Monoclonal antibodies against influenza virus generated by cyclical administration and uses thereof

Abstract

Provided herein are methods of producing neutralizing monoclonal antibodies, by cyclical immunization, that cross-react with strains of Influenza virus of the same subtype or different subtypes. Also provided herein are compositions comprising such antibodies and methods of using such antibodies to diagnose, prevent or treat Influenza virus disease.

1 .- 50 . (canceled) 
     
     
         51 . A method for preventing influenza virus disease in a human subject in need thereof, comprising administering to the human subject a nucleic acid encoding an antibody by way of gene therapy, wherein the antibody binds to influenza virus hemagglutinin and comprises:
 (a) a heavy chain variable region comprising a complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO:38, a CDR2 having the amino acid sequence of SEQ ID NO:39, and a CDR3 having the amino acid sequence of SEQ ID NO:40; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:46, a CDR2 having the amino acid sequence of SEQ ID NO:47, and a CDR3 having the amino acid sequence of SEQ ID NO:48;   (b) a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:70, a CDR2 having the amino acid sequence of SEQ ID NO:71, and a CDR3 having the amino acid sequence of SEQ ID NO:72; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:78, a CDR2 having the amino acid sequence of SEQ ID NO:79, and a CDR3 having the amino acid sequence of SEQ ID NO:80;   (c) a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:92, a CDR2 having the amino acid sequence of SEQ ID NO:93, and a CDR3 having the amino acid sequence of SEQ ID NO:94; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 100, a CDR2 having the amino acid sequence of SEQ ID NO: 101, and a CDR3 having the amino acid sequence of SEQ ID NO: 102; or   (d) a heavy chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4; and a light chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4.   
     
     
         52 . The method of  claim 51 , wherein the antibody comprises a heavy chain variable region comprising a complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO:38, a CDR2 having the amino acid sequence of SEQ ID NO:39, and a CDR3 having the amino acid sequence of SEQ ID NO:40; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:46, a CDR2 having the amino acid sequence of SEQ ID NO:47, and a CDR3 having the amino acid sequence of SEQ ID NO:48. 
     
     
         53 . The method of  claim 51 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:70, a CDR2 having the amino acid sequence of SEQ ID NO:71, and a CDR3 having the amino acid sequence of SEQ ID NO:72; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:78, a CDR2 having the amino acid sequence of SEQ ID NO:79, and a CDR3 having the amino acid sequence of SEQ ID NO:80. 
     
     
         54 . The method of  claim 51 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:92, a CDR2 having the amino acid sequence of SEQ ID NO:93, and a CDR3 having the amino acid sequence of SEQ ID NO:94; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 100, a CDR2 having the amino acid sequence of SEQ ID NO: 101, and a CDR3 having the amino acid sequence of SEQ ID NO: 102. 
     
     
         55 . The method of  claim 51 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4; and a light chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4. 
     
     
         56 . A method for transferring/conferring passive immunity to influenza virus in a human subject in need thereof, comprising administering to the human subject a nucleic acid encoding an antibody by way of gene therapy, wherein the antibody binds to influenza virus hemagglutinin and comprises:
 (a) a heavy chain variable region comprising a complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO:38, a CDR2 having the amino acid sequence of SEQ ID NO:39, and a CDR3 having the amino acid sequence of SEQ ID NO:40; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:46, a CDR2 having the amino acid sequence of SEQ ID NO:47, and a CDR3 having the amino acid sequence of SEQ ID NO:48;   (b) a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:70, a CDR2 having the amino acid sequence of SEQ ID NO:71, and a CDR3 having the amino acid sequence of SEQ ID NO:72; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:78, a CDR2 having the amino acid sequence of SEQ ID NO:79, and a CDR3 having the amino acid sequence of SEQ ID NO:80;   (c) a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:92, a CDR2 having the amino acid sequence of SEQ ID NO:93, and a CDR3 having the amino acid sequence of SEQ ID NO:94; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 100, a CDR2 having the amino acid sequence of SEQ ID NO: 101, and a CDR3 having the amino acid sequence of SEQ ID NO: 102; or   (d) a heavy chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4; and a light chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4.   
     
     
         57 . The method of  claim 56 , wherein the antibody comprises a heavy chain variable region comprising a complementarity determining region (CDR)l having the amino acid sequence of SEQ ID NO:38, a CDR2 having the amino acid sequence of SEQ ID NO:39, and a CDR3 having the amino acid sequence of SEQ ID NO:40; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:46, a CDR2 having the amino acid sequence of SEQ ID NO:47, and a CDR3 having the amino acid sequence of SEQ ID NO:48. 
     
     
         58 . The method of  claim 56 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:70, a CDR2 having the amino acid sequence of SEQ ID NO:71, and a CDR3 having the amino acid sequence of SEQ ID NO:72; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:78, a CDR2 having the amino acid sequence of SEQ ID NO:79, and a CDR3 having the amino acid sequence of SEQ ID NO:80. 
     
     
         59 . The method of  claim 56 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO:92, a CDR2 having the amino acid sequence of SEQ ID NO:93, and a CDR3 having the amino acid sequence of SEQ ID NO:94; and a light chain variable region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 100, a CDR2 having the amino acid sequence of SEQ ID NO: 101, and a CDR3 having the amino acid sequence of SEQ ID NO: 102. 
     
     
         60 . The method of  claim 56 , wherein the antibody comprises a heavy chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4; and a light chain variable region comprising a CDR1 of the antibody 39A4, a CDR2 of the antibody 39A4, and a CDR3 of the antibody 39A4. 
     
     
         61 . A method for preventing influenza virus disease in a human subject in need thereof, comprising administering to the human subject a nucleic acid encoding an antibody by way of gene therapy, wherein the antibody binds to influenza virus hemagglutinin and neutralizes an influenza virus of the H1 subtype and an influenza virus of the H3 subtype. 
     
     
         62 . A method for transferring/conferring passive immunity to influenza in a human subject in need thereof, comprising administering to the human subject a nucleic acid encoding an antibody by way of gene therapy, wherein the antibody binds to influenza virus hemagglutinin and neutralizes an influenza virus of the HI subtype and an influenza virus of the H3 subtype.