Pvrig polypeptides and methods of treatment

Abstract

The present invention is directed to PVRIG polypeptides and their uses.

1 . A method of screening for inhibitors of the binding association of PVRIG polypeptide with PVLR2, said method comprising:
 a) providing a surface comprising a first ligand protein comprising one of PVRIG polypeptide or PVLR2 polypeptide;   b) contacting said surface with a candidate agent under physiological conditions, wherein if said candidate agent binds to said first ligand protein it forms a first binding complex,   c) contacting said surface with a second ligand protein comprising the other of PVRIG polypeptide or PVLR2; and   d) determining whether said PVRIG polypeptide and said PVLR2 are bound as an indication of whether said candidate agent inhibits said binding association.   
     
     
         2 . A method of screening for inhibitors of the binding association of PVRIG polypeptide with PVLR2, said method comprising:
 a) providing a cell comprising an exogeneous recombinant nucleic acid encoding a human PVRIG polypeptide, wherein said cell expresses said human PVRIG polypeptide;   b) contacting said cell with a candidate agent and a labeled PVRL2 polypeptide;   c) determining whether said PVRIG polypeptide binds to PVLR2 as an indication of whether said candidate agent inhibits the binding of PVRIG polypeptide with PVLR2.   
     
     
         3 . A method of screening for inhibitors of the binding association of PVRIG polypeptide with PVLR2, said method comprising:
 a) providing a cell comprising an exogeneous recombinant nucleic acid encoding a human PVLR2 polypeptide, wherein said cell expresses said human PVLR2 polypeptide;   b) contacting said cell with a candidate agent and a labeled PVRIG polypeptide;   c) determining whether said PVLR2 polypeptide binds to PVRIG as an indication of whether said candidate agent inhibits the binding of PVRIG polypeptide with PVLR2.   
     
     
         4 . A method of screening for inhibitors of the binding association of PVRIG polypeptide with PVLR2, said method comprising:
 a) providing a test solution comprising:
 i) a PVRIG polypeptide comprising a first FRET label; 
 ii) a PVLR2 polypeptide comprising a second FRET label; 
   c) providing a candidate agent;   d) detecting a FRET signal between said first and second label, wherein a difference in said FRET signal in the presence or absence of said candidate agent indicates that the candidate agent inhibits said binding association.   
     
     
         5 . A method according to any of claims wherein a plurality of candidate agents are tested. 
     
     
         6 . A method according to any of  claims 1  to  5  wherein said candidate agent is a protein. 
     
     
         7 . A method according to  claim 6  wherein said protein is an anti-PVRIG antibody. 
     
     
         8 . A method according to  claim 6  wherein said protein comprises an extracellular domain (ECD) of PVRIG. 
     
     
         9 . A method according to  claim 8  wherein said protein is a fusion protein comprising said ECD and a fusion partner. 
     
     
         10 . A method according to  claim 9  wherein said fusion partner is selected from the group consisting of a human IgG Fc domain and a human serum albumin (HSA). 
     
     
         11 . A method according to any of  claims 1  to  10  wherein said method further comprises:
 a) contacting said candidate agent with a population of cytotoxic T cells (CTLs) under conditions wherein said CTLs would normally be activated; and 
 b) determining the effect of said agent on said activation. 
 
     
     
         12 . A method according to any of  claims 1  to  10  wherein said method further comprises:
 a) contacting said candidate agent with a population of cytotoxic T cells (CTLs); and b) determining the effect of said agent on IFNγ production. 
 
     
     
         13 . A method according to any of  claims 1  to  10  wherein said method further comprises:
 a) contacting said candidate agent with a population of γδ T cells under conditions wherein said γδ T cells would normally be activated; and 
 b) determining the effect of said agent on said activation. 
 
     
     
         14 . A method according to any of  claims 1  to  10  wherein said method further comprises:
 a) contacting said candidate agent with a population of Th1 cells under conditions wherein said Th1 cells would normally be activated; and 
 b) determining the effect of said agent on said activation. 
 
