Methods and compositions for modulating the activity of the interleukin-35 receptor complex

Abstract

The receptor for Interleukin 35 (IL-35) is provided. The Interleukin 35 Receptor (IL-35R) comprises a heterodimeric complex of the Interluekin12Rβ2 receptor and the gp130 receptor. Various compositions comprising the IL-35R complex, along with polynucleotides encoding the same and kits and methods for the detection of the same the same are provided. Methods of modulating the activity of IL-35R or modulating effector T cell functions are also provided. Such methods employ various IL-35R antagonists and agonists that modulate the activity of the IL-35R complex and, in some embodiments, modulate effector T cell function. Further provided are methods for screening for IL-35R binding agents and for IL-35R modulating agents. Various methods of treatment are further provided.

1 . A soluble Interleukin 35 receptor (IL-35R) complex comprising:
 a) a first polypeptide comprising the extracellular domain of a gp130 polypeptide or a biologically active variant or fragment thereof; and,   b) a second polypeptide comprising the extracellular domain of an IL12Rβ2 polypeptide or a biologically active fragment or variant thereof;   wherein said soluble complex bind IL-35.   
     
     
         2 . The soluble Interleukin 35 receptor (IL-35R) complex of  claim 1 , wherein
 a) said first polypeptide comprises the extracellular domain of SEQ ID NO:3, an active fragment thereof, or a sequence having at least 80% sequence identity to the extracellular domain of SEQ ID NO:3; and,   b) said second polypeptide comprises the extracellular domain of SEQ ID NO:6 or an active fragment thereof or a sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         3 . A polynucleotide comprising a nucleotide sequence encoding a soluble Interleukin 35 receptor (IL-35R) complex comprising:
 a) a first polynucleotide encoding a first polypeptide comprising the extracellular domain a gp130 polypeptide or a biologically active fragment or variant thereof; and,   b) a second polynucleotide encoding a second polypeptide comprising the extracellular domain of an IL12Rβ2 polypeptide or a biologically active variant or fragment thereof.   
     
     
         4 . The polynucleotide of  claim 3 , wherein,
 a) the first polynucleotide encoding the first polypeptide comprises the extracellular domain of SEQ ID NO:3, an active fragment thereof, or a sequence having at least 80% sequence identity to the extracellular domain of SEQ ID NO:3; and,   b) the second polynucleotide encoding the second polypeptide comprises the extracellular domain of SEQ ID NO:6, an active fragment thereof, or a sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         5 . A mixture of polynucleotides encoding a soluble Interleukin 35 receptor (IL-35R) complex comprising:
 a) a first nucleotide sequence encoding a first polypeptide comprising the extracellular domain of a gp130 polypeptide or biologically active variant or fragment thereof; and,   b) a second nucleotide sequence encoding a second polypeptide comprising the extracellular domain of an IL12Rβ2 polypeptide or a biologically active variant or fragment thereof;   wherein said first and said second polypeptide form the soluble IL-35R complex that bind IL-35.   
     
     
         6 . The mixture of polynucleotides of  claim 5 , wherein
 a) the first nucleotide sequence encoding the first polypeptide comprises the extracellular domain of SEQ ID NO:3, an active fragment thereof or a sequence having at least 80% sequence identity to the extracellular domain of SEQ ID NO:3; and,   b) the second nucleotide sequence encoding the second polypeptide comprises the extracellular domain of SEQ ID NO:6, an active fragment thereof, or a sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         7 . A non-human cell comprising the polynucleotide of  claim 3 . 
     
     
         8 . A transgenic animal having stably integrated into its genome the polynucleotide of  claim 3 . 
     
     
         9 . An isolated cell comprising the polynucleotide of  claim 3 . 
     
     
         10 . A pharmaceutical composition comprising the polypeptide of  claim 1 . 
     
