Analysis of T cell subpopulation in spleen

Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-Tg(hLILRB1/hLILRB4) mice (n=3, 8-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 TCRβ+ 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 B-Tg(hLILRB1/hLILRB4) mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hLILRB1 and hLILRB4 does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.

Analysis of leukocytes cell subpopulation in thymus

Analysis of thymus T cell subpopulations by FACS. Thymocytes were isolated from female C57BL/6 and B-Tg(hLILRB1/hLILRB4) mice (n=3, 8-week-old). Flow cytometry analysis of the thymocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ 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 B-Tg(hLILRB1/hLILRB4) mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hLILRB1 and hLILRB4 does not change the overall development, differentiation or distribution of these T cell subtypes in thymus. Values are expressed as mean ± SEM.

Analysis of leukocytes cell subpopulation in bone marrow

Analysis of bone marrow T cell subpopulations by FACS. Bone marrow leukocyte were isolated from female C57BL/6 and B-Tg(hLILRB1/hLILRB4) mice (n=3, 8-week-old). Flow cytometry analysis of the bone marrow leukocyte was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ 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 B-Tg(hLILRB1/hLILRB4) mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hLILRB1 and hLILRB4 does not change the overall development, differentiation or distribution of these T cell subtypes in bone marrow. Values are expressed as mean ± SEM.

Analysis of leukocytes cell subpopulation in blood

Analysis of blood leukocyte subpopulations by FACS. Blood leukocytes were isolated from female C57BL/6 and B-Tg(hLILRB1/hLILRB4) mice (n=3, 8-week-old). Flow cytometry analysis of the blood leukocytes 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, dendritic cells, granulocytes, monocytes and macrophages in B-Tg(hLILRB1/hLILRB4) mice were similar to those in the C57BL/6 mice, demonstrating that LILRB1 and LILRB4 humanized does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.

Analysis of T cell subpopulation in blood

Blood routine test in B-Tg(hLILRB1/hLILRB4) mice

Relative LILRB4 mRNA expression in B-Tg(hLILRB1/hLILRB4) mice

Relative LILRB4 mRNA expression. Heart、liver、lung and kidney from female C57BL/6 and B-Tg(hLILRB1/hLILRB4) mice (n=3, 9 week-old) was collected and analyzed for q-PCR. Human LILRB4 mRNA was detectable in heart、liver、lung and kidney from B-Tg(hLILRB1/hLILRB4) mice but not wild-type mice. Values are expressed as mean ± SEM.