Humanized mouse model with human immune system

Patent metadata

Field	Value
Publication number	US-2022386573-A1
Application number	US-202017776856-A
Country	US
Kind code	A1
Filing date	Nov 13, 2020
Priority date	Nov 15, 2019
Publication date	Dec 8, 2022
Grant date	—

Abstract

Described herein are transgenic mice for testing immunogenicity and protective efficacy of a wide variety of therapeutic agents and vaccines, determining allograft rejection, and developing monoclonal antibodies and generating hybridomas. Methods of generating a transgenic mouse is also described. Described herein are mouse models capable of expressing B cell, a T cell, a monocyte, a macrophage, a dendritic cell, a NK cell, a iNKT cell, an innate lymphoid cell, a microglia cell, a red blood cell, which can develop into a functional human immune system.

First claim

What is claimed is:

1 . A transgenic mouse, comprising: one or more human CD45 expressing cells, human B cells, human T cells, human dendritic cells, human monocytes/macrophages, human NK cells, human innate lymphoid cells, human microglia or human iNKT cells; and wherein the mouse's endogenous immune system is immunodeficient.

2 . The transgenic mouse of claim 1 , wherein the transgenic mouse comprises one or more mutations, wherein the one or more mutations is:
a) a loss of function mutation causing the mode of action (moa) loss-of-function mutation in the gene that encodes for the protein kinase, DNA-activated, catalytic polypeptide; b) a loss-of-function mutation in the gene that encodes for the interleukin 2 receptor α; or c) a loss-of-function mutation in a gene that encodes for a KIT receptor.

3 . The transgenic mouse of any of claims 1 - 2 , wherein the mouse further comprises an engraftment of human hematopoietic stem cells.

4 . A transgenic mouse, comprising:
a) a loss-of-function mutation in the gene that encodes for the protein kinase, DNA-activated, catalytic polypeptide; and b) a loss-of-function mutation in the gene that encodes for the interleukin 2 receptor α; wherein the mouse further comprises an engraftment of human hematopoietic stem cells, and wherein the mouse comprises one or more human CD45 expressing cells, human B cells, human T cells, human dendritic cells, human monocytes/macrophages, human NK cells, human innate lymphoid cells, human microglia and human iNKT cells.

5 . The transgenic mouse of claim 4 , wherein the mouse does not express a functional DNA-activated, catalytic polypeptide.

6 . The transgenic mouse of claim any of claims 1 - 5 , wherein the engraftment of human hematopoietic stem cells is through intracardial injection.

7 . The transgenic mouse of claim 6 , wherein the mouse does not express a functional interleukin 2 receptor α.

8 . The transgenic mouse of claim 7 , wherein the mouse comprises NOD.Cg-Kit W-41J Prkdc scid Il2rg tm1Wjl WaskJ (NSGW41), NOD.Cg-Kit W-41J Tyr + Prkdc scid Il2rg tm1Wjl /Thom0J (NBSGW) or NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NSG).

9 . The transgenic mouse of any of claims 1 - 8 , wherein the human hematopoietic stem cells comprise one or more cells selected from the group consisting of a human CD34-positive cell, a human hematopoietic stem cell, a human myeloid precursor cell, a human erythroid precursor cell, a human myeloid cell, a human dendritic cell, a human monocyte, a human granulocyte, a human erythrocyte, a human neutrophil, a human mast cell, a human thymocyte, and a human B lymphocyte.

10 . The transgenic mouse of any of claims 1 - 9 , wherein the hematopoietic stem cells are CD34+ stem cells.

11 . The transgenic mouse of any of claims 1 - 10 , wherein the mouse is treated with estrogen or estradiol.

12 . The transgenic mouse of any of claims 1 - 11 , wherein the mouse comprises two or more human CD45 expressing cells, human B cells, human T cells, human dendritic cells, human monocytes/macrophages, human NK cells, human innate lymphoid cells, human microglia and human iNKT cells.

13 . The transgenic of claim 12 , further comprising a loss-of-function mutation in a gene that encodes for a KIT receptor.

14 . The transgenic mouse of claim 13 , wherein the mouse does not express a functional KIT receptor.

15 . The transgenic mouse of any of claims 1 - 14 , wherein the mouse has a functional human immune system.

