T. muris infection chronically increases cecal IFN-γ⁺ T cells. (A–D) C57BL/6 breeders were orally gavaged with 10⁶ isolated protozoans. (A and B) Ceca (A) and colons (B) of infected pups and control pups from noninfected breeders were harvested. Cells were isolated and restimulated for 4 h with PMA and ionomycin, in the presence of protein transport inhibitor cocktail, before flow cytometry analysis. Data from two experiments were pooled with three to six mice per group. (C) Proximal colon explants were cultured for 24 h, and supernatants were analyzed by ELISA for IL-12/IL-23p40 and IL-18 protein. Data from two experiments were pooled with six to seven samples per group. (D) Small intestines were fixed, stained for tufts cells (DCLK1⁺DAPI⁺), and quantified. Data from two experiments were pooled with six mice per group. (A–D) Mean ± SEM is shown with *, P ≤ 0.05 using an unpaired Student’s t test.