Genetically modified major histocompatibility complex animals
The invention provides genetically modified non-human animals that express a humanized MHC II protein (humanized MHC II α and β polypeptides), as well as embryos, cells, and tissues comprising the humanized MHC II protein. Also provided are constructs for and methods of making the genetically modified non-human animals. Methods of using the genetically modified non-human animals to study various aspects of the human immune system are provided.
1. A mouse comprising at an endogenous mouse Major Histocompatibility Complex (MHC) II locus a first nucleotide sequence encoding a chimeric human/mouse MHC II α polypeptide and a second nucleotide sequence encoding a chimeric human/mouse MHC II β polypeptide,
wherein a human portion of the chimeric MHC II α polypeptide is encoded by at least a portion of an α chain gene of Human Leukocyte Antigen (HLA)-DR2 and comprises an α2 domain of a HLA-DR2 α polypeptide,
wherein a human portion of the chimeric MHC II β polypeptide is encoded by at least a portion of a β chain gene of HLA-DR2 and comprises a β2 domain of a HLA-DR2 β polypeptide, wherein the first nucleotide sequence is expressed under regulatory control of an MHC II α promoter, and the second nucleotide sequence is expressed under regulatory control of an MHC II β promoter, and
wherein the mouse expresses the chimeric human/mouse MHC II α and β polypeptides on the surface of an antigen presenting cell of the mouse.
2. The mouse of claim 1 , wherein the endogenous mouse MHC II locus is a mouse H-2E locus.
3. The mouse of claim 2 , wherein the endogenous H-2E locus lacks an endogenous nucleotide sequence encoding α1 and α2 domains of a mouse H-2E α polypeptide.
4. The mouse of claim 3 , wherein the endogenous H-2E locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2E β polypeptide.
5. The mouse of claim 2 , wherein the endogenous H-2E locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2E β polypeptide.
6. The mouse of claim 1 , wherein the human portion of the chimeric human/mouse MHC II α polypeptide further comprises a human α1 domain.
7. The mouse of claim 1 , wherein the human portion of the chimeric human/mouse MHC II β polypeptide further comprises a human β1 domain.
8. The mouse of claim 1 , wherein the first nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II α promoter and regulatory elements, and the second nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II β promoter and regulatory elements.
9. The mouse of claim 1 , wherein the chimeric human/mouse MHC II α polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2E α polypeptide.
10. The mouse of claim 1 , wherein the chimeric human/mouse MHC II β polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2E β polypeptide.
11. The mouse of claim 1 , wherein the mouse does not express a functional extracellular domain of a mouse H-2E α polypeptide from an endogenous H-2E locus.
12. The mouse of claim 11 , wherein the mouse does not express a functional extracellular domain of a mouse H-2E β polypeptide from an endogenous H-2E locus.
13. The mouse of claim 1 , wherein the mouse does not express a functional extracellular domain of a mouse H-2E β polypeptide from an endogenous H-2E locus.
14. The mouse of claim 1 , wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2A locus, and wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2E locus.
15. A mouse comprising at an endogenous mouse MHC II locus a first nucleotide sequence encoding a chimeric human/mouse MHC II α polypeptide and a second nucleotide sequence encoding a chimeric human/mouse MHC II β polypeptide, wherein a human portion of the chimeric MHC II α polypeptide is encoded by at least a portion of an α chain gene of HLA-DQ2 and comprises an α2 domain of a HLA-DQ2 α polypeptide, wherein a human portion of the chimeric MHC II β polypeptide is encoded by at least a portion of a β chain gene of HLA-DQ2 and comprises a β2 domain of a HLA-DQ2 β polypeptide, wherein the first nucleotide sequence is expressed under regulatory control of an MHC II α promoter, and the second nucleotide sequence is expressed under regulatory control of an MHC II β promoter, and
wherein the mouse expresses the chimeric human/mouse MHC II α and β polypeptides on the surface of an antigen presenting cell of the mouse.
