Fig 3. Depletion of Foxp3+ Tregs during T. spiralis infection results in extreme morbidity and delayed helminth expulsion, but the immune kinetics and delayed expulsion seen in mice lacking the TGFβ-activating integrin αvβ8 on DCs are independent of Tregs.

DEREG mice were treated every 2 days with 200 ng diphtheria toxin or PBS (Control) 2 days prior to infection with 300 T. spiralis larvae and examined at the indicated time-points post-infection. (A) Worm burdens from control and DEREG mice at days 7 and 15 following infection. (B) Percentage change in basal start weight in control and DEREG mice during time course of infection, dashed line indicates point of morbidity sacrifice threshold. (C) IL-4, IL-13, IFNγ, IL-6 and IL-17 cytokine levels from T. spiralis antigen-stimulated mLN cells from control and DEREG mice at different time-points post-infection, determined via CBA. Data (n = 4–11 mice per group) are from two independent experiments performed. Wild-type, Itgb8 (CD11c-cre) and Itgb8 (CD11c-cre) mice adoptively transferred with 1x10⁶ Tregs were infected with 300 T. spiralis larvae 2 days following cell transfer and examined at the indicated time-points post-infection. (D) Percentage change in basal start weight in wild-type, Itgb8 (CD11c-cre) and Itgb8 (CD11c-cre) mice adoptively transferred with Tregs during time course of infection. (E) Representative flow cytometry plots for intracellular IL-17 and IL-13 expression in mLN CD4+ T-cells isolated from wild-type, Itgb8 (CD11c-cre) and Itgb8 (CD11c-cre) mice adoptively transferred with Tregs, at day 13 post-infection. (F) Number of IL-17+ CD4 T-cells in the mLN of wild-type, Itgb8 (CD11c-cre) and Itgb8 (CD11c-cre) mice adoptively transferred with Tregs, at day 13 post-infection, assessed via flow cytometry. (G) Worm burdens from wild-type, Itgb8 (CD11c-cre) and Itgb8 (CD11c-cre) mice adoptively transferred with Tregs, at day 13 following infection. Data (n = 4 mice per group) are from two independent experiments performed.*, P<0.05; **, P<0.01; ***, P<0.005; N.S., not significant via Bonferonni’s multiple comparison following ANOVA (C), (F) and (G), and student’s t-test (A), (B) and (D) for indicated comparisons between groups.