     
         11 . An isolated Interleukin 35 receptor (IL-35R) complex comprising
 a) a first polypeptide comprising a gp130 polypeptide or a biologically active variant or fragment thereof; and,   b) a second polypeptide comprising an Il12Rβ2 polypeptide or a biologically active variant or fragment thereof;   wherein said complex has IL-35R activity.   
     
     
         12 . The isolated Interleukin 35 receptor (IL-35R) complex of  claim 11 , wherein
 a) the first polypeptide comprises SEQ ID NO:3 or a sequence having at least 80% sequence identity to the extracellular domain of SEQ ID NO:3; and,   b) the second polypeptide comprises SEQ ID NO:6 or a sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         13 . An antibody that binds substantially only to the IL-35R complex of  claim 12 . 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The antibody of  claim 13 , wherein
 (a) said antibody is a monoclonal antibody; or,   (b) said antibody is bispecific, wherein a first antigen binding domain specifically interacts with said first polypeptide and said second antigen binding domain specifically interacts with said second polypeptide.   
     
     
         17 . (canceled) 
     
     
         18 . An antibody that binds substantially only to the first polypeptide or a biologically active variant or fragment thereof of the IL-35R complex of  claim 11 , wherein
 (a) said antibody behaves as a specific modulating agent for IL-35R and substantially inhibits IL-35 activation of the IL-35R complex; or,   (b) wherein said antibody behaves as a specific modulating agent for IL-35R and substantially inhibits IL-35 activation of the IL-35R complex.   
     
     
         19 . (canceled) 
     
     
         20 . A mixture of a first and a second antibody comprising:
 a) a first antibody having a first chemical moiety, said first antibody binds substantially only to a first polypeptide comprising a gp130 polypeptide or an active variant or fragment thereof; and,   b) a second antibody having a second chemical moiety, said second antibody binds substantially only to a second polypeptide comprising a IL12Rβ2 polypeptide or a biologically active variant or fragment thereat   c) wherein said first and said second chemical moiety allow for the interaction of said first and said second antibody at an IL-35R complex to be detected.   
     
     
         21 . The mixture of the first and the second antibody of  claim 20 , wherein
 a) said first antibody binds substantially only to the first polypeptide comprising SEQ ID NO:3 or a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:3; and,   b) said second antibody bind substantially only to a second polypeptide comprising SEQ ID NO:6 or the sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . A kit for determining the level of expression of a polynucleotide encoding gp130 and a polynucleotide encoding IL12Rβ2 or a biologically active variant or fragment thereof in a sample comprising
 a) a first polynucleotide capable of specifically detecting or specifically amplifying a polynucleotide encoding a gp130 polypeptide or a biologically active variant or fragment thereof; and, 
 h) a second polynucleotide capable of specifically detecting or specifically amplifying a polynucleotide encoding a IL12Rβ2 polypeptide or a biologically active variant or fragment thereof; 
 wherein said encoded polypeptides form a biologically active IL-35R complex. 
 
     
     
         29 . The kit of  claim 28 , wherein
 a) the first polynucleotide is capable of specifically detecting or amplifying a polynucleotide encoding the amino acid sequence of SEQ ID NO:3 or a sequence having at least 80% sequence identity to SEQ ID NO:3; and,   b) the second polynucleotide is capable of specifically detecting or amplifying a polynucleotide encoding the amino acid sequence of SEQ ID NO:6 or a sequence having at least 80% sequence identity to SEQ ID NO:6.   
     
     
         30 . The kit of  claim 28 , wherein said kit comprises
 a) a first and a second primer that share sufficient sequence homology or complementarity to said first polynucleotide to specifically amplify said first polynucleotide; and,   b) a third and a forth primer that share sufficient sequence homology or complementarity to said second polynucleotide to specifically amplify said second polynucleotide.   
     
