Figure 7.

An Eomes-dependent genetic program controls the location of CD8⁺ T reg cells. (A–C) The expression of integrin α1 (A), integrin α4 (B), and CXCR5 (C) on pregated CD8⁺ T reg cells are shown. Pooled results from six independent experiments are shown (A–C). (D) Schematic of intravascular labeling of CD8⁺ T cells. (E) Representative FACS plot of pregated CD8⁺ T reg cells (seft) and pool results from three independent experiments (right) are shown. (F) Representative images of WT and Eomes^−/− spleen (n = 3). Dashed circles provide examples of follicles. White arrows indicate CD8⁺ T cells outside follicles. (G) The ratio of CD8⁺ T cell number outside vs. inside follicles. Each number was calculated from an area of 2.39 mm² containing two to seven follicles. Representative (F) or pooled results (G) from two independent experiments are shown with a total of 20 WT and 20 Eomes^−/− follicles analyzed. All mice are 3–6 mo of age. Each symbol represents the results from an individual animal. N.S., not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA with Tukey multi-comparison posttest (E) or Student’s t test (A, B, C, and G). WT, n = 12 (A), 18 (B), 34 (C), or 7 (E); Eomes^−/−, n = 7 (A), 11 (B), 17 (C), or 5 (E); Tgfbr2^−/−, n = 5 (A), 10 (B), or 17 (C); Tgfbr2^−/−Eomes^−/−, n = 6 (A), 8 (B), 18 (C), or 3 (E).