     
     
         15 . A method according to any of  claims 1  to  10  wherein said method further comprises:
 a) contacting said candidate agent with a population of regulatory T cells (Tregs) under conditions and determining the effect of said agent on Treg cell number or activity. 
 
     
     
         16 . A method according to  claims 11  to  15  wherein said determination is done by measuring the presence or absence of increased expression of a protein selected from the group consisting of IFNg, TNFα, GM-CSF, CD25, CD137, CD69, PD1, CD107A, HLA-DR, IL-2, IL-6, IL-4, IL-5, IL-10 and IL-13, wherein increased expression is an indication of activation. 
     
     
         17 . A method of treating a disorder associated with the interaction of PVRIG and PVLR2 comprising administering to a patient a composition comprising a stimulator of PVRIG to effect treatment. 
     
     
         18 . A method according to  claim 17  wherein said treatment is a decrease in immune response. 
     
     
         19 . A method according to  claim 17  wherein said treatment is a decrease in activation of αβ and/or γδ T cells. 
     
     
         20 . A method according to  claim 17  wherein said treatment is a decrease in cytotoxic T cell activity 
     
     
         21 . A method according to  claim 17  wherein said treatment is a decrease in NK and/or NKT cell activity. 
     
     
         22 . A method according to  claim 17  wherein said treatment is a decrease of αβ and/or γδ T-cell activity. 
     
     
         23 . A method according to  claim 17  wherein said treatment is a decrease in pro-inflammatory cytokine secretion 
     
     
         24 . A method according to  claim 17  wherein said treatment is a decrease in IL-2 secretion. 
     
     
         25 . A method according to  claim 17  wherein said treatment is a decrease in interferon-  production. 
     
     
         26 . A method according to  claim 17  wherein said treatment is a decrease in Th1 response. 
     
     
         27 . A method according to  claim 17  wherein said treatment is a increase in Th2 response. 
     
     
         28 . A method according to  claim 17  wherein said treatment is an increase in inhibition of T cell activity. 
     
     
         29 . A method according to  claim 17  wherein said treatment is an increase in inhibition of CTL activity. 
     
     
         30 . A method according to  claim 17  wherein said treatment is an increase in inhibition of NK cell activity. 
     
     
         31 . A method according to  claim 17  wherein said treatment is an increase in αβ and/or γδ T cell exhaustion. 
     
     
         32 . A method according to  claim 17  wherein said treatment is a decrease in αβ and/or γδ T cell response. 
     
     
         33 . A method according to  claim 17  wherein said treatment is a decrease in activity of cytotoxic cells. 
     
     
         34 . A method according to  claim 17  wherein said treatment is a reduction in antigen-specific memory responses. 
     
     
         35 . A method according to  claim 17  wherein said treatment is an inhibition of apoptosis or lysis of cells. 
     
     
         36 . A method according to  claim 17  wherein said treatment is a decrease in cytotoxic or cytostatic effect on cells. 
     
     
         37 . A method according to  claim 17  wherein said treatment is a reduction in direct killing of cells. 
     
     
         38 . A method according to  claim 17  wherein said treatment is a decreases in Th17 activity. 
     
     
         39 . A method according to  claim 17  wherein said treatment is an reduction of complement dependent cytotoxicity and/or antibody dependent cell-mediated cytotoxicity. 
     
     
         40 . A method according to any of  claims 17  to  38  wherein said stimulator is selected from the group consisting of a protein and a nucleic acid. 
     
     
         41 . A method according to  claim 40  wherein said protein comprises an extracellular domain (ECD) of PVRIG. 
     
     
         42 . A method according to  claim 40  wherein said protein is a fusion protein comprising said ECD and a fusion partner. 
     
     
         43 . A method according to  claim 42  wherein said fusion partner is selected from the group consisting of a human IgG Fc domain and a human serum albumin (HSA). 
     
     
         44 . A method according to any of  claims 17  to  43  wherein said patient has an immune disorder. 
     
