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NZ766708B2 - Transgenic mice expressing chimeric major histocompatibility complex (mhc) class ii molecules - Google Patents
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NZ766708B2 - Transgenic mice expressing chimeric major histocompatibility complex (mhc) class ii molecules - Google Patents

Transgenic mice expressing chimeric major histocompatibility complex (mhc) class ii molecules

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Publication number
NZ766708B2
NZ766708B2 NZ766708A NZ76670812A NZ766708B2 NZ 766708 B2 NZ766708 B2 NZ 766708B2 NZ 766708 A NZ766708 A NZ 766708A NZ 76670812 A NZ76670812 A NZ 76670812A NZ 766708 B2 NZ766708 B2 NZ 766708B2
Authority
NZ
New Zealand
Prior art keywords
mhc
human
polypeptide
rodent
chimeric
Prior art date
Application number
NZ766708A
Other versions
NZ766708A (en
Inventor
Cagan Gurer
Lynn Macdonald
Andrew J Murphy
Sean Stevens
Naxin Tu
Vera Voronina
Original Assignee
Regeneron Pharmaceuticals Inc
Filing date
Publication date
Application filed by Regeneron Pharmaceuticals Inc filed Critical Regeneron Pharmaceuticals Inc
Publication of NZ766708A publication Critical patent/NZ766708A/en
Publication of NZ766708B2 publication Critical patent/NZ766708B2/en

Links

Abstract

Discloses an isolated non-human animal cell comprising at an endogenous Major Histocompatibility Complex II (MHC II) a gene locus a nucleotide sequence encoding a chimeric human/non-human MHC II a polypeptide, wherein a human portion of the chimeric human/non-human MHC II a polypeptide comprises a human MHC II a2 domain. Also disclosed is an isolated non-human animal cell comprising at an endogenous MHC II ß gene locus a nucleotide sequence encoding a chimeric human/non-human MHC II ß polypeptide, wherein a human portion of the chimeric human/non-human MHC II ß polypeptide comprises a human MHC II ß2 domain.

Claims (45)

