Protein expression analysis
Strain specific IL2RB expression analysis in homozygous B-hIL2RB/hIL15 mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL15 mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB was detectable in T cells and NK cells of homozygous B-hIL2RB/hIL15 mice.
Strain specific IL2RB expression analysis in homozygous B-hIL2RB/hIL15 mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL15 mice that treated with anti-mCD3ε in vivo, and analyzed by flow cytometry with species-specific antibody. Human IL2RB was detectable in CD4+ T cells and CD8+ T cells of homozygous B-hIL2RB/hIL15 mice.
Strain specific IL15 expression analysis in homozygous B-hIL2RB/hIL15 mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-hIL15/hIL15RA mice (H/H), and analyzed by ELISA. Mouse IL15/IL15RA complex was only detectable in wild-type mice but not in homozygous B-hIL2RB/hIL15 mice (A). Human IL15 was exclusivley detectable in homozygous B-hIL2RB/hIL15 mice but not in wild-type mice (B).
Analysis of spleen leukocytes subpopulation
Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). Flow cytometry analysis of the splenocytes were performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, CD4+ T cells and CD8+ T cells in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of spleen leukocyte subpopulations by FACS. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of monocytes, dendritic cells, granulocytes and macrophages in B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of spleen T leukocytes subpopulation
Analysis of T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). The proportion of T cells subpopulation was tested by flow cytometry. A. Representative FACS plots. Single live cells were gated for the CD3+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of blood leukocytes subpopulation
Analysis of blood leukocyte subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). Flow cytometry analysis of the blood cells were performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, CD4+ T cells and CD8+ T cells in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of blood leukocyte subpopulations by FACS. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of monocytes, dendritic cells, granulocyte and macrophages in B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of blood T leukocytes subpopulation
Analysis of T cell subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). The proportion of T cells subpopulation was tested by flow cytometry. A. Representative FACS plots. Single live cells were gated for the CD3+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of lymph node leukocytes subpopulation
Analysis of lymph node leukocyte subpopulations by FACS. Lymph node cells were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). Flow cytometry analysis of the lymph node cells were performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, CD4+ T cells and CD8+ T cells in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.
Analysis of lymph node T leukocytes subpopulation
Analysis of T cell subpopulations by FACS. Lymph node cells were isolated from female C57BL/6 and B-hIL2RB/hIL15 mice (n=3, 7-week-old). The proportion of T cells subpopulation was tested by flow cytometry. A. Representative FACS plots. Single live cells were gated for the CD3+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hIL2RB/hIL15 mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SEM.Summary
Protein expression analysis:
Human IL2RB was detectable in T cells and NK cells of homozygous B-hIL2RB/hIL15 mice.
Human IL15 was detectable in serum of homozygous B-hIL2RB/hIL15 mice.
Leukocytes cell subpopulation analysis：
The humanization of these genes does not change the overall development, differentiation or distribution of immune cell types in spleen, blood and lymph node.
Targeting strategy
