Basic Information
Description
Biocytogen independently established the highly immunodeficient B-NDG mouse model expressing the human IL15 protein (B: Biocytogen; N: NOD background; D: DANPK (Prkdc) null; G: Il2rg knockout).
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Targeting strategy
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Details
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B-NDG hIL15 Mice are Well Suited for Human Immune System Engraftment
CD34+ constitution standard：hCD45%≥25%
Representative flow cytometric analysis of PBMCs from mice after engraftment with human CD34+ cells. B-NDG hIL15 mice show a significantly higher percentage of human CD45+ cells and of multi-lineage cells,including CD3+ T cells, CD19+ B cells and NK cells. Our results suggest that human B, NK and T cells in reconstituted B-NDG mice were successfully propagated.
Analysis of functions of NK cells in vivo-expanded in B-NDG hIL15 mice after human immune system engraftment.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice (6 week-old, n=5). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice after engraftment with human CD34+ cells. B-NDG hIL15 show a higher percentage of human CD45+ compared with B-NDG. Our results suggest that human leukocytes in reconstituted B-NDG IL15 mice were successfully propagated.
Analysis of functions of NK cells in vivo-expanded in B-NDG hIL15 mice after human immune system engraftment.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice (6 week-old, n=5). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice at 6 week (A) and 8 week (B) after engraftment with human CD34+ cells. B-NDG hIL15 show a higher percentage of human NK cells and the human NK cells express functional proteins.
Analysis of functions of NK cells in vivo-expanded in B-NDG hIL15 mice after human immune system engraftment.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice (6 week-old, n=5). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice at 6 week after engraftment with human CD34+ cells. B-NDG hIL15 show a higher percentage of human NK cells and the human NK cells express functional proteins.
Analysis of functions of NK cells in vivo-expanded in B-NDG hIL15 mice after human immune system engraftment.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice (6 week-old, n=5). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice at 8 week after engraftment with human CD34+ cells. B-NDG hIL15 show a higher percentage of human NK cells and the human NK cells express functional proteins.
Human HSC immune system engraftment and CDX model generation in B-NDG hIL15 mice
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 (female, 7 week-old, n=30) and B-NDG mice (female, 7 week-old, n=30). All mice were treated with 1.6gry-irradiation. Mice were grouped as hCD45% ≥ 10%. K562 cells(1E6), MV-4-11(1E7), Panc-1(5E6) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice respectively. (A-C) The mean ± SEM of tumor sizes. . K562 cells, MV-4-11, Panc-1 were successfully tumorigenic on a mouse model of human HSC immune system reconstitution, and tumor growth was delayed in B-NDG hIL15 mice compared to B-NDG mice.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 (female, 7 week-old, n=30) and B-NDG mice (female, 7 week-old, n=30). All mice were treated with 1.6gry-irradiation. Mice were grouped as hCD45% ≥ 10%. K562 cells(1E6), MV-4-11(1E7), Panc-1(5E6) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice respectively. During the establishment of CDX model using K562 cells (A), MV-4-11 (B), and Panc-1 (C) cells, the proportion of immune cells in the blood of mice were measured. The results showed that during the establishment of tumor models, the proportion of leukocytes, T cells and NK cells in the blood of B-NDG hIL15 mice was higher than B-NDG mice.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice. All mice were treated with 1.6gry-irradiation. Mice were grouped as hCD45% ≥ 10%. MV-4-11(1E7) and Panc-1(5E6) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice respectively. The proportion and function of NK cells in mouse blood and tumor tissues were measured at the end of the experiment. The results showed that B-NDG hIL15 show a higher percentage of human NK cells compared with B-NDG, and NK cell express functional proteins.
Human HSC immune system engraftment and Raji-luc CDX model generation in B-NDG hIL15 mice
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice. All mice were treated with 1.6gry-irradiation. Mice were grouped as hCD45% ≥ 10%. Raji-luc(5E5) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice respectively. (A) fluorescence imager was used to monitor tumor fluorescence in mice. (B-C) Body weight and survival percentage changes during treatment. The results showed that Raji-luc were successfully tumorigenic on a mouse model of human HSC immune system reconstitution, and tumor growth was delayed in B-NDG hIL15 mice compared to B-NDG mice.
Human CD20 antibody in vivo efficacy study using B-NDG hIL15 mice with human HSC immune system reconstitution model.
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL15 and B-NDG mice. All mice were treated with 1.6gry-irradiation. Raji-luc(5E5) were implanted into B-NDG IL15 mice. Mice were grouped when the tumor volume reached approximately 100-150mm3 (n=5) at which time they were treated with rituximab (in house) with doses and schedules indicated in panel . The proportion of leukocyte and NK cells in mouse blood were measured at the different time of the experiment. The results showed that CD20 antibodies had efficacies for tumor growth inhibition in B-NDG hIL15 mice with human HSC immune system reconstitution model.