     
         31 . The kit of  claim 28 , wherein said kit comprises
 a) a first polynucleotide that can specifically detect said first polynucleotide, wherein said first polynucleotide comprises at least one DNA molecule of a sufficient length of contiguous nucleotides identical or complementary to SEQ ID NO:3; and,   b) a second polynucleotide that can specifically detect said second polynucleotide, wherein said second polynucleotide comprises at least one DNA molecule of a sufficient length of contiguous nucleotides identical or complementary to SEQ ID NO:6.   
     
     
         32 . The kit of  claim 28 , wherein said kit comprises
 a) a first polynucleotide that hybridizes under stringent conditions to the sequence of SEQ ID NO:3; and,   b) a second polynucleotide that hybridizes under stringent conditions to the sequence of SEQ ID NO:6.   
     
     
         33 . A kit for determining the presence of Interleukin 35 Receptor (IL-35R) in a sample comprising an antibody of  claim 13 . 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . A method for detecting an IL-35R complex comprising
 a) contacting a sample with a compound which selectively binds to a IL-35R complex; and   b) detecting a complex comprising the IL-35R complex and the compound; and thereby detecting said IL-35R complex.   
     
     
         38 . The method of  claim 37 , wherein the compound which binds to the polypeptide is an antibody. 
     
     
         39 . A method for
 (a) modulating the activity of an Interleukin 35 receptor (IL-35R) complex or increasing an immune response, or increasing the activity of an effector T cell function in a subject, comprising administering to the subject a therapeutically effective amount of a soluble IL-35R complex of  claim 1 ; or,   (b) treating a subject having a cancer or a chronic inflammatory disease, comprising administering to the subject a therapeutically effective amount of a soluble Interleukin 35 receptor (IL-35R) complex of  claim 1 ; or,   (c) increasing the activity of an effector T cell function in a subject, comprising administering to the subject a therapeutically effective amount of a soluble IL-35R complex of  claim 1 .   
     
     
         40 . (canceled) 
     
     
         41 . (canceled) 
     
     
         42 . A method to identify an Interleukin 35 receptor (IL-35R) binding agent comprising the steps of:
 a) contacting the IL-35R complex, or a cell expressing the IL-35R complex with a candidate compound; and   b) determining whether the IL-35R complex binds to the candidate compound.   
     
     
         43 . The method of  claim 42 , wherein the binding of the candidate compound to the IL-35R complex is detected by a method selected from the group consisting of:
 a) detection of binding by direct detecting of candidate compound/IL-35R binding; or   b) detection of binding using a competition binding assay.   
     
     
         44 . (canceled) 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . (canceled) 
     
     
         50 . A method for
 (a) screening for an Interleukin-35R (IL-35R) modulating agent comprising contacting IL-35R with a candidate compound and determining the effect of the test compound on the activity of the IL-3 5R complex to thereby identify a compound which modulates the activity of the IL-35R complex; or,   (b) modulating the activity of an Interleukin 35 Receptor (IL-35R) comprising contacting a cell expressing said IL-35R with an IL-35R specific binding/modulating agent; or,   (c) modulating the activity of an effector T cell function in a subject, comprising administering to the subject a therapeutically effective amount of an Interleukin 35 Receptor (IL-35R) specific binding/modulating agent or,   (d) treating a subject having a cancer or a chronic inflammatory disease, comprising administering to the subject a therapeutically effective amount of an antagonistic Interleukin 35 Receptor (IL-35R) specific binding/modulating agent or,   (e) treating a subject having an autoimmune or inflammatory disorder comprising administering to the subject a therapeutically effective amount of an agonist Interleukin 35 Receptor (IL-35R) specific binding/modulating agent.   
     
     
         51 . (canceled) 
     
     
         52 . (canceled) 
     
     
         53 . (canceled) 
     
     
         54 . (canceled) 
     
     
         55 . (canceled) 
     
     
         56 . (canceled) 
     
     
         57 . (canceled) 
     
     
         58 . (canceled) 
     
     
         59 . (canceled)