What is d is:
1. A nucleic acid comprising a sequence encoding a functional chimeric human/nonhuman MHC II ? polypeptide, wherein a human portion of the functional chimeric human/non-human MHC II ? polypeptide comprises ?1 and ?2 domains of a human MHC II ? polypeptide.
2. The nucleic acid of claim 1, further sing non-human regulatory ts operably linked to the sequence encoding the functional ic human/non-human MHC II ? polypeptide.
3. The nucleic acid of claim 1 or 2, n the non-human portion of the functional chimeric human/non-human MHC II ? polypeptide comprises transmembrane and cytoplasmic domains of a rodent MHC II ? polypeptide.
4. The nucleic acid of claim 3, wherein the rodent MHC II a polypeptide is a mouse MHC II a polypeptide.
5. The nucleic acid of claim 4, wherein the human MHC II ? ptide is selected from the group consisting of an MHC II ? polypeptide of HLA-DR, HLA-DQ, and HLA-DP, and the mouse MHC II ? polypeptide is an MHC II ? polypeptide of H-2E.
6. A nucleic acid comprising a sequence encoding a functional chimeric human/nonhuman MHC II ? polypeptide, wherein a human portion of the functional chimeric human/non-human MHC II ß polypeptide ses ?1 and ?2 domains of a human MHC II ? polypeptide.
7. The nucleic acid of claim 6, further comprising man regulatory elements operably linked to the sequence encoding the functional chimeric human/non-human MHC II ? polypeptide.
8. The nucleic acid of claim 6 or 7, wherein the non-human portion of the functional chimeric human/non-human MHC II ß polypeptide comprises transmembrane and cytoplasmic domains of a rodent MHC II ? polypeptide.
9. The nucleic acid of claim 8, n the rodent MHC II ß polypeptide is a mouse MHC II ß polypeptide.
10. The c acid of claim 9, wherein the human MHC II ? polypeptide is selected from the group consisting of an MHC II ? polypeptide of HLA-DR, HLA-DQ, and HLA-DP and the mouse MHC II ? polypeptide is an MHC II ? polypeptide of H-2E.
11. A functional chimeric human/non-human animal MHC II ? polypeptide, wherein a human portion of the functional chimeric human/non-human animal MHC II a polypeptide comprises ?1 and ?2 domains of a human MHC II ? polypeptide and a non-human portion of the functional chimeric human/non-human animal MHC II a polypeptide comprises transmembrane and cytoplasmic s of a man animal MHC II ? polypeptide.
12. The onal chimeric human/non-human animal MHC II ? polypeptide of claim 11, wherein the non-human animal MHC II ? polypeptide is a rodent MHC II ? polypeptide.
13. The functional chimeric human/non-human animal MHC II ? polypeptide of claim 12, wherein the rodent MHC II ? ptide is a mouse MHC II ? polypeptide.
14. The functional chimeric human/non-human animal MHC II ? polypeptide of claim 13, wherein the human MHC II ? polypeptide is selected from the group consisting of an MHC II ? polypeptide of HLA-DR, HLA-DQ, and HLA-DP, and the mouse MHC II ? polypeptide is an MHC II ? polypeptide of H-2E.
15. A functional chimeric human/non-human animal MHC II ? polypeptide, n a human portion of the functional chimeric human/non-human animal MHC II ? polypeptide ses ?1 and ?2 domains of a human MHC II ? polypeptide and a non-human portion of the functional chimeric human/non-human animal MHC II ß ptide comprises transmembrane and cytoplasmic domains of a non-human animal MHC II ? polypeptide.
16. The functional chimeric human/non-human animal MHC II ß polypeptide of claim 15, wherein the non-human animal MHC II ß polypeptide is a rodent MHC II ß ptide.
17. The functional chimeric human/non-human animal MHC II ß ptide of claim 16, wherein the rodent MHC II ß polypeptide is a mouse MHC II ß polypeptide.
18. The functional chimeric human/non-human animal MHC II ß polypeptide of claim 17, n the human MHC II ß polypeptide is ed from the group consisting of an MHC II ß polypeptide of HLA-DR, HLA-DQ, and HLA-DP and the mouse MHC II ß polypeptide is an MHC II ß polypeptide of H-2E.
19. A functional chimeric human/non-human animal MHC II protein complex sing (i) a chimeric human/non-human animal MHC II ? polypeptide, n a human portion of the chimeric human/non-human animal MHC II ? polypeptide comprises ?1 and ?2 domains of a human MHC II ? polypeptide and a non-human portion of the chimeric human/non-human animal MHC II ? polypeptide comprises transmembrane and cytoplasmic domains of a non-human animal MHC II ? polypeptide; and (ii) a chimeric human/non-human animal MHC II ? polypeptide, wherein a human portion of the chimeric human/non-human animal MHC II ? polypeptide comprises ?1 and ?2 domains of a human MHC II ? polypeptide and a non-human portion of the chimeric human/non-human animal MHC II ? polypeptide comprises transmembrane and cytoplasmic domains of a non-human animal MHC II ? polypeptide.
20. The functional chimeric human/non-human animal MHC II protein complex of claim 19, wherein (i) the non-human animal MHC II a polypeptide is a rodent MHC II a polypeptide and (ii) the non-human animal MHC II ß polypeptide is a rodent MHC II ß polypeptide.