16 . The transgenic mouse of any of claims 1 - 15 , wherein the loss-of-function mutation in the gene that encodes for the protein kinase, DNA-activated, catalytic polypeptide comprises a T to A transversion point mutation at a position corresponding to codon 4046.

17 . The transgenic mouse of any of claims 1 - 16 , wherein the loss-of-function mutation in the gene that encodes for the protein kinase, DNA-activated, catalytic polypeptide is Prkdc scid .

18 . The transgenic mouse of any of claims 1 - 17 , wherein the loss-of-function mutation in the gene that encodes for the interleukin 2 receptor α comprises a neomycin resistance cassette.

19 . The transgenic mouse of any of claims 1 - 18 , wherein the loss-of-function mutation in the gene that encodes for the interleukin 2 receptor α is Il2rg tm1Wjl .

20 . The transgenic mouse of any of claims 13 - 19 wherein the loss-of-function mutation in the gene that encodes for the KIT receptor comprises a G to A point mutation in a kinase domain at nucleotide 2519.

21 . The transgenic mouse of any of claims 13 - 20 , wherein the loss-of-function mutation in the gene that encodes for the KIT receptor is Cg-Kit W-41J .

22 . The transgenic mouse of any of claims 1 - 21 , wherein the mouse comprises NOD.Cg-Kit W-41J Prkdc scid Il2rg tm1Wjl /WaskJ (NSGW41), NOD.Cg-Kit W-41J Tyr + Prkdc scid Il2rg tm1Wjl /Thom0J (NBSGW) or NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NSG).

23 . The transgenic mouse of any of claims 1 - 22 , wherein the mouse comprises mature human leukocytes.

24 . The transgenic mouse of any of claims 1 - 23 , wherein the mouse comprises one or more human hematopoietic lineage cells.

25 . The transgenic mouse of claim 24 , wherein the one or more human hematopoietic lineage cells is a B cell, a T cell, a monocyte, a macrophage, a dendritic cell, a NK cell, a iNKT cell, an innate lymphoid cell, a microglia or a red blood cell.

26 . The transgenic mouse of claim 25 , wherein the mouse comprises all human hematopoietic lineage cells.

27 . The transgenic mouse of any of claims 25 - 26 , wherein the one or more human hematopoietic lineage cells are maintained up to 40 weeks.

28 . The transgenic mouse of any of claims 1 - 27 , wherein the mouse comprises 95% human cell peripheral reconstitution.

29 . The transgenic mouse of any of claims 1 - 28 , wherein the mouse is capable of supporting full human leukocyte development, differentiation and persistence beyond 1 year of age without developing xeno-reactive graft-versus-host reaction and disease.

30 . The transgenic mouse of any of claims 1 - 27 , wherein the engrafted hematopoietic stem cells are capable of developing into one or more of a human B cell, a human T cell, a human monocyte, a human macrophage, a human dendritic cell, a human NK cell, a human iNKT cell, a human innate lymphoid cell, a human microglia and a human red blood cell or a combination thereof.

31 . The transgenic mouse of of claim 30 , wherein the mouse comprises at least one of each of a human B cell, a human T cell, a human monocyte, a human macrophage, a human dendritic cell, a human NK cell, a human iNKT cell, a human innate lymphoid cell, a human microglia and a human red blood cell.

32 . The transgenic mouse of claim 31 , wherein the at least one of each of a human B cell, a human T cell, a human monocyte, a human macrophage, a human dendritic cell, a human NK cell, a human iNKT cell, a human innate lymphoid cell, a human microglia and a human red blood cell reaches 95% human reconstitution within bone marrow or 98% within a secondary lymphoid organ in response to estrogen stimulation.

33 . The transgenic mouse of claim 32 , wherein the secondary lymphoid organ is a spleen, a mesenteric lymph node or a gut-associated lymphoid tissue.

Assignees

• Univ Texas

Inventors

• Chupp Daniel
• Zan Hong
• Casali Paolo

Classifications

• A01K2227/105

  Murine · CPC title

• A01K67/0271 Primary

  Chimeric vertebrates, e.g. comprising exogenous cells · CPC title

• A01K2267/03

  Animal model, e.g. for test or diseases · CPC title

• A01K2207/12

  Animals modified by administration of exogenous cells · CPC title

• A01K2207/15

  Humanized animals · CPC title