Mammalian Cytokines; Related Reagents and Methods

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.

1 . A composition comprising:
 a) both:
 i) a substantially pure polypeptide comprising a plurality of distinct segments of at least 7 contiguous amino acid from IL-12 p40; and 
 ii) a substantially pure polypeptide comprising a plurality of distinct segments of at least 7 contiguous amino acids from IL-B30; 
   b) both:
 i) a substantially pure polypeptide comprising at least 11 contiguous amino acids from IL-12 p40; and 
 ii) a substantially pure polypeptide comprising at least 11 contiguous amino acids from IL-B30; 
   c) a substantially pure polypeptide comprising both:
 i) a plurality of distinct segments of at least 7 contiguous amino acids of IL-12 p40; and 
 ii) a plurality of distinct segments of at least 7 contiguous amino acids of IL-B30; or 
   d) a substantially pure polypeptide comprising both:
 i) a segment of at least 11 contiguous amino acids of IL-12 p40; and 
 ii) a segment of at least 11 contiguous amino acids of IL-B30; 
   
     
     
         2 . The composition of  claim 1 :
 a) wherein said plurality of distinct segments of at least 7 contiguous amino acids comprise one segment of at least 9 contiguous amino acids;   b) wherein said plurality of distinct segments of at least 7 contiguous amino acids are both at least 9 contiguous amino acids;   c) wherein said segment of at least 11 contiguous amino acids of IL-12 p40 is at least 15 contiguous amino acids;   d) wherein said segment of at least 11 contiguous amino acids of IL-B30 is at least 15 contiguous amino acids;   e) further comprising a carrier selected from an aqueous compound, including water, saline, and/or buffer;   f) formulated for oral, rectal, nasal, topical, or parenteral administration; or   g) which is sterile composition.   
     
     
         3 . A composition of  claim 1 :
 a) wherein at least one of said polypeptides is:
 i) detectably labeled; 
 ii) recombinantly produced; 
 iii) unglycosylated; 
 iv) denatured; 
 v) attached to a solid substrate; or 
 vi) conjugated to another chemical moiety; 
   b) comprising both:
 i) a substantially pure IL-12 p40 polypeptide; and 
 ii) a substantially pure IL-B30 polypeptide; 
   c) comprising a substantially pure polypeptide comprising IL-12 p40 fused to IL-B30; or   d) combined with IL-18, IL-12, radiation or chemotherapy, an immune adjuvant, or an anti-viral.   
     
     
         4 . A kit comprising a composition of  claim 1 , and:
 a) a compartment comprising said polypeptide; or   b) instructions for use or disposal of reagents in said kit.   
     
     
         5 . An isolated or recombinant nucleic acid encoding:
 a) both:
 i) a substantially pure polypeptide comprising a plurality of distinct segments of at least 7 contiguous amino acid from IL-12 p40; and 
 ii) a substantially pure polypeptide comprising a plurality of distinct segments of at least 7 contiguous amino acids from IL-B30; 
   b) both:
 i) a substantially pure polypeptide comprising at least 11 contiguous amino acids from IL-12 p40; and 
 ii) a substantially pure polypeptide comprising at least 11 contiguous amino acids from IL-B30; 
   c) a substantially pure polypeptide comprising both:
 i) a plurality of distinct segments of at least 7 contiguous amino acids of IL-12 p40; and 
 ii) a plurality of distinct segments of at least 7 contiguous amino acids of IL-B30; or 
   d) a substantially pure polypeptide comprising both:
 i) a segment of at least 11 contiguous amino acids of IL-12 p40; and 
 ii) a segment of at least 11 contiguous amino acids of IL-B30. 
   
     
     
         6 . The nucleic acid of  claim 5 :
 a) wherein said plurality of distinct segments of at least 7 contiguous amino acids comprise one segment of at least 9 contiguous amino acids;   b) wherein said plurality of distinct segments of at least 7 contiguous amino acids are both at least 9 contiguous amino acids;   c) wherein said segment of at least 11 contiguous amino acids of IL-12 p40 is at least 15 contiguous amino acids;   d) wherein said segment of at least 11 contiguous amino acids of IL-B30 is at least 15 contiguous amino acids;   e) wherein said IL-12 p40 is from a primate;   f) wherein said IL-B30 is from a primate;   g) which is an expression vector;   h) which further comprises an origin of replication;   i) which comprises a detectable label;   j) which comprises synthetic nucleotide sequence;   k) which is less than 6 kb, preferably less than 3 kb; or   l) which is from primate.   
     
     
         7 . A cell comprising said recombinant nucleic acid of  claim 6 . 
     
