Cell-Intrinsic Role for NF-kappa B-Inducing Kinase in Peripheral Maintenance but Not Thymic Development of Foxp3+ Regulatory T Cells in Mice
Figure 4
Decreased peripheral Foxp3+ Tregs in NIK KO BM chimeric mice is cell-intrinsic.
Single cell suspensions of nucleated splenocytes (A-F) or thymocytes (G) from NIK KO (CD45.2) + WT (CD45.1xCD45.2) F1 or WT littermate (CD45.2) + WT (CD45.1xCD45.2) F1 mixed BM chimeric mice were stained with the indicated fluorescently labeled antibodies. A, Gating scheme for identifying cells in mixed BM chimeras. CD45.1 + CD45.2- cells (ungated) are radioresistant host cells. Numbers indicate percent of gated cells. B and C, Quantitation of percent and absolute number of donor CD45.2 + CD45.1-Foxp3+ cells. D and E, Quantitation of percent and absolute number of donor CD45.2 + CD45.1-CD4+ cells. These data compare NIK KO cells with WT littermate control cells. Because WT (CD45.1xCD45.2) F1 cells appear to have a slight, but consistent advantage over CD45.2 + CD45.1- WT littermates in terms of both overall proportion and proportion of Foxp3+ cells, comparing NIK KO T cells to WT littermate T cells is the most conservative and appropriate comparison. When we compare NIK KO T cells to WT (CD45.1xCD45.2) F1 T cells, the differences are even greater. F, CD25 expression on NIK KO (bold line) versus WT (dashed line) Foxp3+ splenocytes. G, Quantitation of percent of NIK KO and WT SP4 and Foxp3+ thymocytes in mixed BM chimeras. Data in bar graphs represent mean ± SEM of n=3-4 mice per group in one representative experiment of two.