Humanized Anti-CCR2 Antibodies and Methods of Use Therefor

Abstract

The present invention relates to a humanized 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.

1 .- 38 . (canceled) 
     
     
         39 . A method of inhibiting a function associated with binding of a chemokine to a mammalian CC-chemokine receptor 2 (CCR2), comprising contacting a composition comprising the CCR2 with an effective amount of a humanized antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof inhibits binding of the chemokine to CCR2 and inhibits one or more functions associated with binding of the chemokine to CCR2. 
     
     
         40 . The method of  claim 39 , wherein the chemokine is selected from the group consisting of MCP-1, MCP-2, MCP-3, MCP-4 and combinations thereof. 
     
     
         41 . The method of  claim 40 , wherein said function is selected from the group consisting of:
 (a) signaling activity;   (b) stimulation of a cellular response; and   (c) combinations of (a) and (b).   
     
     
         42 . The method of  claim 41 , wherein said function is signaling activity and is selected from the group consisting of:
 (a) activation of a mammalian G protein;   (b) induction of a rapid and transient increase in the concentration of cytosolic free calcium [Ca 2+ ]I; and   (c) combinations of (a) and (b).   
     
     
         43 . The method of  claim 41 , wherein said function is stimulation of a cellular response and is selected from the group consisting of:
 (a) stimulation of chemotaxis;   (b) exocytosis;   (c) inflammatory mediator release by leukocytes;   (d) integrin activation;   (e) T cell activation;   (f) leukocyte degranulation; and   (g) combinations of (a), (b), (c), (d), (e) and (f).   
     
     
         44 . The method of  claim 39 , wherein the humanized antibody or antigen-binding fragment thereof comprises three complementarity determining region sequences of the light chain of the 1D9 antibody and a framework region sequence of the variable light chain of the HF21/28 antibody. 
     
     
         45 . The method of  claim 39 , wherein the humanized antibody or antigen-binding fragment thereof comprises three complementarity determining region sequence of the variable heavy chain of monoclonal antibody 1D9 and a framework region sequence of the variable heavy chain of the 4B4′CL antibody. 
     
     
         46 . The method of  claim 44 , wherein the humanized antibody or antigen-binding fragment thereof comprises three complementarity determining region sequences of the variable heavy chain of monoclonal antibody 1D9. 
     
     
         47 . The method of  claim 46 , wherein the humanized antibody or antigen-binding fragment thereof further comprises a framework region derived from the variable heavy chain of the 4B4′CL antibody. 
     
     
         48 . The method of  claim 39 , wherein the humanized antibody or antigen-binding fragment thereof comprises three complementarity determining region sequence of the variable light chain of monoclonal antibody 1D9, a framework region sequence of the variable light chain of the HF 21/28 antibody, three complementarity determining region sequence of the variable heavy chain of monoclonal antibody 1D9 and a framework region sequence of the variable heavy chain of the 4B4′CL antibody. 
     
     
         49 . The method of  claim 48 , wherein the humanized antibody or antigen-binding fragment thereof comprises a heavy chain constant region or portion thereof. 
     
     
         50 . The method of  claim 49 , wherein the human constant region or portion thereof is of the gamma type. 
     
     
         51 . The method of  claim 50 , wherein the human constant region or portion thereof is mutated to minimize binding to Fc receptors, the ability to fix complement or both. 
     
     
         52 . The method of  claim 48 , wherein the humanized antibody or antigen-binding fragment thereof, comprises a light chain constant region. 
     
     
         53 . The method of  claim 52 , wherein the human light chain constant region is of the kappa type. 
     
     
         54 . The method of  claim 48 , wherein the light chain variable region of the humanized antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:12. 
     
     
         55 . The method of  claim 48 , wherein the heavy chain variable region of the humanized antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:17. 
     
     
         56 . The method of  claim 48 , wherein the light chain variable region of the humanized antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO: 12, and the heavy chain variable region of the humanized antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:17. 
     
     
         57 . The method of  claim 56 , wherein the humanized antibody or antigen-binding fragment thereof, comprises a heavy chain constant region or portion thereof. 
     
     
         58 . The method of  claim 57 , wherein the human constant region or portion thereof is of the gamma type. 
     
     
         59 . The method of  claim 58 , wherein the human constant region or portion thereof is mutated to minimize binding to Fc receptors, the ability to fix complement or both. 
     
     
         60 . The method of  claim 56 , wherein the humanized antibody or antigen-binding fragment thereof, comprises a light chain constant region. 
     
     
         61 . The method of  claim 60 , wherein the human light chain constant region is of the kappa type.