Heterologous antibodies which bind human cd4

Abstract

The invention relates to transgenic non-human animals capable of producing high affinity human sequence antibodies. The invention is also directed to human sequence antibodies specific for human antigens, such as, human CD4. The invention is also directed to methods for producing human sequence antibodies.

1 . An isolated nucleic acid encoding a heavy chain variable region of an antibody, or antigen binding portion thereof, that binds to human CD4, comprising nucleotide sequences encoding the heavy chain CDR1, CDR2, and CDR3 regions,
 wherein the nucleotide sequence encoding the heavy chain CDR3 region comprises nucleotides 349-369 of SEQ ID NO:205 or nucleotides 361-381 of SEQ ID NO:219, the nucleotide sequence encoding the heavy chain CDR2 region comprises nucleotides 205-252 of SEQ ID NO:205 or nucleotides 217-264 of SEQ ID NO:219, and the nucleotide sequence encoding the heavy chain CDR1 region comprises nucleotides 148-162 of SEQ ID NO:205 or nucleotides 160-174 of SEQ ID NO:219,   or sequences having at least 90% identity thereto.   
     
     
         2 . An isolated nucleic acid encoding a light chain variable region of an antibody, or antigen binding portion thereof, that binds to human CD4, comprising nucleotide sequences encoding the heavy chain CDR1, CDR2, and CDR3 regions,
 wherein the nucleotide sequence encoding the light chain CDR3 region comprises nucleotides 331-357 of SEQ ID NO:206 or nucleotides 337-363 of SEQ ID NO:220, the nucleotide sequence encoding the light chain CDR2 region comprises nucleotides 214-234 of SEQ ID NO:206 or nucleotides 220-240 of SEQ ID NO:220, and the nucleotide sequence encoding the light chain CDR1 region comprises nucleotides 136-168 of SEQ ID NO:206 and nucleotides 142-174 of SEQ ID NO:220,   or sequences having at least 90% identity thereto.   
     
     
         3 . A vector comprising the nucleic acid of  claim 1 . 
     
     
         4 . A cell comprising the vector of  claim 3 . 
     
     
         5 . A method of producing an isolated human monoclonal antibody, or antigen binding portion thereof, which binds to human CD4, comprising:
 immunizing a transgenic non-human animal having a genome comprising a human heavy chain transgene and a human light chain transgene with human CD4 or a cell expressing human CD4, such that antibodies are produced by B cells of the animal,   isolating B cells of the animal;   fusing the B cells with myeloma cells to form immortal, hybridoma cells that secrete human monoclonal antibodies which bind to human CD4; and   isolating the human monoclonal antibodies specific for human CD4 from the culture supernatant of the hybridoma, or from a transfectoma derived from such hybridoma, wherein the antibody comprises a variable region encoded by the nucleic acid of  claim 1 .   
     
     
         6 . A method of treating a disease associated with cells which express CD4, comprising administering to a subject in need thereof a monoclonal antibody, or antigen binding portion thereof, wherein the antibody comprises a variable region encoded by the nucleic acid of  claim 1 . 
     
     
         7 . The method of  claim 6 , wherein the disease is selected from the group consisting of an autoimmune disease, an inflammatory disease, reperfusion injury, an infectious disease, a blood disorder, and transplant rejection. 
     
     
         8 . The method of  claim 7 , wherein the disease is an autoimmune disease. 
     
     
         9 . The method of  claim 8 , wherein the disease is rheumatoid arthritis. 
     
     
         10 . The method of  claim 6 , wherein the antibody, or portion thereof, has at least one of the following properties:
 the antibody is an IgG1 antibody;   the antibody comprises a variable region from an IgG1 heavy chain and a variable region from a kappa light chain;   the antibody dissociates from human CD4 with a dissociation rate constant (Kdiss) of about 4.2×10 −5  S −1  or less;   the antibody binds to human CD4 with an association rate constant (K ass ) of at least about 7.4×10 4  M −1  S −1 ;   the antibody binds to human CD4 with an affinity constant (Ka) of at least about 1.0×10 9  M −1 ;   the antibody inhibits a mixed leukocyte reaction; or   the antibody binds CD4 from non-human primates.   
     
     
         11 . The method of  claim 6 , wherein the antibody, or antigen binding portion thereof, is a Fab fragment or a single chain antibody. 
     
     
         12 . The method of  claim 6 , wherein the antibody, or portion thereof, is linked to a therapeutic agent. 
     
     
         13 . The method of  claim 12 , wherein the therapeutic agent is a radiolabel or a toxin. 
     
     
         14 . A method of diagnosing a disease associated with cells which express CD4 comprising:
 (i) contacting a biological sample from a subject and a control sample with a monoclonal antibody, or antigen binding portion thereof, under conditions that allow for formation of a complex between the antibody, or antigen binding portion thereof, and CD4, and   (ii) detecting the difference in complex formation between the two samples, wherein a difference in the complex formation between the two samples indicates that the subject has, or is susceptible to developing, the disease,   wherein the antibody comprises a variable region encoded by the nucleic acid of  claim 1 .   
     
     
         15 . The method of  claim 14 , wherein the disease is selected from the group consisting of an autoimmune disease, an inflammatory disease, reperfusion injury, an infectious disease, a blood disorder, and transplant rejection. 
     
     
         16 . The method of  claim 15 , wherein the disease is an autoimmune disease. 
     
     
         17 . The method of  claim 16 , wherein the disease is rheumatoid arthritis. 
     
     
         18 . A hybridoma that produces a monoclonal antibody that specifically binds human CD4, wherein the antibody comprises heavy and light chain CDR1, CDR2, and/or CDR3 regions and wherein:
 (a) the heavy chain CDR3 region is encoded by nucleotides 349-369 of SEQ ID NO:205 or nucleotides 361-381 of SEQ ID NO:219 and the light chain CDR3 region is encoded by nucleotides 331-357 of SEQ ID NO:206 or nucleotides 337-363 of SEQ ID NO:220;   (b) the heavy chain CDR2 region is encoded by nucleotides 205-252 of SEQ ID NO:205 or nucleotides 217-264 of SEQ ID NO:219 and the light chain variable region CDR2 region is encoded by nucleotides 214-234 of SEQ ID NO:206 or nucleotides 220-240 of SEQ ID NO:220; and   (c) the heavy chain CDR1 region is encoded by nucleotides 148-162 of SEQ ID NO:205 or nucleotides 160-174 of SEQ ID NO:219 and the light chain variable region CDR1 region is encoded by nucleotides 136-168 of SEQ ID NO:206 and nucleotides 142-174 of SEQ ID NO:220.   
     
     
         19 . A vector comprising the nucleic acid of  claim 2 . 
     
     
         20 . A cell comprising the vector of  claim 19 .