     
         8 . The cell of  claim 7 , wherein said cell is:
 a) a prokaryotic cell;   b) a eukaryotic cell;   c) a bacterial cell;   d) a yeast cell;   e) an insect cell;   f) a mammalian cell;   g) a mouse cell;   h) a primate cell; or   i) a human cell.   
     
     
         9 . A kit comprising said nucleic acid of  claim 6 , and:
 a) a compartment comprising said nucleic acid;   b) a compartment further comprising a primate IL-12 p40 polypeptide; or   c) a compartment further comprising a primate IL-B30 polypeptide; or   d) instructions for use or disposal of reagents in said kit.   
     
     
         10 . A nucleic acid which hybridizes:
 a) under wash conditions of 30 minutes at 50° C. and less than 1M salt to the natural mature coding portion of primate IL-12 p40; and   b) under wash conditions of 30 minutes at 50° C. and less than 1M salt to the natural mature coding portion of primate IL-B30.   
     
     
         11 . The nucleic acid of  claim 10 , wherein:
 a) said wash conditions for IL-12 p40 are at 60° C. and less than 400 mM salt;   b) said wash conditions for IL-B30 are at 60° C. and less than 400 mM salt;   c) said nucleic acid exhibits identity over a stretch of at least 50 nucleotides to sequence encoding primate IL-12 p40; and/or   d) said nucleic acid exhibits identity over a stretch of at least 50 nucleotides to sequence encoding primate IL-B30.   
     
     
         12 . The nucleic acid of  claim 10 , wherein:
 a) said wash conditions for IL-12 p40 are at 65° C. and less than 150 mM salt;   b) said wash conditions for IL-B30 are at 65° C. and less than 150 mM salt;   c) said nucleic acid exhibits identity over a stretch of at least 90 nucleotides to sequence encoding primate IL-12 p40; and/or   d) said nucleic acid exhibits identity over a stretch of at least 90 nucleotides to sequence encoding primate IL-B30.   
     
     
         13 . An antagonist of IL-12 p40/IL-B30 combined with:
 a) a TNFα antagonist;   b) an IL-12 antagonist;   c) IL-10; or   d) steroids.   
     
     
         14 . A binding compound comprising an antigen binding site from an antibody, which specifically binds to a composition of  claim 1 :
 a) comprising a substantially pure polypeptide comprising both:
 i) a substantially pure IL-12 p40 polypeptide; and 
 ii) a substantially pure IL-B30 polypeptide; or 
   b) comprising a substantially pure polypeptide comprising IL-12 p40 fused to IL-B30; but not to either IL-12 p40 or IL-B30 polypeptide.   
     
     
         15 . The binding compound of  claim 14 , wherein:
 a) said binding compound is in a container;   b) said binding compound is an Fv, Fab, or Fab2 fragment;   c) said binding compound is conjugated to another chemical moiety; or   d) said antibody:
 i) is raised against a composition of  claim 1 ; 
 ii) is immunoselected; 
 iii) is a polyclonal antibody; 
 iv) exhibits a Kd to antigen of at least 30 mM; 
 v) is attached to a solid substrate; including a bead or plastic membrane; 
 vi) is in a sterile composition; or 
 vii) is detectably labeled, including a radioactive or fluorescent label. 
   
     
     
         16 . A kit comprising said binding compound of  claim 15 , and:
 a) a compartment comprising said binding compound; or   b) instructions for use or disposal of reagents in said kit.   
     
     
         17 . A method of producing an antigen:antibody complex, comprising contacting, under appropriate conditions, a primate IL-12 p40/IL-B30 composition with an antibody of  claim 14 , thereby allowing said complex to form. 
     
     
         18 . The method of  claim 17 , wherein:
 a) said complex is purified from other cytokines;   b) said complex is purified from other antibody;   c) said contacting is with a sample comprising a cytokine;   d) said contacting allows quantitative detection of said antigen;   e) said contacting is with a sample comprising said antibody; or   f) said contacting allows quantitative detection of said antibody.   
     
     
         19 . A composition comprising:
 a) a sterile binding compound of  claim 14 ; or   b) said binding compound of  claim 14  and a carrier, wherein said carrier is:
 i) an aqueous compound, including water, saline, and/or buffer; and/or 
 ii) formulated for oral, rectal, nasal, topical, or parenteral administration. 
   
     
     
         20 . A method of modulating physiology or development of a cell or tissue comprising contacting said cell with a composition of  claim 1 , or antagonist thereof. 
     
     
         21 . A method of modulating physiology or development of a cell comprising contacting said cell with a composition of  claim 1 , and said contacting results in an increase in production of IFNγ. 
     
     
         22 . The method of  claim 21 , wherein said cell is in a host organism, and said organism exhibits an enhanced Th1 response. 
     