     
         45 . A method according to 44 wherein said wherein said immune disorder is selected from the group consisting of an autoimmune disease, organ transplant rejection and inflammation. 
     
     
         46 . A method according to  claim 45  wherein said autoimmune disease is selected from the group consisting of rheumatoid arthritis, lupus, Inflammatory bowel disease, psoriasis, multiple sclerosis and diabetes type I. 
     
     
         47 . A composition comprising an isolated PVRIG polypeptide consisting of a PVRIG polypeptide ECD domain having at least 95% identity to the ECD domain of an amino acid sequence selected from the group consisting of the sequences depicted in  FIG.  67   ,  FIG.  91    and  FIGS.  92 A to  92 AT . 
     
     
         48 . A composition according to  claim 47  wherein said isolated PVRIG polypeptide has at least 99% identity to an amino acid sequence selected from the group consisting of the sequences depicted in  FIG.  67   ,  FIG.  91    and  FIGS.  92 A to  92 AT . 
     
     
         49 . A composition according to  claim 47  wherein said isolated PVRIG polypeptide is selected from the group consisting of the sequences depicted in  FIG.  67   ,  FIG.  91    and  FIGS.  92 A to  92 AT . 
     
     
         50 . A composition comprising a PVRIG fusion polypeptide comprising:
 a) an ECD from a PVRIG polypeptide; and   b) a covalently attached fusion partner moiety.   
     
     
         51 . A composition according to  claim 50  wherein said fusion partner moiety is selected from the group consisting of a human IgG Fc domain, a human serum albumin (HSA) and a polyethylene glycol (PEG). 
     
     
         52 . A composition according to  claim 50  wherein said ECD has an amino acid sequence selected from the group consisting of the sequences depicted in  FIG.  67   ,  FIG.  91    and  FIGS.  92 A to  92 AT . 
     
     
         53 . A composition according to  claims 50  to  52  wherein said PVRIG polypeptide and said fusion partner moiety are directly covalently attached. 
     
     
         54 . A composition according to  claim 53  wherein said fusion partner moiety is a polyethylene glycol (PEG) moiety. 
     
     
         55 . A composition according to  claims 50  to  52  wherein said PVRIG polypeptide and said fusion partner moiety are covalently attached using an exogeneous linker. 
     
     
         56 . A composition according to  claim 55  wherein said exogeneous linker is selected from the group consisting of those depicted in  FIG.  93   . 
     
     
         57 . A composition according to  claim 55  or  56  wherein said exogenous linker has the formula (GGGS)n, wherein n is from 1 to 5. 
     
     
         58 . A composition according to any of  claims 50  to  53  and  55  to  57  wherein said fusion partner moiety is a human serum albumin (HSA). 
     
     
         59 . A composition according to any of  claims 50  to  53  and  55  to  57  wherein said fusion partner moiety is an Fc domain. 
     
     
         60 . A composition according to  claim 59  wherein said Fc domain is a human IgG Fc domain. 
     
     
         61 . A composition according to  claim 60  wherein said human IgG Fc domain is selected from the group consisting of the Fc domain of human IgG1, the Fc domain of human IgG2, the Fc domain of human IgG3 and the Fc domain of human IgG4. 
     
     
         62 . A composition according to  claim 59  wherein said Fc domain is a variant human Fc domain from IgG1 or IgG2. 
     
     
         63 . A composition according to any of  claims 50  to  62  further comprising a pharmaceutically acceptable carrier. 
     
     
         64 . A method of suppressing T cell activation of a patient comprising administering a composition according to any of  claims 50  to  63  to said patient such that said patient's immune response is suppressed as a result of treatment. 
     
     
         65 . A method according to  claim 64  wherein said patient has an immune disorder. 
     
     
         66 . A method according to  claim 65  wherein said immune disorder is selected from the group consisting of an autoimmune disease, and organ transplant rejection. 
     
     
         67 . A method according to  claim 66  wherein said autoimmune disease is selected from the group consisting of rheumatoid arthritis, lupus, Inflammatory bowel disease, psoriasis, multiple sclerosis, Diabetes type I.