16. The mouse of claim 15 , wherein the endogenous mouse MHC II locus is an H-2A locus.
17. The mouse of claim 16 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding α1 and α2 domains of a mouse H-2A α polypeptide.
18. The mouse of claim 17 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2A β polypeptide.
19. The mouse of claim 16 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2A β polypeptide.
20. The mouse of claim 15 , wherein the human portion of the chimeric human/mouse MHC II α polypeptide further comprises a human α1 domain.
21. The mouse of claim 15 , wherein the human portion of the chimeric human/mouse MHC II β polypeptide further comprises a human β1 domain.
22. The mouse of claim 15 , wherein the first nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II α promoter and regulatory elements, and the second nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II β promoter and regulatory elements.
23. The mouse of claim 15 , wherein the chimeric human/mouse MHC II α polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2A α polypeptide.
24. The mouse of claim 15 , wherein the chimeric human/mouse MHC II β polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2A β polypeptide.
25. The mouse of claim 15 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A α polypeptide from an endogenous H-2A locus.
26. The mouse of claim 25 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A β polypeptide from an endogenous H-2A locus.
27. The mouse of claim 15 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A β polypeptide from an endogenous H-2A locus.
28. The mouse of claim 15 , wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2A locus, and wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2E locus.
29. A mouse comprising at an endogenous mouse MHC II locus a first nucleotide sequence encoding a chimeric human/mouse MHC II α polypeptide and a second nucleotide sequence encoding a chimeric human/mouse MHC II β polypeptide, wherein a human portion of the chimeric MHC II α polypeptide is encoded by at least a portion of an α chain gene of HLA-DQ8 and comprises an α2 domain of a HLA-DQ8 α polypeptide, wherein a human portion of the chimeric MHC II β polypeptide is encoded by at least a portion of a β chain gene of HLA-DQ8 and comprises a β2 domain of a HLA-DQ8 β polypeptide, wherein the first nucleotide sequence is expressed under regulatory control of an MHC II α promoter, and the second nucleotide sequence is expressed under regulatory control of an MHC II β promoter, and
wherein the mouse expresses the chimeric human/mouse MHC II α and β polypeptides on the surface of an antigen presenting cell of the mouse.
30. The mouse of claim 29 , wherein the endogenous mouse MHC II locus is an H-2A locus.
31. The mouse of claim 30 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding α1 and α2 domains of a mouse H-2A α polypeptide.
32. The mouse of claim 31 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2A β polypeptide.
33. The mouse of claim 30 , wherein the endogenous H-2A locus lacks an endogenous nucleotide sequence encoding β1 and β2 domains of a mouse H-2A β polypeptide.
34. The mouse of claim 29 , wherein the human portion of the chimeric human/mouse MHC II α polypeptide further comprises a human α1 domain.
35. The mouse of claim 29 , wherein the human portion of the chimeric human/mouse MHC II β polypeptide further comprises a human β1 domain.
36. The mouse of claim 29 , wherein the first nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II α promoter and regulatory elements, and the second nucleotide sequence is expressed under regulatory control of endogenous mouse MHC II β promoter and regulatory elements.
37. The mouse of claim 29 , wherein the chimeric human/mouse MHC II α polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2A α polypeptide.
38. The mouse of claim 29 , wherein the chimeric human/mouse MHC II β polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2A β polypeptide.
39. The mouse of claim 29 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A α polypeptide from an endogenous H-2A locus.
40. The mouse of claim 39 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A β polypeptide from an endogenous H-2A locus.
41. The mouse of claim 29 , wherein the mouse does not express a functional extracellular domain of a mouse H-2A β polypeptide from an endogenous H-2A locus.
42. The mouse of claim 29 , wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2A locus, and wherein the mouse does not express a functional endogenous α polypeptide, a functional endogenous β polypeptide, or both from an endogenous H-2E locus.