Affinity matured anti-ccr4 humanized monoclonal antibodies and methods of use

Abstract

The present invention provides affinity matured humanized monoclonal antibodies, bi-specific antibodies, antibody conjugates, and fusion proteins that bind to the chemokine receptor CCR4. This antibody is derived from mAb 1567 and recognizes the same epitope. Binding of the antibodies disclosed herein to CCR4 inhibits ligand-mediated activities and is used to treat symptoms of cancer. Moreover, the antibody is used in combination with vaccines to suppress the activity of regulatory T cells.

1 . An isolated humanized monoclonal antibody having:
 a. a heavy chain with three CDRs comprising the amino acid sequences GYTFASQW (SEQ ID NO:22), INPGNVNT (SEQ ID NO:27), and STWYRPLDY (SEQ ID NO:30) respectively and a light chain with three CDRs comprising the amino acid sequences QSILYSSNQKNY (SEQ ID NO:26), WASTRE (SEQ ID NO:28), and HQYISSYT (SEQ ID NO:35) respectively;   b. a heavy chain with three CDRs comprising the amino acid sequences GYTFASSW (SEQ ID NO:23), INPGNVNT (SEQ ID NO:27), and STWYRPNDY (SEQ ID NO:31) respectively and a light chain with three CDRs comprising the amino acid sequences QSILYSSNQKNY (SEQ ID NO:26), WASTRE (SEQ ID NO:28), and HQYKSSYT (SEQ ID NO:36) respectively;   c. a heavy chain with three CDRs comprising the amino acid sequences GYTFASSW (SEQ ID NO:23), INPGNVNT (SEQ ID NO:27), and TTRYRPLDY (SEQ ID NO:32) respectively and a light chain with three CDRs comprising the amino acid sequences QSILYSSNQKNY (SEQ ID NO:26), WASTRE (SEQ ID NO:28), and HQYRSSYT (SEQ ID NO:37) respectively;   d. a heavy chain with three CDRs comprising the amino acid sequences GYTFASQY (SEQ ID NO:24), INPGNVNT (SEQ ID NO:27), and LTYYRPPDY (SEQ ID NO:33) respectively and a light chain with three CDRs comprising the amino acid sequences QSILYSSNQKNY (SEQ ID NO:26), WASTRE (SEQ ID NO:28), and HQYYSSYT (SEQ ID NO:38) respectively; or   e. a heavy chain with three CDRs comprising the amino acid sequences GYTFASAW (SEQ ID NO:25), INPGNVNT (SEQ ID NO:27), and STYYRPLDY (SEQ ID NO:29) respectively and a light chain with three CDRs comprising the amino acid sequences QSILYSSNQKNY (SEQ ID NO:26), WASTRE (SEQ ID NO:28), and HQYMSSYT (SEQ ID NO:39) respectively;   
       wherein said antibody binds human CCR4. 
     
     
         2 . The antibody of  claim 1 , wherein said antibody is monovalent or bivalent. 
     
     
         3 . The antibody of  claim 1 , wherein said antibody is a single chain antibody. 
     
     
         4 - 12 . (canceled) 
     
     
         13 . A method of selectively killing a tumor cell comprising contacting said cell with the antibody of  claim 1 . 
     
     
         14 . The method of  claim 13 , wherein said selective killing occurs by antibody-dependent cellular toxicity (ADCC), complement-dependent cytotoxicity (CDC), antibody dependent cellular phagocytosis (ADCP). 
     
     
         15 . The method of  claim 14 , wherein said tumor cell expresses CCR4. 
     
     
         16 . (canceled) 
     
     
         17 . A method of decreasing suppressor T-cell activity comprising contacting said cell with the antibody of  claim 1 . 
     
     
         18 . A method of augmenting an immune response to an antigen comprising contacting a T cell with the antibody of  claim 1 . 
     
     
         19 .- 23 . (canceled) 
     
     
         24 . A method of increasing T cell proliferation comprising contacting a T cell with the antibody of  claim 1 . 
     
     
         25 . A method of reversing regulatory T cell-mediated suppression of effector T cell proliferation comprising contacting a T cell with the antibody of  claim 1 . 
     
     
         26 . A method of increasing cytokine production or secretion comprising contacting a T cell with the antibody of  claim 1 . 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . A method of increasing vaccine efficiency comprising administering to a subject an antibody according to  claim 1  and a vaccine. 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . A method of treating or alleviating a symptom of cancer, comprising administering to a subject in need thereof a composition comprising an antibody according to  claim 1 . 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . A method of treating or alleviating a symptom of an autoimmune disease comprising administering to a subject in need thereof a composition comprising an antibody according to  claim 1 , wherein said antibody is linked to a regulatory T-cell expansion agent. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . A method of decreasing regulatory T-cell activity comprising contacting a regulatory T-cell with an antibody according to  claim 1 , wherein said antibody is linked to toxin. 
     
     
         44 . (canceled) 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . A nucleic acid sequence comprising the nucleic acid sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17 or 19. 
     
     
         49 . (canceled) 
     
     
         50 . (canceled) 
     
     
         51 . A vector comprising the nucleic acid of  claim 48 . 
     
     
         52 . A cell comprising the vector of  claim 51 .