Strain Name
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C57BL/6-Il2tm1(IL2)Il2ratm1(IL2RA)ll2rbtm2(IL2RB)ll2rgtm2(IL2RG)/Bcgen
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Common Name
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B-hIL2/hIL2R mice
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Background
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C57BL/6
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Catalog number
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112748
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Aliases
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IL2 also known as TCGF, lymphokine
IL2RA also known as CD25, IDDM10, IL2R, IMD41, TCGFR, p55
IL2RB also known as CD122.
IL2RG also known as CD132, SCIDX, IL-2RG
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Protein expression analysis
Strain specific IL2 expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by ELISA. Serum was collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice stimulated with anti-mCD3 and anti-mCD28 in vivo, and analyzed by ELISA with species-specific IL2 ELISA kit. Mouse IL2 was detectable in wild-type mice. Human IL2 was exclusively detectable in homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice.(ND: Not detectable)
Strain specific IL2RA expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice, and analyzed by flow cytometry with the species-specific anti-IL2RA antibody. Human IL2RA was exclusively detectable in T cells, CD4+ T cells and Treg cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice. However, IL2RA was not detectable in CD8+ T cells in both these two mice.
Strain specific IL2RA expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with the species-specific anti-IL2RA antibody. Human IL2RA was exclusively detectable in T cells, CD4+ T cells, CD8+ T cells and Treg cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice.
Strain specific IL2RB expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice, and analyzed by flow cytometry with the species-specific anti-IL2RB antibody. Human IL2RB was exclusively detectable in T cells, CD4+ T cells and CD8+ T cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice.
Strain specific IL2RB expression analysis in homozygous B-hIL2/hIL2RA/hIL2RB mice by flow cytometry. Splenocytes were collected from wild-type mice and homozygous B-hIL2/hIL2RA/hIL2RB mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with the species-specific anti-IL2RB antibody. Human IL2RB was exclusively detectable in T cells, CD4+ T cells and CD8+ T cells of homozygous B-hIL2/hIL2RA/hIL2RB mice but not in wild-type mice. Mouse IL2RA was only detectable in these cells of wild-type mice.
Strain specific IL2RB and IL2RG expression analysis in heterozygous and homozygous B-hIL2RB/hIL2RG mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG mice, and analyzed by flow cytometry with species-specific antibody. Human IL2RB was detectable in heterozygous (H/+;H/+) and homozygous B-hIL2RB/hIL2RG mice (H/H;H/H). Human IL2RG was too low to be detectable in heterozygous, but detectable in homozygous B-hIL2RB/hIL2RG mice.
Strain specific IL2RB and IL2RG expression analysis in heterozygous and homozygous B-hIL2RB/hIL2RG mice by flow cytometry. Splenocytes were collected from wild-type mice and B-hIL2RB/hIL2RG mice, and analyzed by flow cytometry with species-specific antibody. Human IL2RB was detectable in heterozygous (H/+;H/+) and homozygous (H/H;H/H) B-hIL2RB/hIL2RG mice. Human IL2RG was too low to be detectable in heterozygous mice, and it was unable to judge the IL2RG expression in NK cells, because NK cells were diminished in homozygous B-hIL2RB/hIL2RG mice.
Phosphorylation of STAT5 induced by human IL2 and complex of IL15/IL15RA
Intracellular pSTAT5 analysis in spleen T cells of B-hIL2RB/hIL2RG mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice, homozygous and heterozygous B-hIL2RB/hIL2RG mice, and stimulated with IL2 or complex of IL15/IL15RA in vitro for 30 min. Then analyzed by flow cytometry with anti-pSTAT5 antibodies. Human IL2 induced a low phosphorylation level of STAT5 in C57BL/6 mice compared with humanized mice. As for the IL15/IL15RA complex, both mouse and human complex can induce the pSTAT5, but the mouse complex results low in homozygous mice. Above all, the IL2RB and IL2RG humanization did not change the signaling pathway.
Analysis of leukocytes cell subpopulation in spleen
Analysis of spleen leukocyte subpopulations by flow cytometry. Splenocytes were isolated from male C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was 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 and monocytes decreased in humanized mice, while percent of B cells increased in humanized mice. NK cells, dendritic cells, neutrophils and macrophages in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of T cell subpopulation in spleen
Analysis of spleen T cell subpopulations by flow cytometry. Splenocytes were isolated from female C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of leukocytes cell subpopulation in blood
Analysis of blood leukocyte subpopulations by flow cytometry. Blood cells were isolated from male C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the blood cells was 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, neutrophils and monocytes decreased in humanized mice, while percent of B cells increased in humanized mice. NK cells, dendritic cells and macrophages in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of T cell subpopulation in blood
Analysis of blood T cell subpopulations by flow cytometry. Blood cells were isolated from female C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the blood cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of leukocytes cell subpopulation in lymph nodes
Analysis of lymph nodes leukocyte subpopulations by flow cytometry. Lymph nodes cells were isolated from male C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the lymph nodes cells was 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 decreased in humanized mice, while percent of B cells increased in humanized mice. NK cells in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of T cell subpopulation in lymph nodes
Analysis of lymph nodes T cell subpopulations by flow cytometry. Lymph nodes cells were isolated from female C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the lymph nodes cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells decreased in humanized mice, while CD4+ T cells increased in humanized mice. Tregs in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.
Analysis of leukocytes cell subpopulation in thymus
Analysis of thymus leukocyte subpopulations by flow cytometry. Thymus cells were isolated from male C57BL/6 and B-hIL2/hIL2R mice (n=3, 7-week-old). Flow cytometry analysis of the thymus cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and gated for CD3+ T cell population used for further analysis as indicated here. B-C. Results of FACS analysis. Percent of T cells, CD4+ T cells and CD8+ T cells in homozygous B-hIL2/hIL2R mice were similar to those in the C57BL/6 mice. Values are expressed as mean ± SD.