) comprises a mature IL-B60 sequence; or b) comprises a mature CLF-1 sequence; or c) comprises a mature CNTF-R sequence; or d) exhibits at least four non-overlapping segments, of at least seven amino acids of SEQ ID NO: 2 or 4; or e) has a length of at least about 30 amino acids; f) exhibits epitopes of both primate IL-B60 and primate CLF-1; or g) exhibits epitopes of both primate IL-B60 and primate CNTF-R; or h) is not glycosylated; i) has a molecular weight of at least 30 kD; or j) is a synthetic polypeptide; or k) is linked to a solid substrate; or I) is conjugated to another chemical portion; or m) comprises a detection or purification tail, including a FLAG, His6, or Ig sequence.

8. A composition comprising: a) substantially pure combination of IL-B60 and CLF-1; or b) substantially pure combination of IL-B60 and CNTF-R; c) a sterile polypeptide of claim 5; or d) said polypeptide of claim 5 and a carrier, wherein said carrier is: i) an aqueous compound, including water, saline and / or pH regulator, and / or ii) formulated for oral, rectal, nasal, topical administration or parenteral.

9. A kit comprising a polypeptide of claim 5, and: a) a compartment comprising said polypeptide; or b) instructions for the use or disposal of the reagents of said equipment.

10. A method: a) for making an antiquake recognizing a complex of claim 1, comprising including an immune response in an animal with said complex; or b) for immunoselecting antibodies, comprising contacting a population of antibodies with a complex of claim 1, and separating antibodies that bind, from those that do not bind; or c) for formulating a composition, comprising mixing a complex of claim 1 with a carrier.

11. A binding compound comprising an antigen binding site of an antiquake, the antiquase of which binds specifically to said complex of claim 2d or 2e, but not to any of said mature polypeptides of SEQ ID NO: 2, 4, 12, 13, or CNTF-R.

12. The binding compound of claim 11, wherein: a) said linking compound is: i) in a container; ii) is an Fv, Fab, or Fab2 fragment; or iii) is conjugated to another chemical portion; or b) said antibody; i) is produced against a substantially pure complex of IL-B60 with CLF-1; or ii) is produced against a substantially pure complex of IL-B60 with CNTF-R; iii) is immunoselected; or iv) is a polyclonal antibody; or v) exhibits a Kd to antigen of at least 30 μM; or vi) is attached to a solid substrate, including a bead or plastic membrane; or vii) is in a sterile composition; or viii) is detectably labeled, including a radioactive or fluorescent label.

13. A composition comprising: a) a sterile binding compound of claim 12, or b) said linking compound of claim 12 and a carrier, wherein said carrier is: i) an aqueous compound, including water, saline , and / or pH regulator; and / or ii) formulated for oral, rectal, nasal, topical or parenteral administration.

14. A kit comprising said binding compound of claim 11, and: a) a compartment comprising said binding compound; or b) instructions for the use or disposal of the reagents of said equipment.

15. A method for the production of an antigen: antibody complex, comprising contacting under appropriate conditions, a primate complex comprising: a) polypeptides IL-B60 and CLF-1; or b) IL-B60 and CNTF-R-1 polypeptides with an antibody of claim 11, thus allowing said complex to be formed.

16. The method of claim 15, wherein: a) said complex is purified from other cytokines; or b) said complex is purified from another antibody; or c) said contact is with a sample comprising a cytokine; or d) said contact allows the quantitative detection of said antigen; or e) said contact is with a sample comprising said antibody; or f) said contact allows the quantitative detection of said antibody.

17. An isolated or recombinant nucleic acid: a) encoding said amino acid portions of claim 5; or b) encoding said amino acid portions of claim 5, and comprising a segment of at least 30 contiguous nucleotides of SEQ ID NO: 1 or 3; or c) which will co-express a segment of at least seven contiguous amino acids of SEQ ID NO: 2 or 4, and a segment of at least seven contiguous amino acids of SEQ ID NO: 12 or 13; or d) which will co-express a segment of at least seven contiguous amino acids of SEQ ID NO: 2 or 4, and a segment of at least seven contiguous amino acids of CNTF-R.

18. The nucleic acid of claim 17, which: a) encodes an IL-B60 from a human; or b) encodes a CLF-1 of a human; or c) encodes a human CNTF-R; or d) is an expression vector; or e) also comprises an origin of replication; or f) comprises a detectable label; or g) comprises synthetic nucleotide sequence; or h) is less than 6 kb, preferably less than 3 kb.

19. A cell comprising said recombinant nucleic acid of claim 18.

20. The cell of claim 19, wherein said cell is: a) a prokaryotic cell; or b) a eukaryotic cell; or c) a bacterial cell; or d) a yeast cell; or e) an insect cell; or f) a mammalian cell; or g) a mouse cell; or h) a primate cell; or i) a human cell.

21. A kit comprising said nucleic acid of claim 18, and: a) a compartment comprising said nucleic acid; or b) a compartment further comprising a primate IL-B60 polypeptide; or c) a compartment further comprising a primate CLF-1 polypeptide; or d) a compartment further comprising a primate CNTF-R polypeptide; or e) instructions for the use or disposal of the reagents of said equipment.

22. A method: a) for making a double nucleic acid, comprising contacting a nucleic acid of claim 17 with a complementary nucleic acid, under appropriate conditions, thereby forming said pair; or b) to express a polypeptide, which comprises expressing said nucleic acid of claim 17, thereby producing said polypeptide; or c) to transfect a cell, comprising contacting said cell under appropriate conditions, with said nucleic acid of claim 1, whereby the cell is transfected.

23. An isolated or recombinant nucleic acid encoding at least 5 contiguous amino acids of SEQ ID NO: 12, 13, or primate CNTF-R, and: a) hybridizing under 30 minute wash conditions at 30 ° C and less than 2 M salt, to the coding portion of SEQ ID NO: 1; or b) exhibits identity over a stretch of at least about 30 nucleotides to a primate IL-B60.

24. The isolated nucleic acid of claim 23, wherein: a) said contiguous amino acids are at least 8; or b) said washing conditions are at 45 ° C and / or 500 mM salt; or c) said stretch is at least 55 nucleotides.

25. The recombinant nucleic acid of claim 23, wherein: a) said contiguous amino acids are at least 12; b) said washing conditions are at 55 ° C and / or 150 mM salt; or c) said stretch is at least 75 nucleotides.

26. A method for modulating the physiology or development of tissue culture cells, comprising contacting said cell with an agonist or antagonist of a complex comprising mammalian IL-B60 and: a) CLF-1; or b) CNTF-R.

27. A method for: a) producing a complex of claim 1, comprising co-expressing a recombinant IL-B60 with a recombinant CLF-1 or CNTF-R; or b) increasing the secretion of an IL-B60 polypeptide, which comprises expressing said polypeptide with CLF-1; or c) increasing the secretion of a CLF-1 polypeptide, which comprises expressing said CLF-1 with an IL-B60.

28. The method of claim 27, wherein: a) said < .. 5 increase is at least 3 times; or b) said expression is of an acid • Recombinant nucleic acid V encoding one or both of said polypeptide and CLF-1.

29. A method for tracking for the determination of a receptor that binds said complex of claim 1, comprising 10 contacting said complex with a cell expressing said receptor, under conditions that allow said complex to bind to said receptor, thus forming a detectable interaction.

30. The method of claim 29, wherein said interaction produces a physiological response in said cell. 15

31. The use of an agonist or antagonist of a complex comprising IL-B60 and a) CLF-1 or b) CNTF-R packs the preparation of a pharmaceutical composition to modulate the physiology of the development of a cell.