B-NDG B2m plus/hIL15 mice_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen,drug development,antibody discovery

Strain Name	NOD.CB17-Prkdc^scidIl2rg^tm1B2m^tm1Fcgrt^tm1(B2m)Il15^tm1(IL15)/Bcgen	Common Name	B-NDG B2m 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）

Protein expression analysis

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Strain specific H-2kd expression analysis in homozygous B-NDG B2m plus/hIL15 mice by flow cytometry. Splenocytes and BM cells were collected from wild-type mice (+/+;+/+) and homozygous B-NDG B2m 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 plus/hIL15 mice, which represents B2m is knocked out in B-NDG B2m plus/hIL15 mice.

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

HSC Human Immune System Engraftment

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Human immune cell phenotyping in newborn B-NDG B2m 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 plus/hIL15 mice (male or female, n≥10) after treated with 0.8 Gy irradiation. Blood from B-NDG B2m 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 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.

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Absolute count of Human immune cells in newborn B-NDG B2m 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 plus/hIL15 mice (male or female, n≥10) after treated with 0.8 Gy irradiation. Blood from B-NDG B2m 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 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.

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Survival rate and body weight in B-NDG B2m 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 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 plus/hIL15 mice, and this provides a sufficient experimental time window for the preclinical study.

HSC reconstitution with newborn mice

Comparison of newborn and adult humanization. There are different technological approaches for the engraftment of a functional human immune system in immunodeficient mouse models, such as the injection in facial vein, intracardiac, and intrahepatic. Fewer CD34+HSC cells were required in the injection of newborn mouse in facial vein, which can achieve similar reconstruction efficiency with lower cost, and lots of paper have been published using this approach in the human immune system reconstitution mice.