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.

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.

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.

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.

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.

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.

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.