Panel A: Flow cytometry plots showing H h-specific C D 4 positive T cells in mesenteric lymph nodes (m L N) and large intestine (L I) of mice treated with I g G 2 a slash 2 b or C D 80 slash C D 86. The axes are labeled F O X P 3-F I T C and R O R gamma t-P E-e F 610. Panel B: Bar graph showing the percentage of R O R gamma t positive Treg cells in m L N of mice treated with I g G 2 a slash 2 b or C D 80 slash C D 86. The y-axis is labeled percent R O R gamma t positive Tregs of H h-specific C D 4 positive T cells. Panel C: Bar graph showing the percentage of R O R gamma t positive Treg cells in L I of mice treated with I g G 2 a slash 2 b or C D 80 slash C D 86. The y-axis is labeled percent R O R gamma t positive Tregs of H h-specific C D 4 positive T cells. Panel D: Flow cytometry plots showing H h-specific R O R gamma t positive Treg cells in m L N and L I of different mouse models. The axes are labeled F O X P 3-F I T C and R O R gamma t-P E-e F 610. Panel E: Bar graph showing the percentage of H h-specific R O R gamma t positive Treg cells in m L N and L I of different mouse models. The y-axis is labeled percent R O R gamma t positive Tregs of H h-specific C D 4 positive T cells. Panel F: Bar graph showing the number of H h-specific R O R gamma t positive Treg cells in m L N and L I of different mouse models. The y-axis is labeled H h-specific R O R gamma t positive Tregs. Panel G: Flow cytometry plots showing endogenous R O R gamma t positive Treg cells in m L N and L I of different mouse models. The axes are labeled F O X P 3-F I T C and R O R gamma t-P E-e F 610. Panel H: Bar graph showing the percentage of endogenous R O R gamma t positive Treg cells in m L N and L I of different mouse models. The y-axis is labeled percent R O R gamma t positive Tregs of C D 4 positive T cells. Panel I: Bar graph showing the number of endogenous R O R t positive Treg cells in m L N and L I of different mouse models. The y-axis is labeled R O R gamma t positive Tregs.
B7 costimulation antagonizes RORγt + Treg cell responses in the gut. (A–C) (A) Flow cytometry plots of Hh-specific CD4+ T cells (mLN, top panel; LI, bottom panel) and frequency of RORγt+ Treg cells in the (B) mLN and (C) LI of C57BL/6J WT mice treated with IgG2a/2b (n = 11) or αCD80/αCD86 (n = 11). RORγt+ Treg cells were gated as CD4+ Foxp3+ RORγt+ T cells, and this strategy was applied consistently in all experiments. (D–F) (D) Flow cytometry plots, (E) frequency, and (F) cell number of Hh-specific RORγt+ Treg cells in the mLN (top panel) and LI (bottom panel) of Cd80−/−Cd86−/−; CD80Tgfl/fl mice (n = 4), Cd80−/−Cd86−/− mice (n = 5), and RORγtCreCd80−/−Cd86−/−; CD80Tgfl/fl mice (n = 5). (G–I) (G) Flow cytometry plots, (H) frequency, and (I) cell number of endogenous RORγt+ Treg cells in the mLN (top panel) and LI (bottom panel) of Cd80−/−Cd86−/−; CD80Tgfl/fl mice (n = 4), Cd80−/−Cd86−/− mice (n = 5), and RORγtCreCd80−/−Cd86−/−; CD80Tgfl/fl mice (n = 5). The data in Fig. 1 are pooled from three independent experiments. The data are shown as means ± SEM; the statistics shown in E, F, H, and I were obtained by ordinary one-way ANOVA with Tukey’s multiple comparisons test. ns, not significant; *P < 0.05; **P < 0.01; ****P < 0.0001.