Basic Information
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Targeting strategy
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Gene targeting strategy for B-hBDCA2 mice.
The BAC containing the whole human BDCA2 genomic sequences was inserted into mouse Hipp11 gene site in B-hBDCA2 mice.
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Protein expression analysis
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Strain specific BDCA2 expression analysis in B-hBDCA2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and heterozygous B-hBDCA2 mice and homozygous B-hBDCA2 mice, and analyzed by flow cytometry with anti-BDCA2 antibody. Human BDCA2 was exclusively detectable in plasmacytoid dendritic cells (pDCs) of B-hBDCA2 mice but not in wild-type mice. Human BDCA2 was not detectable in T cells, B cells and NK cells of B-hBDCA2 mice and wild-type mice.
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Analysis of leukocytes subpopulation in spleen
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Analysis of spleen leukocyte subpopulations by FACS. Spleen was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-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, B cells, NK cells, dendritic cells, neutrophils, monocytes and macrophages in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that BDCA2 humanized does not change the overall development, differentiation or distribution of these cell types in the spleen. Values are expressed as mean ± SEM.
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Analysis of T cell subpopulation in spleen
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Analysis of spleen T cell subpopulations by FACS. Spleen was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-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 CD3ε+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that BDCA2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the spleen. Values are expressed as mean ± SEM.
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Analysis of leukocytes subpopulations in lymph node
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Analysis of lymph nodes leukocyte subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-week-old). Flow cytometry analysis of the leukocytes 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, B cells, NK cells in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that BDCA2 humanized does not change the overall development, differentiation or distribution of these cell types in the lymph node. Values are expressed as mean ± SEM.
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Analysis of T cell subpopulation in lymph node
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Analysis of lymph nodes T cell subpopulations by FACS. Lymph node was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3ε + T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that BDCA2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the lymph node. Values are expressed as mean ± SEM.
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Analysis of leukocytes subpopulations in blood
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Analysis of blood leukocyte subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood 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, B cells, NK cells, dendritic cells, neutrophils, monocytes and macrophages in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that CLEC4C humanized does not change the overall development, differentiation or distribution of these cell types in the blood. Values are expressed as mean ± SEM.
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Analysis of T cell subpopulation in blood
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Analysis of blood T cell subpopulations by FACS. Blood was isolated from female C57BL/6 and B-hBDCA2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3ε + T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hBDCA2 mice were similar to those in the C57BL/6 mice, demonstrating that BDCA2 humanized does not change the overall development, differentiation or distribution of these T cell subtypes in the blood. Values are expressed as mean ± SEM.
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Summary
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Protein expression analysis:
Human BDCA2 was exclusively detectable in pDCs of B-hBDCA2 mice but not in wild-type mice. Human BDCA2 was not detectable in T cells, B cells and NK cells of B-hBDCA2 mice and wild-type mice.
Leukocytes cell subpopulation analysis：
BDCA2 humanized does not change the overall development, differentiation or distribution of immune cell types in spleen, lymph node and blood.
