B-NDG B2m KO plus/hIL15 mice

Basic Information

Strain name
NOD.CB17-PrkdcscidIl2rgtm1B2mtm1Fcgrttm1(B2m)Il15tm1(IL15)/Bcgen
Common name
B-NDG B2m KO plus/hIL15 mice
Background
B-NDG
Catalog number
111856
Aliases
B2m (beta-2 microglobulin) ; Fcgrt (Fc receptor, IgG, alpha chain transporter,as known as Fcrn) L15  (interleukin 15
NCBI Gene ID

Targeting strategy

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

The exons of B2m were knock out and the CDS region of this gene was fused to FcRn  in B-NDG B2m KO plus/hIL15 mice.

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

Protein expression analysis

Strain specific mH-2kd expression analysis in homozygous B-NDG B2m KO plus/hIL15 mice by flow cytometry. Splenocytes and BM cells were collected from wild-type mice (+/+) and homozygous B-NDG B2m KO plus/hIL15 mice (-/-;H/H),and analyzed by flow cytometry with species-specific anti-H-2kd antibody. Mouse H-2kd was detectable in wild-type mice but not in homozygous B-NDG B2m KO plus/hIL15 mice, which represents B2m is knocked out in B-NDG B2m KO plus/hIL15 mice.

Strain specific IL15 expression analysis in homozygous B-NDG B2m KO plus/hIL15 mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-NDG B2m KO plus/hIL15 mice(-/-;H/H), stimulated with poly(I:C) in vivoand analyzed by ELISA with species-specific IL15 ELISA kit. Human IL15 was detectable in homozygous B-NDG B2m KO plus/hIL15 mice but not in wild-type mice.

HSC Human Immune System Engraftment

Human immune cell phenotyping in newborn B-NDG B2m KO plus/hIL15 mice engrafted with human HSC. PBS or different Human donor CD34+ cells (3E4) were interfacial injected into the newborn homozygote B-NDG B2m KO plus/hIL15 mice (male or female, n≥10) after treated with 0.8 Gy irradiation. Blood from B-NDG B2m KO plus/hIL15 mice was taken at different times for flow cytometric analysis. T cells began to develop at about 10th week and NK cells can maintain at a high level, while the number of NKT cells didn’t change. This suggests that human NK and T cells in reconstituted B-NDG B2m KO plus/hIL15 mice were successfully propagated, and this provides a powerful tool for the preclinical study of antibodies with the function of T cells and NK cells.

Absolute count of Human immune cells in newborn B-NDG B2m KO plus/hIL15 mice engrafted with human HSC. PBS or different Human donor CD34+ cells (3E4) were interfacial injected into the newborn homozygote B-NDG B2m KO plus/hIL15 mice (male or female, n≥10) after treated with 0.8 Gy irradiation. Blood from B-NDG B2m KO plus/hIL15 mice was taken at different times for flow cytometric analysis. T cells began to develop at about 10th week and NK cells can maintain at a high level, while the number of NKT cells didn’t change. This suggests that human NK and T cells in reconstituted B-NDG B2m KO plus/hIL15 mice were successfully propagated, and this provides a powerful tool for the preclinical study of antibodies with the function of T cells and NK cells.

Survival rate and body weight in B-NDG B2m KO plus/hIL15 mice engrafted with human HSC. PBS or different Human donor CD34+ cells (3E4) were interfacial injected into the newborn homozygote B-NDG B2m KO plus/hIL15 mice (male or female, n≥10) after treated with 0.8 Gy irradiation. (A) The survival rate (B) The mean ± SEM of Body weight. This suggests a low GvHD reaction in reconstituted B-NDG B2m KO plus/hIL15 mice, and this provides a sufficient experimental time window for the preclinical study.

Summary

Protein expression analysis:

Mouse H-2kd was detectable in wild-type mice but not in homozygous B-NDG B2m KO plus/hIL15 mice, which represents B2m is knocked out in B-NDG B2m KO plus/hIL15 mice.

Mouse FcRn was both detectable in wild type mice and homozygous B-NDG B2m KO plus/hIL15 mice.

Human IL15 was detectable in homozygous B-NDG B2m KO plus/hIL15 mice but not in wild-type mice.

HSC Human Immune System Engraftment

human NK and T cells in reconstituted B-NDG B2m KO plus/hIL15 mice were successfully propagated with low GvHD reaction.