     
         23 . The method of  claim 22 , wherein said Th1 response is selected from:
 a) anti-tumor effect;   b) adjuvant effect;   c) anti-viral effect; or   d) antagonized allergic effect.   
     
     
         24 . A method of modulating physiology or development of a cell in a host organism, comprising administering to said organism a composition of  claim 1 , wherein said contacting results in an:
 a) anti-tumor effect;   b) adjuvant effect;   c) anti-viral effect; or   d) antagonized allergic effect.   
     
     
         25 . The method of  claim 24 , wherein said contacting is in combination with:
 a) IL-18;   b) IL-12;   c) radiation therapy or chemotherapy;   d) an immune adjuvant; or   e) an anti-viral therapeutic.   
     
     
         26 . The method of  claim 24 , wherein said antagonist is an antibody against IL-12 receptor subunit β1. 
     
     
         27 . The method of  claim 24 , wherein said contacting is with an antagonist, and said contacting results in a relative decrease in production of IFNγ. 
     
     
         28 . A method of modulating physiology or development of a cell in a host organism, comprising administering said antagonist to said organism, wherein said contacting results in amelioration of:
 a) an autoimmune condition; or   b) a chronic inflammatory condition.   
     
     
         29 . A method of increasing the secretion of:
 a) a primate IL-B30 comprising expressing said polypeptide with IL-12 p40; or   b) a primate IL-12 p40 comprising expressing said IL-12 p40 with IL-B30.   
     
     
         30 . The method of  claim 28 , wherein:
 a) said increasing is at least 3-fold; or   b) said expressing is of a recombinant nucleic acid encoding IL-B30 and IL-12 p40.   
     
     
         31 . A method of screening for a receptor which binds said composition of  claim 3 , comprising contacting said complex to a cell expressing said receptor under conditions allowing said complex to bind to said receptor, thereby forming a detectable interaction. 
     
     
         32 . The method of  claim 31 , wherein said interaction results in a physiological response in said cell. 
     
     
         33 . A method of modulating the inflammatory response in an animal, said method comprising contacting cells in said animal with a therapeutic amount of:
 a) an agonist of a mammalian IL-B30 protein; or   b) an antagonist of a mammalian IL-B30 protein.   
     
     
         34 . The method of  claim 33 , wherein said:
 a) mammalian IL-B30 protein is a primate protein; or   b) antagonist is an antibody which binds to said mammalian IL-B30;   c) antagonist is an antibody which blocks signaling mediated by mammalian IL-B30.   
     
     
         35 . The method of  claim 33 , wherein said animal exhibits signs or symptoms of an acute phase inflammatory response. 
     
     
         36 . The method of  claim 33 , wherein said sign or symptom is found in skin tissue; lung tissue; gastrointestinal tissue; or liver tissue. 
     
     
         37 . The method of  claim 35 , wherein said sign or symptom is found in skin tissue; lung tissue; gastrointestinal tissue; or liver tissue. 
     
     
         38 . The method of  claim 33 , wherein said modulating is accelerating maturation of neutrophils into platelets. 
     
     
         39 . The method of  claim 33 , wherein said modulating has an effect on IgA. 
     
     
         40 . The method of  claim 33 , wherein said modulating has an effect on IgG. 
     
     
         41 . The method of  claim 38 , wherein said administering is said agonist. 
     
     
         42 . The method of  claim 39 , wherein said administering is said agonist. 
     
     
         43 . The method of  claim 40 , wherein said administering is said agonist. 
     
     
         44 . The method of  claim 41 , wherein
 a) said agonist is mammalian IL-B30 protein; or   b) said animal is experiencing signs or symptoms of an inflammatory condition.   
     
     
         45 . The method of  claim 42 , wherein
 a) said agonist is mammalian IL-B30 protein; or   b) said animal is experiencing signs or symptoms of an inflammatory condition.   
     
     
         46 . The method of  claim 43 , wherein
 a) said agonist is mammalian IL-B30 protein; or   b) said animal is experiencing signs or symptoms of an inflammatory condition.   
     
     
         47 . The method of  claim 44 , wherein said administering is in combination with:
 a) an anti-inflammatory cytokine agonist or antagonist;   b) an analgesic;   c) an anti-inflammatory agent; or   d) a steroid.   
     
     
         48 . The method of  claim 45 , wherein said administering is in combination with:
 a) an anti-inflammatory cytokine agonist or antagonist;   b) an analgesic;   c) an anti-inflammatory agent; or   d) a steroid.   
     
     
         49 . The method of  claim 46 , wherein said administering is in combination with:
 a) an anti-inflammatory cytokine agonist or antagonist;   b) an analgesic;   c) an anti-inflammatory agent; or   d) a steroid.   
     
     
         50 . A method of inducing the proliferation of memory T-cells by administering IL-B30 or an agonist thereof.