Immunogenetic restriction on elicitation of antibodies

Abstract

The present invention provides structural determinants important for binding to the stem domain of the HA protein of influenza virus, and methods of use thereof for production of high affinity neutralizing influenza virus antibodies based upon these determinants. The present invention further provides tools for determining the efficacy of an influenza virus vaccine. The present invention further provides a molecular signature useful for determining the efficacy of an influenza virus vaccine in a subject, or for predicting prior immunologic exposure or antigen responsiveness to vaccine or influenza virus infection.

1 .- 38 . (canceled) 
     
     
         39 . An isolated monoclonal antibody comprising in the VH domain a methionine, isoleucine, valine, or leucine at position 53; a phenylalanine at position 54; a tyrosine at position 97, 98 or 99; wherein the positions of the amino acids are numbered based on the Kabat numbering system; and wherein the VH domain is encoded by the immunoglobulin VH1-69 germline gene. 
     
     
         40 . The antibody of  claim 39 , further comprising in the VH domain a serine at position 24; a valine a position 27; an isoleucine or proline at position 28; a serine at position 29; an arginine at position 30; a valine at position 34; a serine at position 52; a glycine at position 52a; a lysine at position 58; a glutamine at position 73; a phenylalanine at position 74; or any combination thereof. 
     
     
         41 . The antibody according to  claim 39 , wherein the antibody binds to the stem region of HA protein of an influenza virus. 
     
     
         42 . The antibody of  claim 41 , wherein the antibody neutralizes an influenza virus. 
     
     
         43 . The antibody of  claim 42 , wherein said influenza virus is an influenza A virus. 
     
     
         44 . The antibody according to  claim 43 , wherein said influenza virus is a Group I influenza virus. 
     
     
         45 . The antibody according to  claim 39 , wherein the antibody is a single chain Fv antibody, an F ab  fragment, an F ab′  fragment, or an F (ab′)2  fragment. 
     
     
         46 . The antibody according to  claim 39 , wherein the antibody is linked to a therapeutic agent. 
     
     
         47 . The antibody of  claim 46 , wherein said therapeutic agent is a toxin, a radiolabel, a siRNA, a small molecule, or a cytokine. 
     
     
         48 . The antibody according to  claim 39 , wherein in the VH domain position 24 has been mutated a serine; position 27 has been mutated to a valine; position 28 has been mutated to an isoleucine; position 29 has been mutated to a serine; position 30 has been mutated to arginine; position 34 has been mutated to a valine; position 52 has been mutated to a serine; position 52a has been mutated to a glycine or alanine; position 58 has been mutated to lysine; position 73 has been mutated to glutamine; position 74 has been mutated to a phenylalanine; or any combination thereof. 
     
     
         49 . The antibody according to  claim 48 , wherein in the VH domain position 24 has been mutated from alanine to serine, the amino acid at position 27 has been mutated from glycine to valine, the amino acid at position 28 has been mutated from threonine to isoleucine or proline, the amino acid at position 29 has been mutated from phenylalanine to serine, the amino acid at position 30 has been mutated from serine to arginine, the amino acid at position 34 has been mutated from isoleucine to valine, the amino acid at position 52 has been mutated from isoleucine to serine, the amino acid at position 52a has been mutated from proline to glycine or alanine, the amino acid at position 58 has been mutated from asparagine to lysine, the amino acid at position 73 has been mutated from glutamate to glutamine, has been mutated the amino acid at position 74 from serine to phenylalanine, or any combination thereof; wherein the positions of the amino acids are numbered based on the Kabat numbering system. 
     
     
         50 . The antibody according to  claim 48 , wherein in the VH domain position 52 has been mutated to a serine and position 52a has been mutated to a glycine. 
     
     
         51 . The antibody according to  claim 48 , wherein in the VH domain position 73 has been mutated to a glutamine and position 74 has been mutated to a phenylalanine. 
     
     
         52 . The antibody according to  claim 48 , wherein in the VH domain the antibody comprises a methionine or isoleucine at position 53; a phenylalanine at position 54; and a tyrosine at position 97, 98, 99. 
     
     
         53 . A cell producing the antibody according to  claim 39 . 
     
     
         54 . A composition comprising the antibody of  claim 39  and a carrier. 
     
     
         55 . A method of preventing or treating a disease or disorder caused by an influenza virus, the method comprising administering to a person at risk of suffering from said disease or disorder a therapeutically effective amount of the monoclonal antibody of  claim 39 . 
     
     
         56 . The method of  claim 55 , wherein said influenza virus is an influenza A virus. 
     
     
         57 . The method of  claim 56 , wherein said influenza virus is a Group I influenza virus.