Panel A: Flow cytometry plots showing the frequency of c-M a f positive cells of H h-specific C D 4 positive T cells. The x-axis represents C D 4-B V 650, and the y-axis represents c-M a f-P E. The bar graph shows a significant increase in c-M a f positive cells with C T L A 4-I g treatment. Panel B: Flow cytometry plots showing the frequency of c-M a f positive cells of H h-specific C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The x-axis represents C D 4-B V 650, and the y-axis represents c-M a f-P E. The bar graph shows a significant increase in c-M a f positive cells with C T L A 4-I g treatment. Panel C: Flow cytometry plots showing the frequency of R O R gamma t positive Treg cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The x-axis represents R O R gamma t-P E-E 610, and the y-axis represents F O X P 3-R I T C. The bar graph shows a significant increase in R O R gamma t positive Treg cells with C T L A 4-I g treatment. Panel D: Bar graph showing the frequency of R O R gamma t positive Treg cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of R O R gamma t positive Treg cells. Panel E: Bar graph showing the frequency of T H 17 cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of T H 17 cells. Panel F: Flow cytometry plots showing the frequency of c-M a f positive cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The x-axis represents C D 4-B V 650, and the y-axis represents c-M a f-P E. The bar graph shows a significant increase in c-M a f positive cells with C T L A 4-I g treatment. Panel G: Bar graph showing the frequency of c-M a f positive cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of c-M a f positive cells. Panel H: Flow cytometry plots showing the frequency of T-bet positive T H 17 cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The x-axis represents C D 4-B V 650, and the y-axis represents T-bet-e 660. The bar graph shows a decrease in T-bet positive T H 17 cells with C T L A 4-I g treatment. Panel I: Bar graph showing the frequency of T-bet positive T H 17 cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of T-bet positive T H 17 cells. Panel J: Bar graph showing the frequency of I F N positive C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of I F N positive C D 4 positive T cells. Panel K: Bar graph showing the frequency of I F N positive I L-17 A positive C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of I F N positive I L-17 A positive C D 4 positive T cells. Panel L: Flow cytometry plots showing the frequency of T N F positive T H 17 cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The x-axis represents C D 4-B V 650, and the y-axis represents T N F-e 450. The bar graph shows a decrease in T N F positive T H 17 cells with C T L A 4-I g treatment. Panel M: Bar graph showing the frequency of T N F positive T H 17 cells of C D 4 positive T cells in the L I of mice treated with I g G 1 F c and C T L A 4-I g. The y-axis represents the percentage of T N F positive T H 17 cells. Panel N: Experimental timeline showing the treatment schedule for 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, including H h colonization, C D 4 positive T cell transfer, anti-I L-10 R injections, and C T L A 4-I g injections.
CTLA4-Ig drives immune regulation in a mouse model of gut inflammation. (A and B) Flow cytometry plots and frequency of c-Maf+ Hh-specific CD4+ T cells in the (A) mLN and (B) LI of C57BL/6J WT mice treated with IgG1 Fc (n = 5) or CTLA4-Ig (n = 4) as shown in Fig. 3. (C–G) (C) Flow cytometry plots and frequencies of endogenous CD4+ T cells in the LI of the mice in Fig. 3, showing (D) RORγt+ Treg cells, (E) TH17 cells, and (F and G) c-Maf+ cells. (H–M) Flow cytometry plots and frequencies of inflammatory endogenous CD4+ T cell subsets in the LI of the mice as shown in Fig. 3, showing (H and I) T-bet+ TH17 cells, (J) IFNγ+ CD4+ T cells, (K) IFNγ+IL-17A+ CD4+ T cells, and (L, M) TNF+ TH17 cells. (N)H2-Ab1fl/fl mice and RORγtCreH2-Ab1fl/fl mice were colonized with Hh on days 0, 2, and 4, followed by Hh-specific naïve CD4+ T cell transfer retro-orbitally on day 7; mice were i.p. injected with anti-IL-10R on days 0, 6, 12, and 18, and were i.p. injected with CTLA4-Ig on days 0, 3, 6, 10, 13, 16, 19, and 22. Mice were euthanized for analysis on day 24. The data in Fig. S3 are representative of two independent experiments. The data are shown as means ± SEM; the statistics shown in A, B, D, E, G, I, J, K, and M were obtained by unpaired Student’s t test (two-tailed). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.