Anti-CCR2 antibodies and methods of use therefor

Abstract

The present invention relates to an antibody or functional fragment thereof which binds to a mammalian (e.g., human) CC-chemokine receptor 2 (CCR2) or a portion of the receptor and blocks binding of a ligand to the receptor. The invention further relates to a method of inhibiting the interaction of a cell bearing mammalian CCR2 with a ligand thereof, and to use of the antibodies and fragments in therapeutic, prophylactic and diagnostic methods.

What is claimed is:  
     
         1 . An antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor.  
     
     
         2 . An antibody or antigen-binding fragment according to  claim 1  wherein said antibody or antigen-binding fragment thereof inhibits one or more functions associated with binding of the ligand to the receptor.  
     
     
         3 . An antibody or antigen-binding fragment thereof according to  claim 1  wherein the mammalian CC-chemokine receptor 2 is a human CC-chemokine receptor 2.  
     
     
         4 . An antibody or antigen-binding fragment thereof according to  claim 1  wherein the ligand is a chemokine.  
     
     
         5 . An antibody or antigen-binding fragment thereof according to  claim 4  wherein the chemokine is selected from the group consisting of MCP-1, MCP-2, MCP-3, MCP-4 and combinations thereof.  
     
     
         6 . An antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antibody or fragment is a monoclonal antibody or fragment thereof.  
     
     
         7 . An antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antibody or fragment is a human antibody or fragment thereof.  
     
     
         8 . An antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antigen-binding fragment is selected from the group consisting of an Fv fragment, an Fab fragment, an Fab′ fragment and an F(ab′) 2  fragment.  
     
     
         9 . An antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antibody or fragment is a humanized antibody or fragment thereof.  
     
     
         10 . An antibody or antigen-binding fragment thereof according to  claim 1 , wherein said antibody or fragment is a recombinant antibody or fragment thereof.  
     
     
         11 . A test kit for use in detecting the presence of a mammalian CC-chemokine receptor 2 in a biological sample comprising 
 a) at least one antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor; and    b) one or more ancillary reagents suitable for detecting the presence of a complex between said antibody or antigen-binding fragment thereof and said mammalian CC-chemokine receptor 2 or a portion thereof.    
     
     
         12 . A method of inhibiting the interaction of a cell bearing mammalian CC-chemokine receptor 2 with a ligand thereof, comprising contacting said cell with an effective amount of an antibody or antigen-binding fragment thereof which binds to mammalian CC-chemokine receptor 2 and inhibits binding of said ligand to the receptor.  
     
     
         13 . A method according to  claim 12  wherein the cell is selected from the group consisting of lymphocytes, monocytes, granulocytes, T cells, basophils, and cells comprising a recombinant nucleic acid encoding CCR2 or a portion thereof.  
     
     
         14 . A method according to  claim 13  wherein the T cells are selected from the group consisting of CD8+ cells, CD25+ cells, CD4+ cells and CD45RO+ cells.  
     
     
         15 . A method according to  claim 12  wherein the ligand is a chemokine.  
     
     
         16 . A method according to  claim 15  wherein the chemokine is selected from the group consisting of MCP-1, MCP-2, MCP-3, MCP-4 and combinations thereof.  
     
     
         17 . A method of inhibiting HIV infection of a cell, comprising contacting a cell with an effective amount of a composition comprising an antibody or antigen-binding fragment thereof which binds to mammalian CC-chemokine receptor 2 and inhibits HIV entry into said cell.  
     
     
         18 . A method according to  claim 17  wherein the cell is selected from the group consisting of lymphocytes, monocytes, macrophages, granulocytes, T cells, and cells comprising a recombinant nucleic acid encoding CCR2 or a portion thereof.  
     
     
         19 . A method according to  claim 18  wherein the T cells are selected from the group consisting of CD8+cells, CD25+cells, CD4+cells and CD45RO+cells.  
     
     
         20 . A method of treating HIV in a patient comprising administering to the patient a composition comprising an effective amount of an antibody or antigen-binding fragment thereof which binds to mammalian CC-chemokine receptor 2 and inhibits HIV entry into said cell.  
     
     
         21 . A method of detecting expression of mammalian CC-chemokine receptor 2 or portion thereof by a cell, comprising: 
 a) contacting a composition comprising a cell to be tested with an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor; and    b) detecting binding of said antibody or antigen-binding fragment thereof,    wherein the binding of said antibody or antigen-binding fragment thereof indicates the presence of said receptor or portion of said receptor on said cell.    
     
     
         22 . The method of  claim 21  wherein the composition is a sample comprising human cells.  
     
