Depletion of either CD8+T cells or the pro-inflammatory cytokine IFN-γ does not affect morbidity and mortality in ΔTreg SSP mice. (A) Representative FACS plots showing CD4+ T cell depletion in the colonic lamina propria (cLP; top) and spleen (bottom) of ΔTreg SSP mice. (B)Salmonella CFUs measured in the spleen of ΔTreg SSP and CD4-depleted ΔTreg SSP at 28 dpi (n = 8–15). (C) Schematic representation of the experimental design. F1-Foxp3DTR mice (n = 10) were infected with Salmonella Typhimurium SL1344 (108 CFUs; STm), treated with streptomycin to induce SSP at 14 dpi, and injected (i.p.) with DT and anti-CD8β/isotype antibodies to deplete CD8+ T cells, as indicated. (D) Survival curve (left) and percentage of body weight variation (right) of ΔTreg SSP mice treated with anti-CD8β (turquoise circle) or isotype antibodies (orange circle) starting at day 21 after infection (n = 10). (E) FITC-dextran (ng/ml) measured in the serum of ΔTreg SSP and CD8-depleted ΔTreg SSP at 28 dpi (n = 6–10). (F)Salmonella CFUs measured in the fecal content (left) and spleen (right) of ΔTreg SSP and CD8-depleted ΔTreg SSP at 28 dpi (n = 10). (G) Schematic representation of the experimental design. F1-Foxp3DTR mice (n = 10) were infected with Salmonella Typhimurium SL1344 (108 CFUs; STm), treated with streptomycin to induce SSP at 14 dpi, and injected (i.p.) with DT and anti-CD4 and CD8β or isotype antibodies to deplete CD4+ and CD8+ T cells, as indicated. (H) Survival curve (left) and percentage of body weight variation (right) of ΔTreg SSP mice treated with anti-CD4 and CD8β (magenta circle) or isotype antibodies (orange circle) starting at day 21 after infection (n= 10–15). A log-rank (Mantel–Cox) test was used to compare survival probability between the two groups. (I) FITC-dextran (ng/ml) measured in the serum of ΔTreg SSP and CD4-CD8-depleted ΔTreg SSP at 28 dpi (n = 6–10). (J)STm CFUs measured in the fecal content (left) and spleen (right) of ΔTreg SSP (orange) and CD4- and CD8-depleted ΔTreg SSP (magenta) at 28 dpi (n = 10). (K) Survival curve (top) and percentage of body weight variation (bottom) of ΔTreg SSP mice treated with anti-IFN-γ (lilac circle) or isotype antibodies (orange circle) starting at day 21 after infection (n = 10). (L)STm CFUs measured in the fecal content (top) and spleen (bottom) of ΔTreg SSP mice treated with anti-IFN-γ (lilac) or isotype antibodies (orange) at 28 dpi (n = 10). Results are representative of at least two independent experiments and presented as means ± SEM. Normality was assessed by the D’Agostino–Pearson test. The Mann–Whitney U (B, E, F, I, J, and L) test was used to compare two groups (****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05, ns = not significant). Two-way ANOVA (D, H, and K) followed by post hoc Sidak’s test was performed for multiple groups comparisons. A log-rank (Mantel–Cox) (H) test was used to compare survival probability between the two groups.
Figure S5.

Depletion of either CD8 + T cells or the pro-inflammatory cytokine IFN-γ does not affect morbidity and mortality in ΔTreg SSP mice. (A) Representative FACS plots showing CD4+ T cell depletion in the colonic lamina propria (cLP; top) and spleen (bottom) of ΔTreg SSP mice. (B)Salmonella CFUs measured in the spleen of ΔTreg SSP and CD4-depleted ΔTreg SSP at 28 dpi (n = 8–15). (C) Schematic representation of the experimental design. F1-Foxp3DTR mice (n = 10) were infected with Salmonella Typhimurium SL1344 (108 CFUs; STm), treated with streptomycin to induce SSP at 14 dpi, and injected (i.p.) with DT and anti-CD8β/isotype antibodies to deplete CD8+ T cells, as indicated. (D) Survival curve (left) and percentage of body weight variation (right) of ΔTreg SSP mice treated with anti-CD8β (turquoise circle) or isotype antibodies (orange circle) starting at day 21 after infection (n = 10). (E) FITC-dextran (ng/ml) measured in the serum of ΔTreg SSP and CD8-depleted ΔTreg SSP at 28 dpi (n = 6–10). (F)Salmonella CFUs measured in the fecal content (left) and spleen (right) of ΔTreg SSP and CD8-depleted ΔTreg SSP at 28 dpi (n = 10). (G) Schematic representation of the experimental design. F1-Foxp3DTR mice (n = 10) were infected with Salmonella Typhimurium SL1344 (108 CFUs; STm), treated with streptomycin to induce SSP at 14 dpi, and injected (i.p.) with DT and anti-CD4 and CD8β or isotype antibodies to deplete CD4+ and CD8+ T cells, as indicated. (H) Survival curve (left) and percentage of body weight variation (right) of ΔTreg SSP mice treated with anti-CD4 and CD8β (magenta circle) or isotype antibodies (orange circle) starting at day 21 after infection (n= 10–15). A log-rank (Mantel–Cox) test was used to compare survival probability between the two groups. (I) FITC-dextran (ng/ml) measured in the serum of ΔTreg SSP and CD4-CD8-depleted ΔTreg SSP at 28 dpi (n = 6–10). (J)STm CFUs measured in the fecal content (left) and spleen (right) of ΔTreg SSP (orange) and CD4- and CD8-depleted ΔTreg SSP (magenta) at 28 dpi (n = 10). (K) Survival curve (top) and percentage of body weight variation (bottom) of ΔTreg SSP mice treated with anti-IFN-γ (lilac circle) or isotype antibodies (orange circle) starting at day 21 after infection (n = 10). (L)STm CFUs measured in the fecal content (top) and spleen (bottom) of ΔTreg SSP mice treated with anti-IFN-γ (lilac) or isotype antibodies (orange) at 28 dpi (n = 10). Results are representative of at least two independent experiments and presented as means ± SEM. Normality was assessed by the D’Agostino–Pearson test. The Mann–Whitney U (B, E, F, I, J, and L) test was used to compare two groups (****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05, ns = not significant). Two-way ANOVA (D, H, and K) followed by post hoc Sidak’s test was performed for multiple groups comparisons. A log-rank (Mantel–Cox) (H) test was used to compare survival probability between the two groups.

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