Interleukin-10 antibodies

Abstract

The methods and compositions provided herein relate generally to IL-10 specific antibodies and uses thereof. More specifically, compositions of humanized IL-10 specific antibodies and methods to use such antibodies in modulating the biological activity of IL-10, particularly in autoimmune disorders and pathogen-mediated immunopathology.

1 . A humanized recombinant antibody molecule that binds IL-10 or binding fragment thereof, comprising: 
 at least one antibody light chain variable region, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected from the group consisting of SEQ ID NO:1 at CDR1, SEQ ID NO:2 at CDR2, and SEQ ID NO:3 at CDR3; and a framework region, wherein the amino acid sequence of framework region is all or substantially all of a human immunoglobin amino acid sequence; and    at least one antibody heavy chain variable region, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected from the group consisting of SEQ ID NO:6 at CDR1, SEQ ID NO:7 at CDR2, and SEQ ID NO:8 at CDR3; and a framework region, wherein the amino acid sequence of framework region is all or substantially all of a human immunoglobin amino acid sequence.    
     
     
         2 . The antibody of  claim 1 , further comprising a heavy chain constant region, wherein the heavy chain constant region comprises a γ1, γ2, γ3, or γ4 human heavy chain constant region or a variant thereof.  
     
     
         3 . The antibody of  claim 1 , further comprising a light chain constant region, wherein the light chain constant region comprises a lambda or a kappa human light chain constant region.  
     
     
         4 . The antibody of  claim 1 , wherein the binding fragment is an antibody fragment selected from the group consisting of Fab, Fab′, Fab′-SH, Fv, scFv, F(ab′) 2 , and a diabody.  
     
     
         5 . The antibody of  claim 1 , wherein the antibody light chain, or binding fragment thereof, comprises a polypeptide having a variable region of SEQ ID NO:4.  
     
     
         6 . The antibody of  claim 1 , wherein the antibody light chain, or binding fragment thereof, comprises a polypeptide having a variable region and a constant region of SEQ ID NO:5.  
     
     
         7 . The antibody of  claim 1 , wherein the antibody heavy chain, or binding fragment thereof, comprises a polypeptide having a variable region of SEQ ID NO:9.  
     
     
         8 . The antibody of  claim 1 , wherein the antibody heavy chain, or binding fragment thereof, comprises a polypeptide having a variable region and a constant region of SEQ ID NO:10.  
     
     
         9 . A chimeric recombinant antibody molecule that binds IL-10 or binding fragment thereof, comprising: 
 at least one antibody light chain variable region, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected from the group of SEQ ID NO:1 at CDR1, SEQ ID NO:2 at CDR2, and SEQ ID NO:3 at CDR3; and    at least one antibody heavy chain variable region, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected from the group of SEQ ID NO:6 at CDR1, SEQ ID NO:7 at CDR2, and SEQ ID NO:8 at CDR3.    
     
     
         10 . A humanized recombinant antibody molecule that binds IL-10, or binding fragment thereof, comprising: 
 at least one antibody light chain, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected from the group consisting of SEQ ID NO:1 at CDR1, SEQ ID NO:12 at CDR2, and SEQ ID NO:13 at CDR3; and a framework region, wherein the amino acid sequence of framework region is all or substantially all of a human immunoglobin amino acid sequence; and    at least one antibody heavy chain, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence selected of SEQ ID NO:15 at CDR1, SEQ ID NO:16 at CDR2, and SEQ ID NO:17 at CDR3; and a framework region, wherein the amino acid sequence of framework region is all or substantially all of a human immunoglobin amino acid sequence.    
     
     
         11 . The antibody of  claim 10 , further comprising a heavy chain constant region, wherein the heavy chain constant region comprises a γ1, γ2, γ3, or γ4 human heavy chain constant region or a variant thereof.  
     
     
         12 . The antibody of  claim 10 , further comprising a light chain constant region, wherein the light chain constant region comprises a lambda or a kappa human light chain constant region.  
     
     
         13 . The antibody of  claim 10 , wherein the binding fragment is an antibody fragment selected from the group consisting of Fab, Fab′, Fab′-SH, Fv, scFv, F(ab′) 2 , and diabody.  
     
     
         14 . The antibody of  claim 10 , wherein the antibody light chain, or binding fragment thereof, comprises a polypeptide having a variable region and a constant region of SEQ ID NO:14.  
     
     
         15 . The antibody of  claim 10 , wherein the antibody heavy chain, or binding fragment thereof, comprises a polypeptide having a variable region and a constant region of SEQ ID NO:18.  
     
     
         16 . A chimeric recombinant antibody molecule that binds IL-10, or binding fragment thereof, comprising: 
 at least one antibody light chain, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence of SEQ ID NO:11 at CDR1, SEQ ID NO:12 at CDR2, and SEQ ID NO:13 at CDR3; and    at least one antibody heavy chain, or binding fragment thereof, comprising a polypeptide having at least one amino acid sequence of SEQ ID NO:15 at CDR1, SEQ ID NO:16 at CDR2, and SEQ ID NO:17 at CDR3.    
     
     
         17 . A method of suppressing an immune response in a human subject comprising administering to a subject in need thereof an antibody specific for IL-10, or a binding fragment thereof, in an amount effective to block the biological activity of IL-10, wherein the antibody is the antibody of  claim 1  or  9 .  
     
     
         18 . The method of  claim 17 , wherein the immune response is a humoral response.  
     
     
         19 . The method of  claim 17 , wherein the subject has systemic lupus erythematosus.  
     
     
         20 . The method of  claim 17 , wherein the subject has immune thrombocytopenic purpura (ITC).  
     
     
         21 . The method of  claim 17 , wherein the subject has lupus nephritis.  
     
