Restricted immunoglobulin heavy chain mice

Abstract

Mice having a restricted immunoglobulin heavy chain locus are provided, wherein the locus is characterized by a single polymorphic human VH gene segment, a plurality of human DH gene segments and a plurality of JH gene segments. Methods for making antibody sequences that bind an antigen (e.g., a viral antigen) are provided, comprising immunizing a mouse with an antigen of interest, wherein the mouse comprises a single human VH gene segment, a plurality of human DH gene segments and a plurality of JH gene segments, at the endogenous immunoglobulin heavy chain locus.

1 . A rat or mouse having a restricted immunoglobulin heavy chain locus characterized by the presence of a single human V H  gene segment, one or more human D H  gene segments, and one or more human J H  gene segments. 
     
     
         2 . The rat or mouse of  claim 1 , wherein the mouse comprises a deletion of all or substantially all endogenous V H , D H , and J H  gene segments. 
     
     
         3 . The mouse of  claim 1 , wherein the single human V H  gene segment is selected from V H 1-2, V H 1-69, V H 2-26, V H 2-70, V H 3-23. 
     
     
         4 .- 5 . (canceled) 
     
     
         6 . The rat or mouse of  claim 1 , wherein the single human V H  gene segment is operably linked to a human or non-human immunoglobulin constant region gene. 
     
     
         7 . The rat or mouse of  claim 6 , wherein the single human V H  gene segment is operably linked to a mouse, rat, or human constant region gene. 
     
     
         8 . The rat or mouse of  claim 1 , further comprising a one or more human immunoglobulin V L  gene segments operably linked to one or more human J L  gene segments. 
     
     
         9 . The rat or mouse of  claim 8 , wherein the one or more human V L  gene segments and/or the one or more human J L  gene segments are selected from human κ and human λ gene segments. 
     
     
         10 . The rat or mouse of  claim 8 , wherein the one or more human immunoglobulin V L  gene segments and the one or more human J L  gene segments are operably linked to a non-human light chain constant gene. 
     
     
         11 . The rat or mouse of  claim 10 , wherein the non-human light chain constant gene is selected from a mouse κ or λ constant region gene. 
     
     
         12 . A mouse comprising, in its germline, a replacement at an endogenous immunoglobulin heavy chain locus of all or substantially all endogenous V H , D H , and J H  gene segments with single human V H  gene segment and/or polymorphic variants thereof, one or more human D H  gene segments and one or more human J H  gene segments. 
     
     
         13 . The mouse of  claim 12 , further comprising a replacement at an endogenous immunoglobulin light chain locus of all or substantially all endogenous V L  and J L  gene segments with one or more human V L  and one or more human J L  gene segments. 
     
     
         14 . (canceled) 
     
     
         15 . A cell or tissue derived from the rat or mouse of  claim 1 . 
     
     
         16 . A method of making a nucleic acid sequence encoding a human V H  domain, comprising
 (a) immunizing a rat or mouse of  claim 1  with an antigen of interest;   (b) allowing said mouse to mount an immune response to the antigen of interest; and,   (c) obtaining a nucleic acid sequence encoding a human V H  domain from said mouse.   
     
     
         17 .- 19 . (canceled) 
     
     
         20 . The method of  claim 16 , wherein the immunoglobulin V H  region sequence is at least 95% identical to SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66 or SEQ ID NO: 68. 
     
     
         21 . The method of  claim 16 , wherein the immunoglobulin V H  region sequence comprises SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68. or a polymorphic variant thereof. 
     
     
         22 . The method of  claim 16 , wherein the immunoglobulin V H  region sequence encodes a protein that is at least 95% identical with SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67 or SEQ ID NO: 69. 
     
     
         23 . The method of  claim 16 , wherein the method further comprises making a nucleic acid sequence encoding a human V L  domain that is cognate with the human V H  domain, comprising isolating a B cell encoding the human V H  domain and the human V L  domain from said mouse and obtaining therefrom the sequence of the heavy and light chain variable domains. 
     
     
         24 . The method of  claim 23 , wherein the human V L  domain is a V κ  domain. 
     
     
         25 . The method of  claim 16 , wherein the mouse comprises a deletion of the endogenous immunoglobulin heavy chain locus and the endogenous κ light chain. 
     
     
         26 . A rat or mouse comprising an endogenous immunoglobulin heavy chain locus, wherein the endogenous immunoglobulin heavy chain locus consists essentially of
 a) one or more polymorphic variants of a single human V H  gene segment,   b) one or more human D H  gene segments, and   c) one or more human J H  gene segments,   wherein the single human V H  gene segment is a polymorphic V H  gene segment associated with a high copy number in human populations, and   wherein the single human V H  gene segment, one or more human D H  gene segments, and one or more J H  gene segments are operably linked to a non-human immunoglobulin heavy chain constant region gene.   
     
     
         27 . A cell or tissue derived from the rat or mouse of  claim 26 . 
     
     
         28 . A method of making a nucleic acid sequence encoding a human V H  domain, comprising
 (a) immunizing a rat or mouse of  claim 26  with an antigen of interest;   (b) allowing said mouse to mount an immune response to the antigen of interest; and,   (c) obtaining a nucleic acid sequence encoding a human V H  domain from said mouse.