Analysis of immune cell subpopulations in spleen

Analysis of splenic immune cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-Rag2 KO 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. Compared to cells in wild-type C57BL/6 mice, percent of T cells and B cells were significantly decreased, while percent of all the other cells, including NK cells, dendritic cells, granulocytes, monocytes and macrophages in B-Rag2 KO mice were  significantly increased. The results demonstrate that knockout of Rag2 gene has blocked B and T cell development and differentiation in spleen. Values are expressed as mean ± SEM.

Analysis of T cell subpopulations in spleen

Analysis of immune cell subpopulations in thymus

Analysis of T cell subpopulations in thymus

Analysis of blood immune cell subpopulations by FACS. Blood were isolated from female C57BL/6 and B-Rag2 KO mice (n=3, 6-week-old). Flow cytometry analysis of the blood cells was performed to assess immune cell 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. Compared to cells in wild-type C57BL/6 mice, percent of total T cells, CD4+ T cells, CD8+ T cells and B cells were significantly decreased, while percent of NK cells, monocytes and macrophages in B-Rag2 KO mice were significantly increased. Percent of dendritic cells and granulocytes in B-Rag2 KO mice were similar to those in wild-type C57BL/6 mice. The results demonstrate that knockout of Rag2 gene has blocked B and T cell development and differentiation in blood. Values are expressed as mean ± SEM.

Analysis of T cell subpopulations in blood

Analysis of blood T cell subpopulations by FACS. Blood were isolated from female C57BL/6 and B-Rag2 KO mice (n=3, 6-week-old).  Flow cytometry analysis of the blood cells was performed to assess T cell 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. Percent of CD4+ T cells, CD8+ T cells and Tregs in B-Rag2 KO mice were significantly decreased compared to those in the C57BL/6 mice, demonstrating that knockout of Rag2 gene has blocked T cell development and differentiation in blood. Values are expressed as mean ± SEM.

B-Rag2 KO mice exhibit the significantly reduced lymph node size and weight