     
         22 . The method of  claim 17 , wherein the subject has HIV.  
     
     
         23 . The method of  claim 17 , wherein the subject has hepatitis C.  
     
     
         24 . The method of  claim 17 , further comprising administering an immunosuppressive agent.  
     
     
         25 . A method of suppressing an immune response in a human subject comprising administering to a subject in need thereof, an antibody specific for IL-10, or a binding fragment thereof, in an amount effective to block the biological activity of IL-10, wherein the antibody is the antibody of  claim 10  or  16 .  
     
     
         26 . The method of  claim 25 , wherein the immune response is a humoral response.  
     
     
         27 . The method of  claim 25 , wherein the subject has systemic lupus erythematosus.  
     
     
         28 . The method of  claim 25 , wherein the subject has immune thrombocytopenic purpura (ITC).  
     
     
         29 . The method of  claim 25 , wherein the subject has lupus nephritis.  
     
     
         30 . The method of  claim 25 , wherein the subject has HIV.  
     
     
         31 . The method of  claim 25 , wherein the subject has hepatitis C.  
     
     
         32 . The method of  claim 25 , further comprising administering an immunosuppressive agent.  
     
     
         33 . A composition comprising an antibody, or binding fragment thereof, in combination with a pharmaceutically acceptable carrier or diluent, wherein the antibody is the antibody of  claim 1  or  9 .  
     
     
         34 . The composition of  claim 33 , further comprising an immunosuppressive agent.  
     
     
         35 . A composition comprising an antibody, or binding fragment thereof, in combination with a pharmaceutically acceptable carrier or diluent, wherein the antibody is the antibody of  claim 10  or  16 .  
     
     
         36 . The composition of  claim 35 , further comprising an immunosuppressive agent.  
     
     
         37 . An isolated nucleic acid encoding the polypeptide of  claim 1  or  9 .  
     
     
         38 . An isolated nucleic acid encoding the polypeptide of  claim 10  or  16 .  
     
     
         39 . An expression vector comprising the nucleic acid sequence of  claim 37  operably linked to control sequences recognized by a host cell transfected with the vector.  
     
     
         40 . A host cell comprising the vector of  claim 38 .  
     
     
         41 . A method of producing a polypeptide, comprising culturing the host cell of  claim 40  under conditions wherein the nucleic acid sequence is expressed, thereby producing the polypeptide, and recovering the polypeptide from the host cell.  
     
     
         42 . An expression vector comprising the nucleic acid sequence of  claim 38  operably linked to control sequences recognized by a host cell transfected with the vector.  
     
     
         43 . A host cell comprising the vector of  claim 42 .  
     
     
         44 . A method of producing a polypeptide, comprising culturing the host cell of  claim 43  under conditions wherein the nucleic acid sequence is expressed, thereby producing the polypeptide, and recovering the polypeptide from the host cell.  
     
     
         45 . An isolated nucleic acid sequence encoding an antibody specific for IL-10 comprising a light chain having the nucleic acid sequence of SEQ ID NO:21 and a heavy chain having the nucleic acid sequence of SEQ ID NO:22.  
     
     
         46 . The nucleic acid of  claim 45 , wherein the light chain has an American Type Culture Collection (ATCC) deposit number of PTA-5923 and the heavy chain has an ATCC deposit number of PTA-5922.  
     
     
         47 . An isolated nucleic acid sequence encoding an antibody specific for IL-10 comprising a light chain having the nucleic acid sequence of SEQ ID NO:23 and a heavy chain having the nucleic acid sequence of SEQ ID NO:24.  
     
     
         48 . The nucleic acid of  claim 47 , wherein light chain has an ATCC deposit number of PTA-5927 and the heavy chain has an ATCC deposit number of PTA-5926.  
     
     
         49 . An isolated nucleic acid sequence encoding a binding fragment of the antibody encoded by the nucleic acid sequence of  claim 45 .  
     
     
         50 . An isolated nucleic acid sequence encoding a binding fragment of the antibody encoded by the nucleic acid sequence of  claim 47 .  
     
     
         51 . The nucleic acid of  claim 49  or  50 , wherein the binding fragment is an antibody fragment selected from the group consisting of Fab, Fab′, Fab′-SH, Fv, scFv, F(ab′) 2 , and a diabody.  
     
     
         52 . A method to identify an acceptor germline sequence for a humanized antibody, which method comprises the steps of: 
 a) identifying a non-human antibody that has the desired biological activity;    b) determining the amino acid sequence of a non-human antibody V H  and V L  domains; and    c) comparing the nonhuman antibody sequence to a group of human germline sequences, wherein the comparison comprises the substeps of: 
 1) assigning the sequence of non-human V H  and V L  domain sequences residue numbers;  
 2) delineating the CDR and FR regions in the sequence;  
 3) assigning a predetermined numerical score at each residue position for which the non-human and human germline sequences are identical; and  
 4) totaling all of the residue scores to generate a total score for each human germline sequence; and  
   d) identifying the human germline sequence with the highest total residue score as the acceptor germline sequence.    
     
     
         53 . The method of  claim 52 , further comprising the substeps of: 
 a) assigning a numerical score of 1 for each residue position for which the non-human and human germline sequences are identical that was not scored in substep (3) to germline sequences with identical total residue scores after substep (4);    b) totaling all of the residue scores to generate a total score for each human germline sequence.    
     
     
         54 . The method of  claim 52 , wherein the V H  region is scored as in Table 2.  
     
     
         55 . The method of  claim 52 , wherein the V L  region is scored as in Table 3.  
     
     
         56 . The method of  claim 52 , wherein the non-human antibody is specific for IL-10 and inhibits the biological activity of IL-10.  
     
     
         57 . An antibody generated by the method of  claim 52.