21. The functional chimeric human/non-human animal MHC II protein complex of claim 20, n (i) the rodent MHC II a polypeptide is a mouse MHC II a polypeptide and (ii) the rodent MHC II ß ptide is a mouse MHC II ß polypeptide.
22. The functional chimeric non-human animal MHC II protein x of claim 21, wherein the human MHC II ? polypeptide is selected from the group consisting of an MHC II ? polypeptide of HLA-DR, HLA-DQ, and HLA-DP, the human MHC II ? polypeptide is selected from the group consisting of an MHC II ? polypeptide of HLA-DR, HLA-DQ, and HLA-DP, and the mouse MHC II ? and ? polypeptides are respectively MHC II ? and ? polypeptides of H-2E.
23. A method of making a genetically modified rodent comprising inserting, at an endogenous Major Histocompatibility Complex II (MHC II) ? gene locus, a tide molecule encoding a ic human/rodent MHC II ? polypeptide that comprises a human portion and a rodent portion and/or inserting, at an nous MHC II ? gene locus, a nucleotide molecule encoding a chimeric human/rodent MHC II ? polypeptide that comprises a human portion and a rodent portion, wherein the human portion of the chimeric rodent MHC II ? polypeptide comprises a human MHC II ?2 domain, and/or the human portion of the chimeric human/rodent MHC II ? polypeptide comprises a human MHC II ?2 , and n the rodent expresses a functional chimeric human/rodent MHC II complex on a surface of an antigen presenting cell of the rodent.
24. The method of claim 23, wherein the chimeric human/rodent MHC II ? ptide further ses a human ?1 domain.
25. The method of claim 23 or claim 24, wherein the ic human/rodent MHC II ? polypeptide further comprises a human ?1 .
26. The method of any one of claims 23-25, n the tide molecule ng the chimeric human/rodent MHC II ? polypeptide is expressed under regulatory control of endogenous rodent MHC II ? promoter and tory elements and/or the nucleotide molecule encoding the chimeric human/rodent MHC II ? polypeptide is expressed under regulatory control of endogenous rodent MHC II ? er and regulatory elements.
27. The method of any one of claims 23-26, wherein the rodent portion of the chimeric human/rodent MHC II ? polypeptide comprises transmembrane and cytoplasmic domains of an endogenous rodent MHC II ? polypeptide and/or the rodent portion of the chimeric human/rodent MHC II ? polypeptide comprises transmembrane and cytoplasmic domains of an endogenous rodent MHC II ? polypeptide.
28. The method ing to any one of claims 23-27, comprising ing, at the endogenous MHC II ? gene locus, the nucleotide molecule encoding the chimeric human/rodent MHC II ? polypeptide and inserting, at the endogenous MHC II ß gene locus, the nucleotide molecule encoding the chimeric human/rodent MHC II ß polypeptide.
29. The method of claim 28, wherein the rodent does not express, on the surface of its antigen presenting cells, functional endogenous MHC II polypeptides from the endogenous rodent MHC II gene loci.
30. The method of claim 28 or claim 29, n the nucleotide molecule encoding the chimeric human/rodent MHC II ? polypeptide is expressed under regulatory control of endogenous rodent MHC II ? promoter and regulatory elements, and the nucleotide molecule encoding the chimeric human/rodent MHC II ß polypeptide is expressed under regulatory control of endogenous rodent MHC II ß promoter and regulatory elements.
31. The method of any of claims 23-30, wherein the human portion of the chimeric rodent MHC II a polypeptide is encoded by a HLA class II ? chain gene selected from the group ting of an HLA-DR a chain gene, an HLA-DQ ? chain gene, and an HLA-DP ? chain gene and/or the human portion of the chimeric human/rodent MHC II ß polypeptide is encoded by a human HLA class II ß chain gene selected from the group consisting of an HLA-DR ß chain gene, an HLA-DQ ß chain gene, and an HLA-DP ß chain gene.
32. The method of claim 31, wherein the human portion of the chimeric human/rodent MHC II a polypeptide is encoded by a human HLA-DRA gene and/or the human portion of the chimeric human/rodent MHC II ß polypeptide is encoded by a human HLA-DRB gene.
33. The method of claim 32, wherein the human portion of the chimeric human/rodent MHC II ? polypeptide is encoded by a human HLA-DRA gene and/or the human portion of the chimeric human/rodent MHC II ß polypeptide is encoded by a human HLA-DR4 ß gene.
34. The method of claim 32 or claim 33, wherein the human portion of the chimeric human/rodent MHC II ? polypeptide is d by a human HLA-DRA gene and the human portion of the chimeric rodent MHC II ß polypeptide is d by a human HLA- DRB1*04 gene.
35. The method of any one of claims 23-34, wherein the rodent is a rat.
36. The method of any one of claims 23-34, wherein the rodent is a mouse.