PBMC Human Immune System Engraftment
Human immune cell phenotyping in B-NDG hIL15 engrafted with human PBMC
Human immune cell phenotyping in B-NDG hIL15 engrafted with human PBMC. Human PBMC cells (5E6) were intravenous implanted into homozygote B-NDG hIL15 (female, 5-week-old, n=15) and B-NDG mice (female, 5-week-old, n=10). Blood from B-NDG hIL15 and B-NDG mice was taken at different times after human PBMC engraftment for flow cytometric analysis. A. Human immune cell phenotyping. B. NK cell phenotyping. C. Survival. D. Body weight. Values were expressed mean ± SEM.
Human PBMC immune system engraftment and K562 CDX model generation in B-NDG hIL15 mice
Analysis of CD45+ and NK cells after PBMC engraftment and K562 CDX model generation in B-NDG hIL15 mice . Human PBMC cells(5E6) were intravenous implanted into homozygote B-NDG hIL15 (female, 6 week-old, n=5) and B-NDG mice (female, 6 week-old, n=5). K562 cells(1E6) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice on the same day as PBMCs. (A) The mean ± SEM of tumor sizes (B) The mean ± SEM of Body weight. (C) Blood from B-NDG hIL15 and B-NDG mice was taken at different times for flow cytometric analysis. K562 cells were successfully tumorigenic on a mouse model of human PBMC immune system reconstitution, and B-NDG hIL15 showed a higher percentage of human CD45+ cells and cell number of human NK cells compared to B-NDG.
Human PBMC immune system engraftment and PANC-1 CDX model generation in B-NDG hIL15 mice
Human PBMC cells(5E6) were intravenous implanted into homozygote B-NDG hIL15 (female, 6 week-old, n=5) and B-NDG mice (female, 6 week-old, n=5). PANC-1 cells(5E6) were subcutaneously implanted into B-NDG hIL15 and B-NDG mice on the same day as PBMCs. (A) The mean ± SEM of tumor sizes (B) The mean ± SEM of Body weight. (C)Blood from B-NDG hIL15 and B-NDG mice was taken at different times for flow cytometric analysis. PANC-1 cells were successfully tumorigenic on a mouse model of human PBMC immune system reconstitution, and tumor growth was significantly delayed in B-NDG hIL15 mice compared to B-NDG mice. B-NDG hIL15 showed a higher percentage of human CD45+ cells and cell number of human NK cells compared to B-NDG.
Antitumor effect of CLDN18.2 antibody in B-NDG hIL15 mice reconstituted with human PBMCs
Human PBMC cells(1E6) were intravenous implanted into homozygote B-NDG hIL15 mice. NUGC4-CLDN18.2(6E6) were implanted into B-NDG IL15 mice. Mice were grouped when the tumor volume reached approximately 70-100mm3 (n=5) at which time they were treated with IMAB362 (in house) with doses and schedules indicated in panel. . (A) Tumor volume changes during treatment, (B) Body weight changes during treatment. The results showed that CLDN18.2 antibodies had efficacies for tumor growth inhibition in B-NDG hIL15 mice with human PBMC immune system reconstitution model.
Antitumor effect of TIGIT antibody in B-NDG hIL15 mice reconstituted with human PBMCs
Human PBMC cells(1E6) were intravenous implanted into homozygote B-NDG hIL15 mice. A375(4E6) were implanted into B-NDG hIL15 mice. Mice were grouped the day after tumor inoculation (n=5) and treated with Tiragolumab (in house) with doses and schedules indicated in panel. (A) Tumor volume changes during treatment, (B) Body weight changes during treatment. The results showed that TIGIT antibodies had efficacies for tumor growth inhibition in B-NDG hIL15 mice with human PBMC immune system reconstitution model.
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References
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1.Ali, A. K., Nandagopal, N. & Lee, S. H. IL-15-PI3K-AKT-mTOR: A Critical Pathway in the Life Journey of Natural Killer Cells. Frontiers in immunology 6, 355, doi:10.3389/fimmu.2015.00355 (2015).
2.Fehniger, T. A. et al. Fatal leukemia in interleukin 15 transgenic mice follows early expansions in natural killer and memory phenotype CD8+ T cells. The Journal of experimental medicine 193, 219-231, doi:10.1084/jem.193.2.219 (2001).
3.Kennedy, M. K. et al. Reversible defects in natural killer and memory CD8 T cell lineages in interleukin 15-deficient mice. The Journal of experimental medicine 191, 771-780, doi:10.1084/jem.191.5.771 (2000).
4.Matsuda, M. et al. Human NK cell development in hIL-7 and hIL-15 knockin NOD/SCID/IL2rgKO mice. Life science alliance 2, doi:10.26508/lsa.201800195 (2019).
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Poster
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B-NDG hIL-15 Mice Improve NK Cell Development After Human PBMC or CD34+ HSC Engraftment