     
         23 . A method of detecting a mammalian CC-chemokine receptor 2 or portion of said receptor, comprising: 
 a) contacting a sample to be tested with an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor under conditions appropriate for binding of said antibody or fragment thereof thereto; and    b) detecting or measuring binding of said antibody or antigen-binding fragment thereof,    wherein the binding of said antibody or antigen-binding fragment thereof to material in said sample is indicative of the presence of a mammalian CC-chemokine receptor 2 or portion of said receptor in said sample.    
     
     
         24 . A method according to  claim 23 , wherein the sample is a cellular fraction which, in normal individuals, comprises a mammalian CC-chemokine receptor 2 or portion of said receptor.  
     
     
         25 . A method of inhibiting a function associated with binding of a chemokine to a mammalian CC-chemokine receptor 2 or a functional portion of said receptor, comprising contacting a composition comprising the receptor or portion with an effective amount of an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody inhibits binding of said chemokine to mammalian CC-chemokine receptor 2 and inhibits one or more functions associated with binding of the ligand to the receptor.  
     
     
         26 . A method according to  claim 25  wherein the chemokine is selected from the group consisting of MCP-1, MCP-2, MCP-3, MCP-4 and combinations thereof.  
     
     
         27 . A method of detecting or identifying an agent which binds a mammalian CC-chemokine receptor 2 or ligand-binding variant thereof, comprising combining 
 a) an agent to be tested;    b) an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor; and    c) a composition comprising a mammalian CC-chemokine receptor 2 or a ligand-binding variant thereof,    under conditions suitable for binding of said antibody or antigen-binding fragment to said mammalian CC-chemokine receptor 2 or ligand-binding variant thereof, and detecting or measuring binding of said antibody or antigen-binding fragment to said mammalian CC-chemokine receptor 2 or ligand-binding variant thereof.    
     
     
         28 . A method according to  claim 27  wherein the formation of a complex between said antibody or antigen-binding fragment and said mammalian CC-chemokine receptor 2 or ligand-binding variant is monitored, and wherein a decrease in the amount of complex formed relative to a suitable control is indicative that the agent binds said receptor or ligand-binding variant thereof.  
     
     
         29 . A method according to  claim 27  wherein the composition comprising a mammalian CC-chemokine receptor 2 or a ligand-binding variant thereof is a cell bearing recombinant CC-chemokine receptor 2 or ligand-binding variant thereof.  
     
     
         30 . A method according to  claim 29 , wherein the composition comprising a mammalian CC-chemokine receptor 2 or a ligand-binding variant thereof is a membrane fraction of said cell bearing recombinant CC-chemokine receptor 2 or ligand-binding variant thereof.  
     
     
         31 . A method according to  claim 27  wherein the antibody or antigen-binding fragment thereof is labeled with a label selected from the group consisting of a radioisotope, spin label, antigen label, enzyme label, fluorescent group and chemiluminescent group.  
     
     
         32 . A method according to  claim 27  wherein the agent is an antibody having specificity for a mammalian CC-chemokine receptor 2 or antigen-binding fragment thereof.  
     
     
         33 . A method of inhibiting HIV infection in a patient, comprising administering to the patient a composition comprising an effective amount of an antibody or antigen-binding fragment thereof which binds to mammalian CC-chemokine receptor 2 and inhibits binding of HIV to the receptor.  
     
     
         34 . A method of inhibiting leukocyte trafficking in a patient, comprising administering to the patient a composition comprising an effective amount of an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2 and inhibits binding of a ligand to the receptor.  
     
     
         35 . A method according to  claim 34  wherein the ligand is a chemokine.  
     
     
         36 . A method according to  claim 35  wherein the chemokine is selected from the group consisting of MCP-1, MCP-2, MCP-3, MCP-4 and combinations of the foregoing.  
     
     
         37 . A composition comprising an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor, and an optional physiologically acceptable vehicle.  
     
     
         38 . An antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor with an IC 50  of less than about 1.0 μg/ml.  
     
     
         39 . An antibody or antigen-binding fragment thereof according to  claim 38  wherein the IC 50  is less than about 0.05 μg/ml.  
     
     
         40 . An antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor, and wherein the antibody or antigen-binding fragment binds the receptor with an affinity of at least about 0.1×10 −9  M.  
     
     
         41 . An antibody or antigen-binding fragment thereof according to  claim 40 , wherein the affinity is at least about 1×10 −9  M.  
     
     
         42 . An antibody or antigen-binding fragment thereof according to  claim 40 , wherein the affinity is at least about 3×10 −9  M.  
     
     
         43 . A method of treating a CC-chemokine receptor 2-mediated disorder in a patient, comprising administering to the patient an effective amount of an antibody or antigen-binding fragment thereof which binds to a mammalian CC-chemokine receptor 2, wherein said antibody or antigen-binding fragment thereof inhibits binding of a ligand to the receptor.