Genetically modified major histocompatibility complex animals

Abstract

The invention provides genetically modified non-human animals that express chimeric human/non-human MHC I polypeptide and/or human or humanized β2 microglobulin polypeptide, as well as embryos, cells, and tissues comprising the same. Also provided are constructs for making said genetically modified animals and methods of making the same. Methods of using the genetically modified animals to study various aspects of human immune system are provided.

What is claimed is: 
     
         1 . A non-human animal comprising at an endogenous Major Histocompatibility Complex I (MHC I) locus a nucleotide sequence encoding a chimeric human/non-human MHC I polypeptide,
 wherein a human portion of the chimeric polypeptide comprises an extracellular domain of a human MHC I polypeptide, and   wherein the non-human animal expresses the chimeric human/non-human MHC I polypeptide.   
     
     
         2 . The animal of  claim 1 , wherein the animal does not express a functional extracellular domain of an endogenous non-human MHC I polypeptide from an endogenous non-human MHC I locus. 
     
     
         3 . The animal of  claim 1 , wherein the nucleotide sequence is operably linked to endogenous non-human regulatory elements. 
     
     
         4 . The animal of  claim 1 , wherein the animal is a rodent. 
     
     
         5 . The rodent of  claim 4 , wherein the rodent is selected from a rat and a mouse. 
     
     
         6 . The rodent of  claim 5 , wherein the rodent is a mouse and the endogenous locus is a mouse H-2D locus. 
     
     
         7 . The animal of  claim 1 , wherein the human portion of the chimeric polypeptide comprises α1, α2, and α3 domains of the human MHC I polypeptide. 
     
     
         8 . The animal of  claim 1 , wherein a non-human portion of the chimeric polypeptide comprises transmembrane and cytoplasmic domains of an endogenous non-human MHC I polypeptide. 
     
     
         9 . The animal of  claim 8 , wherein the animal is a mouse, the endogenous mouse MHC I locus is an H-2D locus, and the endogenous mouse MHC I polypeptide is H-2D. 
     
     
         10 . The animal of  claim 1 , wherein the human MHC I polypeptide is selected from the group consisting of HLA-A, HLA-B, and HLA-C. 
     
     
         11 . The animal of  claim 10 , wherein the human MHC I polypeptide is an HLA-B27 polypeptide. 
     
     
         12 . The animal of  claim 1 , further comprising at its endogenous non-human β2 microglobulin locus a nucleotide sequence encoding a human or humanized β2 microglobulin polypeptide, wherein the animal expresses the human or humanized β2 microglobulin polypeptide. 
     
     
         13 . The animal of  claim 12 , wherein the animal does not express a functional endogenous non-human β2 microglobulin from its endogenous non-human locus. 
     
     
         14 . The animal of  claim 12 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide is operably linked to non-human β2 microglobulin regulatory elements. 
     
     
         15 . The animal of  claim 12 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide comprises a nucleotide sequence set forth in exon 2 to exon 4 of a human β2 microglobulin gene. 
     
     
         16 . The animal of  claim 12 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide comprises a nucleotide sequence set forth in exon 2, exon 3, and exon 4 of a human β2 microglobulin gene. 
     
     
         17 . A mouse comprising at an endogenous H-2D locus a nucleotide sequence encoding a chimeric human/mouse MHC I polypeptide,
 wherein a human portion of the chimeric polypeptide comprises an extracellular domain of a human HLA-B polypeptide and a mouse portion comprises transmembrane and cytoplasmic domains of a mouse H-2D polypeptide, and   wherein the mouse expresses the chimeric HLA-BIH-2D polypeptide.   
     
     
         18 . The mouse of  claim 17 , wherein the mouse does not express a functional extracellular domain of the mouse H-2D polypeptide from an endogenous H-2D locus. 
     
     
         19 . The mouse of  claim 17 , wherein the nucleotide sequence is operably liked to endogenous mouse regulatory elements. 
     
     
         20 . The mouse of  claim 17 , wherein the human portion of the chimeric polypeptide comprises α1, α2, and α3 domains of the human HLA-B polypeptide. 
     
