B-NDG B2m plus/hIL15 mice

Basic Information

Description

The highly immunodeficient B-NDG B2m plus/hIL15 mice (NOD.CB17-Prkdc^scidIl2rg^tm1B2m^tm1Fcgrt^{tm1(B2m/Fcgrt)}Il15^tm1(IL15)/Bcgen) was independently designed and generated by Biocytogen and is suitable for engraftment and growth of human tumor cells or tissues.

Targeting strategy

Gene targeting strategy for B-NDG B2m plus/hIL15 mice.

The murine B2m gene was knocked out, and a fused gene composed of murine B2m and Fcgrt was inserted after the signal peptide sequence of the murine Fcgrt gene to generate B-NDG B2m plus/hIL15 mice.

To express human IL15, the CDS region of human IL15 gene was inserted after the 5′UTR of the mouse Il15 gene in B-NDG B2m plus/hIL15 mice.

Protein expression analysis

Strain specific MHC I molecule H-2K^d expression analysis in homozygous B-NDG B2m plus/hIL15 mice by flow cytometry. Splenocytes and bone marrow cells were collected from B-NDG mice and homozygous B-NDG B2m plus/hIL15 mice, and analyzed by flow cytometry with species-specific anti-H-2K^d antibody. Mouse H-2K^d was detectable in B-NDG mice but not in homozygous B-NDG B2m plus/hIL15 mice. The result demonstrates that due to B2m gene knockout, mouse MHC I is not expressed in B-NDG B2m plus/hIL15 mice.

Strain specific IL15 expression analysis in homozygous B-NDG B2m plus/hIL15 mice by ELISA. Serum was collected from B-NDG mice and homozygous B-NDG B2m plus/hIL15 mice which were stimulated with poly(I:C) in vivo, and analyzed by ELISA with species-specific anti-hIL15 antibody. Human IL15 was detectable in homozygous B-NDG B2m plus/hIL15 mice but not in B-NDG mice.

Human PBMC Engraftment

 

Engraftment of human PBMC in B-NDG B2m plus/hIL15 mice alleviated GvHD and enhanced reconstitution of human NK cells. Female B-NDG mice and B-NDG B2m plus/hIL15 mice were respectively engrafted intravenously with human PBMCs on day 0 (n=6).  Peripheral blood was collected to analyze the reconstitution level of human immune cells by FACS. A. Survival rates of the mice were analyzed with Kaplan Meier survival curves. B. Body weight changes. C. Cell number analysis of hCD45+ cells, hCD3+ cells and hCD56+ cells by FACS. D. Representative FACS plots. Results showed that B-NDG B2m plus/hIL15 mice have extended life span and reduced GvHD with human PBMC engraftment when compared to B-NDG mice. Human CD3+ cells and CD56+ cells can be reconstituted successfully in B-NDG B2m plus/hIL15 mice. B-NDG B2m plus/hIL15 mice are a suitable mouse model for human T cell and NK cell reconstitution with PBMC engraftment. Values are expressed as mean ± SEM. (Data from innomodels biotechnologies (Beijing) Co., Ltd.)

Posters

SITC 2023: A novel B2M-deficient immunodeficient model expressing human IL-15 for preclinical evaluation of T cell and NK cell-based therapies

Related products

Product name	Product No.
B-NDG B2m KO mice plus	110601
B-NDG hIL15 mice	110600

Summary

Protein expression analysis:

• Mouse MHC I molecule was not expressed in B-NDG B2m plus/hIL15 mice.
• Human IL15 was detectable in homozygous B-NDG B2m plus/hIL15 mice but not in B-NDG mice.

hPBMC engraftment for human immune system reconstitution：

• B-NDG B2m plus/hIL15 mice have extended life span reduced GvHD upon human PBMC engraftment, when compared to B-NDG mice.
• Human CD3+ cells and CD56+ cells can successfully be reconstituted in B-NDG B2m plus/hIL15 mice.