Humanized T cell co-receptor mice
The invention provides genetically modified non-human animals that express chimeric human/non-human T cell co-receptor polypeptides (e.g., CD4, CD8α, CD8β), 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.
1. A genetically modified rodent, comprising
a nucleotide sequence encoding a chimeric human/rodent T cell CD4 co-receptor polypeptide,
wherein a human portion of the human/rodent T cell CD4 co-receptor polypeptide comprises at least D1, D2 and D3 domains of a human CD4 polypeptide,
wherein a rodent portion of the chimeric human/rodent T cell CD4 co-receptor polypeptide comprises at least D4, transmembrane, and cytoplasmic domains of a rodent T cell co-receptor, and
wherein the rodent expresses the chimeric human/rodent T cell CD4 co-receptor polypeptide,
wherein the rodent is a rat or a mouse.
2. The genetically modified rodent of claim 1 , wherein the nucleotide sequence is present at an endogenous rodent T cell CD4 co-receptor locus.
3. The genetically modified rodent of claim 1 , wherein the nucleotide sequence is comprised in the germline of the rodent.
4. The genetically modified rodent of claim 1 , wherein the nucleotide sequence comprises in operable linkage
(a) a sequence encoding an endogenous rodent CD4 signal peptide;
(b) a sequence encoding human CD4 D1-D3 domains; and
(c) a sequence encoding an endogenous rodent D4, transmembrane and cytoplasmic domains.
5. The genetically modified rodent of claim 1 , wherein the rodent does not express a functional endogenous rodent T cell CD4 co-receptor from its endogenous rodent T cell CD4 co-receptor locus.
6. The genetically modified rodent of claim 1 , wherein the chimeric human/rodent T cell CD4 co-receptor polypeptide is not expressed on B cells or CD8 single positive T cells of the rodent.
7. The genetically modified rodent of claim 1 , wherein the human portion of the chimeric human/rodent T cell CD4 co-receptor polypeptide is set forth in SEQ ID NO:57.
8. The genetically modified rodent of claim 1 , further comprising a human or humanized MHC II protein that comprises an extracellular domain of a human MHC II α polypeptide and an extracellular domain of a human MHC IIβ polypeptide.
9. The genetically modified rodent of claim 1 , wherein the rodent is a mouse.
10. The genetically modified rodent of claim 1 , wherein the rodent is a mouse, and wherein the chimeric human/rodent T cell CD4 co-receptor polypeptide is set forth in SEQ ID NO:4.
11. The genetically modified rodent of claim 10 , wherein the mouse further expresses a human or humanized MHC II protein that comprises α1 and α2 domains of a human MHC IIα a polypeptide and β1 and β2 domains of a human MHC II β polypeptide.
12. A method for making the rodent of claim 1 , comprising
replacing at an endogenous CD4 locus of a rodent a nucleotide sequence encoding D1-D3 domains of an endogenous rodent CD4 polypeptide with a nucleotide sequence encoding D1-D3 domains of a human CD4 polypeptide,
wherein the rodent is a rat or a mouse.
13. The method of claim 12 , wherein the endogenous CD4 locus retains an endogenous nucleotide sequence encoding rodent CD4 D4, transmembrane and cytoplasmic domains.
14. The method of claim 12 , wherein the endogenous CD4 locus retains endogenous CD4 promoter and regulatory elements.
15. A chimeric human/rodent CD4 co-receptor polypeptide comprising human D1, D2 and D3 domains and rodent D4, transmembrane and cytoplasmic domains,
wherein the rodent is a rat or a mouse.
16. The chimeric human/rodent CD4 co-receptor polypeptide of claim 15 , wherein the CD4 co-receptor comprises an amino acid sequence set forth in SEQ ID NO:4.
17. A rodent cell comprising a gene encoding the chimeric human/rodent CD4 co-receptor polypeptide, wherein the chimeric human/rodent CD4 co-receptor polypeptide comprises human D1, D2 and D3 domains and rodent D4, transmembrane and cytoplasmic domains, and wherein the cell is an embryonic stem cell, an induced pluripotent cell, a T cell, a hybridoma, or a quadroma,
wherein the rodent cell is a rat cell or a mouse cell.
18. The rodent cell of claim 17 , wherein the CD4 co-receptor polypeptide comprises an amino acid sequence set forth in SEQ ID NO:4.
19. The rodent cell of claim 17 , wherein the rodent cell is a mouse cell.
20. An in vitro composition comprising the rodent cell of claim 17 .
21. A method of using a rodent to generate an HLA class II-restricted immune response comprising:
(i) exposing the rodent of claim 8 to the antigen; and
(ii) detecting the presence of an HLA class II-restricted immune response specific for the antigen in the rodent.
22. The method of claim 21 , further comprising
(iii) identifying the antigen bound by the human or humanized MHC II polypeptide.
23. The method of claim 12 , wherein the rodent is a mouse.
24. The method of claim 23 , wherein the chimeric human/rodent T cell CD4 co-receptor polypeptide comprises an amino acid sequence set forth in SEQ ID NO:4.