     
         21 . The mouse of  claim 17 , wherein the human HLA-B polypeptide is an HLA-B27 polypeptide. 
     
     
         22 . The mouse of  claim 17 , wherein the nucleotide sequence comprises a sequence of endogenous mouse H-2D leader. 
     
     
         23 . The mouse of  claim 17 , further comprising at its endogenous mouse β2 microglobulin locus a nucleotide sequence encoding a human or humanized β2 microglobulin polypeptide, wherein the animal expresses the human or humanized β2 microglobulin polypeptide. 
     
     
         24 . The animal of  claim 23 , wherein the animal does not express a functional endogenous mouse β2 microglobulin from its endogenous mouse β2 microglobulin locus. 
     
     
         25 . The animal of  claim 23 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide is operably linked to endogenous mouse β2 microglobulin regulatory elements. 
     
     
         26 . The animal of  claim 23 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide comprises a nucleotide sequence set forth in exon 2 to exon 4 of a human β2 microglobulin gene. 
     
     
         27 . The animal of  claim 23 , wherein the nucleotide sequence encoding the human or humanized β2 microglobulin polypeptide comprises a nucleotide sequence set forth in exon 2, exon 3, and exon 4 of a human β2 microglobulin gene. 
     
     
         28 . A non-human animal comprising at an endogenous MHC I locus one, two, three, four, five, or six nucleotide sequence(s) encoding a chimeric human/non-human MHC I polypeptide(s),
 wherein a human portion of the chimeric polypeptide(s) comprises an extracellular domain of a human MHC I polypeptide,   wherein a non-human portion of the chimeric polypeptide(s) comprises transmembrane and cytoplasmic domain of a non-human MHC I polypeptide, and   wherein the animal expresses one, two, three, four, five, or six chimeric human/non-human MHC I polypeptide(s).   
     
     
         29 . The animal of  claim 28 , wherein the animal is a rodent. 
     
     
         30 . The animal of  claim 29 , wherein the rodent is selected from a mouse and a rat. 
     
     
         31 . The rodent of  claim 30 , wherein the rodent is a mouse, and wherein the non-human MHC I polypeptide is a mouse MHC I polypeptide is selected from H-2D, H-2K, and H-2L. 
     
     
         32 . The non-human animal of  claim 28 , wherein the rodent does not express any functional endogenous MHC I polypeptides. 
     
     
         33 . A mouse comprising at an endogenous mouse MHC I locus two nucleotide sequences encoding chimeric human/mouse MHC I polypeptides,
 wherein the two nucleotide sequences encode chimeric HLA-A2/H-2K and HLA-B27/H-2D polypeptides, and   wherein the mouse expresses the chimeric HLA-A2/H-2K and HLA-B27/H-2D polypeptides.   
     
     
         34 . The mouse of  claim 33 , wherein the mouse does not express any functional endogenous mouse MHC I polypeptides. 
     
     
         35 . The mouse of  claim 33 , wherein the nucleotide sequence encoding chimeric HLA-A2/H-2K is located at an endogenous H-2K locus, and the nucleotide sequence encoding chimeric HLA-B27/H-2D is located at an endogenous H-2D locus. 
     
     
         36 . A non-human chimeric MHC I locus encoding a chimeric human/non-human MHC I polypeptide, comprising:
 a first nucleotide sequence encoding a human MHC I extracellular domain operably linked to a second nucleotide sequence encoding a non-human MHC I transmembrane and cytoplasmic domains,   wherein the locus expresses a functional chimeric human/non-human MHC I polypeptide.   
     
     
         37 . The locus of  claim 36 , wherein the MHC I locus is located at an endogenous MHC I position in a genome of a non-human animal. 
     
     
         38 . The locus of  claim 36 , wherein the human MHC I is selected from HLA-A2 and HLA-B27. 
     
     
         39 . The locus of  claim 36 , wherein the non-human MHC I is a mouse MHC I selected from H-2K and H-2D.