37. The method of claim 36, wherein the rodent portions of the chimeric human/rodent MHC II a and/or ß ptides are encoded by mouse H-2E ? and/or mouse H-2E ß chain genes, respectively.
38. The method of claim 36, comprising inserting, at an endogenous MHC II ? gene locus, a nucleotide molecule encoding a chimeric human/mouse MHC II ? polypeptide that comprises a human portion and a mouse portion and inserting, at an endogenous MHC II ß gene locus, a nucleotide le encoding a chimeric human/mouse MHC II ß polypeptide that comprises a human n and a mouse portion, wherein the human portion of the ic human/mouse MHC II ? polypeptide comprises ?1 and ?2 domains encoded by a human HLA-DR4 ? chain gene and the human portion of the chimeric MHC II ß polypeptide comprises ß1 and ß2 domains encoded by a human HLA-DR4 ß chain gene, wherein the mouse portion of the chimeric human/mouse MHC II ? ptide ses transmembrane and cytoplasmic s of a mouse H-2E a chain gene and the mouse portion of the chimeric human/mouse MHC II ß polypeptide comprises transmembrane and cytoplasmic domains of a mouse H-2E ß chain gene, wherein the mouse expresses a functional chimeric human/mouse HLA-DR4/H-2E MHC II complex on a surface of a cell of the mouse, and wherein the functional chimeric human/mouse HLA-DR4/H-2E MHC II complex comprises the chimeric human/mouse MHC II a polypeptide and the chimeric human/mouse MHC II ß polypeptide.
39. The method of any of claims 23-38, wherein inserting, at the endogenous MHC II ? gene locus, comprises replacing a nucleotide molecule encoding a rodent MHC II ?2 domain with a nucleotide molecule encoding a human MHC II ?2 domain and/or inserting, at the endogenous MHC II ß gene locus, comprises replacing a nucleotide molecule encoding a rodent MHC II ß2 domain with a tide molecule ng a human MHC II ß2 domain.
40. The method of claim 39, wherein any replacement is made in a single ES cell, and the single ES cell is introduced into a mouse embryo to make a mouse.
41. The method of claim 39 or claim 40, further comprising replacing, at the endogenous MHC II a gene locus, a nucleotide le encoding a rodent MHC II ?1 domain with a nucleotide molecule encoding a human MHC II ?1 domain replacing, at the endogenous MHC II ß gene locus, a tide molecule encoding a rodent MHC II ß1 domain with a nucleotide molecule ng a human MHC II ß1 domain.
42. A rodent made according to the method of any one of claims 23-41.
43. The rodent of claim 42, wherein the chimeric human/rodent MHC II complex expressed on the surface of the antigen presenting cell comprises the chimeric human/rodent MHC II a polypeptide and the chimeric human/rodent MHC II ß polypeptide, wherein the human portion of the ic human/rodent MHC II a polypeptide comprises a1 and a2 domains of a human MHC II a polypeptide and the rodent portion of the ic human/rodent MHC II a polypeptide comprises transmembrane and cytoplasmic s of a rodent MHC II a polypeptide; and wherein the human portion of the chimeric human/rodent MHC II ß polypeptide comprises ß1 and ß2 domains of a human MHC II ß polypeptide and the rodent portion of the chimeric human/non-human MHC II ß polypeptide comprises transmembrane and asmic domains of a non-human animal MHC II ß ptide.
44. A method for identifying a human CD4+ T-cell epitope comprising ng the rodent of claim 43 to an antigen comprising a CD4+ T-cell epitope, allowing the rodent to mount an immune response, isolating from the rodent an MHC class II-restricted CD4+ T-cell that binds the epitope; and identifying the epitope bound by the MHC class II-restricted CD4+ T-cell.
45. A tissue derived from a rodent made ing to the method of any one of claims 23-41, or the rodent of claim 42 or claim 43, wherein the tissue comprises the antigen presenting cell that ses the functional chimeric human/rodent MHC II complex on the surface of the antigen presenting cell. 2.5 532 m: m a “<33. (V JO—H— .mum-_.w_ a u M. m mcmw <-_ mmmi U\m_._ :98 mammousmma Eot mmmi :98 NSmmYmNnE m. Econuciéoz 35KB o-_ *0 Econocsn. NRMNXO. mare mn m4 nu m . m m m m u u 4 n p mammouzmma mv .0_n_ a a H m.-. m.-. :98 Ecoruc?éoz _wum-_n_ .35.. .89.; Emu-— Emu-— _wum-_n_ Emu-— _wum-_a NNBmYMNn—m Emu-— :50... .08-?— H 95» Econucsu. ”<35. conmmz r995 V mo \_mum-_n_ 5 M... .GE vim a Bum-.."— o.-.“ 3w“: o_<_>_ cobE+mcoxm mm ow d.9“. unatu 35:3: Elm 36 n=<_>_ men HHHm omw? m-— 032 _ cob£+mcoxw me gaziiiwvx thNXE <-_ NhMNxE <-_ cob£+mcoxw +>>\owo? a a m-_ m.-. Siam: cob:_+m:oxw 5 mm m.-. a n m: man-*- a 31 m. .2 <-_ cob£+mcoxa 10/12 E Eng?» mam—u 3a 3:2: 62:5ch swim :5 85w 2 Ha mn— (||.|II.||I|||
NZ766708A 2012-10-26 Transgenic mice expressing chimeric major histocompatibility complex (mhc) class ii molecules NZ766708B2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201161552584P 2011-10-28 2011-10-28
NZ75238412 2012-10-26

Publications (2)

Publication Number Publication Date
NZ766708A NZ766708A (en) 2024-04-26
NZ766708B2 true NZ766708B2 (en) 2024-07-30

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