Panel A: Flow cytometry plots showing the distribution of H h-specific C D 4 positive T cells in the mesenteric lymph nodes (m L N) and large intestine (L I) of H 2-A b 1 f l slash f l mice and R O R gamma t C r e H 2-A b 1 f l slash f l mice. The x-axis represents R O R gamma t-P E-e F 610 and the y-axis represents F O X P 3-F I T C. Panel B: Bar graphs showing the frequency of H h-specific R O R gamma t positive Treg cells in m L N and L I. The y-axis represents the percentage of R O R gamma t positive Treg cells. Panel C: Bar graphs showing the frequency of H h-specific T H 17 cells in m L N and L I. The y-axis represents the percentage of T H 17 cells. Panel D: Flow cytometry plots showing the distribution of H h-specific C D 4 positive T cells in m L N and L I of H 2-A b 1 f l slash f l mice and R O R gamma t C r e H 2-A b 1 fl slash fl mice treated with anti-I L-10 R slash C T L A 4-I g. The x-axis represents R O R gamma t-P E-e F 610 and the y-axis represents F O X P 3-F I T C. Panel E: Bar graphs showing the frequency of H h-specific R O R gamma t positive Treg cells in m L N and L I. The y-axis represents the percentage of R O R gamma t positive Treg cells. Panel F: Bar graphs showing the cell number of H h-specific R O R gamma t positive Treg cells in m L N and L I. The y-axis represents the cell number. Panel G: Bar graph showing the frequency of H h-specific T H 17 cells in L I. The y-axis represents the percentage of T H 17 cells. Panel H: Flow cytometry plots showing the distribution of T N F positive H h-specific T H 17 cells in L I. The x-axis represents C D 4-B V 650 and the y-axis represents T N F-A F 480. Panel I: Bar graph showing the frequency of T N F positive H h-specific T H 17 cells in L I. The y-axis represents the percentage of T N F positive T H 17 cells. Panel J: Images of the large intestine (L I) showing typhlocolitis in H 2-A b 1 f l slash f l mice and R O R gamma t C r e H 2-A b 1 fl slash fl mice. Panel K: Images of the enlarged mesenteric lymph nodes (m L N) in H 2-A b 1 f l slash f l mice and R O R gamma t C r e H 2-A b 1 f l slash f l mice. Panel L: Bar graph showing the cell number of neutrophil infiltration in the L I. The y-axis represents the cell number.
RORγt + APCs are required for CTLA4-Ig–driven immune tolerance. (A–C) (A) Flow cytometry plots of the Hh-specific CD4+ T cells (mLN, top panel; LI, bottom panel) and frequencies of (B) Hh-specific RORγt+ Treg cells and (C) Hh-specific TH17 cells in the mLN (top panel) and LI (bottom panel) of H2-Ab1fl/fl mice (n = 6) and RORγtCreH2-Ab1fl/fl mice (n = 6). (D–F) (D) Flow cytometry plots of the Hh-specific CD4+ T cells (mLN, top panel; LI, bottom panel), (E) frequency, and (F) cell number of Hh-specific RORγt+ Treg cells in the mLN (top panel) and LI (bottom panel) of H2-Ab1fl/fl mice (n = 4) and RORγtCreH2-Ab1fl/fl mice (n = 5). (G) Frequency of Hh-specific TH17 cells in LI of the mice in D. (H and I) (H) Flow cytometry plots and (I) frequency of TNF+ Hh-specific TH17 cells in LI of the mice in D. (J–L) Images of (J) LI showing typhlocolitis, (K) enlarged mLNs, and (L) cell number of neutrophil infiltration in the LI of the mice in D. The data in A–C are pooled from two independent experiments. The data in D–L are representative of two independent experiments. The data are shown as means ± SEM; the statistics shown in B, C, E, F, G, I, and L were obtained by unpaired Student’s t test (two-tailed). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.