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 V H gene segment, a plurality of human D H gene segments and a plurality of J H 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 V H gene segment, a plurality of human D H gene segments and a plurality of J H gene segments, at the endogenous immunoglobulin heavy chain locus.

1 . A rat or mouse whose germline genome comprises:
 a restricted immunoglobulin heavy chain locus characterized by the presence of a single human unrearranged V H  gene segment, one or more human unrearranged D H  gene segments, and one or more unrearranged human J H  gene segments.   
     
     
         2 . The rat or mouse of  claim 1 , wherein the rat or mouse comprises a replacement of:
 (i) a plurality of endogenous V H  gene segments,   (ii) all endogenous D H  gene segments, and   (iii) all endogenous J H  gene segments   with   (i) the single human unrearranged V H  gene segment,   (ii) the one or more human unrearranged D H  gene segments, and   (iii) the one or more unrearranged human J H  gene segments.   
     
     
         3 . The rat or mouse of  claim 1 , wherein the single human V H  gene segment is selected from Table 1. 
     
     
         4 .- 5 . (canceled) 
     
     
         6 . The rat or mouse of  claim 1 , wherein the single human unrearranged V H  gene segment, one or more human unrearranged D H  gene segments, and one or more unrearranged human J H  gene segments are operably linked to a human or endogenous immunoglobulin constant region gene. 
     
     
         7 . The rat or mouse of  claim 6 , wherein the immunoglobulin constant region gene is a mouse 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 one or more human J L  gene segments are operably linked to an endogenous light chain constant gene. 
     
     
         11 . The rat or mouse of  claim 10 , wherein the endogenous light chain constant gene is selected from a mouse κ or λ constant region gene. 
     
     
         12 . A mouse whose germline genome comprises a replacement of:
 (i) all endogenous V H  gene segments,   (ii) all endogenous D H  gene segments, and   (iii) all endogenous J H  gene segments   with   (i) a single unrearranged human V H  gene segment and/or polymorphic variants thereof,   (ii) a plurality of unrearranged human D H  gene segments, and   (iii) a plurality of unrearranged human J H  gene segments such that the (i) single unrearranged human gene segment and/or polymorphic variants thereof, (ii) plurality of unrearranged human D H  gene segments, and (iii) plurality of unrearranged human J H  gene segments are operably linked to an endogenous immunoglobulin heavy chain constant region nucleic acid sequence comprising at least an endogenous IgM gene at an endogenous heavy chain locus,   wherein the (i) single unrearranged human gene segment and/or polymorphic variants thereof, (ii) plurality unrearranged human D H  gene segments, and (iii) plurality of unrearranged human J H  gene segments are capable of rearranging and forming a plurality of distinct rearrangements,   wherein each of the plurality of distinct rearrangements is derived from the single human gene segment and/or polymorphic variants thereof, one of the plurality of unrearranged human D H  segments, and one of the all unrearranged human J H  segments, and   wherein each of the plurality of distinct rearrangement encodes a different heavy chain variable domain.   
     
     
         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 . The mouse of  claim 12 , wherein the single human V H  gene segment is selected from Table 1. 
     
     
         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 .- 22 . (canceled) 
     
     
         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 Vκ 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, 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 an endogenous 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 the rat or 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 the rat or mouse.   
     
     
         29 . A nucleic acid construct comprising in operable linkage from 5′ to 3′
 (i) only a single unrearranged human immunoglobulin segment, 
 (ii) one or more unrearranged human immunoglobulin D H  gene segments, 
 (iii) one or more unrearranged human immunoglobulin J H  gene segments, and 
 (iv) at least one homology arm homologous to a rat or mouse immunoglobulin heavy chain locus, 
 wherein upon recombination, the (i) single unrearranged human immunoglobulin segment and/or polymorphic variants thereof, (ii) one or more unrearranged human immunoglobulin D H  gene segments, and (iii) one or more unrearranged human immunoglobulin J H  gene segment are operably linked to at least an endogenous IgM gene at the rate or mouse immunoglobulin heavy chain locus. 
 
     
     
         30 . The nucleic acid construct of  claim 29  comprising in operable linkage from 5′ to 3′
 (i) the single unrearranged human immunoglobulin segment, 
 (ii) a plurality of unrearranged human immunoglobulin D H  gene segments, 
 (iii) a plurality of unrearranged human immunoglobulin J H  gene segments, and 
 (iv) the at least one homology arm homologous to a rat or mouse immunoglobulin heavy chain locus. 
 
     
     
         31 . A rat or mouse comprising the nucleic acid construct of  claim 29 . 
     
     
         32 . A cell or tissue derived from the rat or mouse of  claim 31 . 
     
     
         33 . A method of making a nucleic acid sequence encoding a human V H  domain, comprising
 (a) immunizing a rat or mouse of  claim 31  with an antigen of interest;   (b) allowing the rat or 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 the rat or mouse.