Figure 4.
CD45RBhi-Derived Foxp3+ Cells Are Increased in the Colon after Naive T Cell Transfer into Il23a−/−Rag1−/− Mice
(A) Frequency of Foxp3+ cells among CD4+ T cells from spleen, MLN, and colonic LPL from IL-23-deficient or -sufficient Rag1−/− recipients transferred with CD4+ CD25− CD45RBhi naive T cells. Each point represents an individual mouse; data are pooled from two independent experiments.
(B) Design of the congenic-transfer experiment. A mixture of 99% sorted CD45.1+ CD4+ CD25− CD45RBhi naive T cells and 1% CD45.2+ CD4+ CD25+ CD45RBlow regulatory T cells was injected into Rag1−/− recipients. FACS plots show representative Foxp3 staining of sorted naive (left) and regulatory (right) populations, gated on CD4+ cells.
(C) Approximately 2 months after transfer, cells from spleen, MLN, and colonic LP were stained for Foxp3 and the congenic marker CD45.2. Left: Representative FACS plot showing Foxp3 versus CD45.2 expression in the spleen of transferred Il23a−/−Rag1−/−. The plot is gated on CD3+ CD4+ cells. Right: Percentage of CD45.2+ Foxp3+ cells in the CD3+ CD4+ population in the spleen, MLN, and LP of transferred IL-23-sufficient or -deficient Rag1−/− recipients. Each point represents an individual mouse.
(D) Percentage of CD45.2− Foxp3+ cells in the CD3+ CD4+ population of the mice analyzed in (C). ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ns, not significant.