Figure 3.

Heligmosomoides polygyrus antigens induce stronger Th1 and reduced Th2 responses in the intestinal draining lymph nodes compared to the footpad. (A) 20 µg of H. polygyrus ES antigen (HES) was injected in the footpad or ileal subserosa. IFN-γ⁺ and IL-4⁺ CD4⁺ T cell responses were analyzed 5 days after injection in the popliteal (pLN) or mesenteric (MLN) lymph nodes (n = 5 mice per group, combined data from two independent experiments; mean ± SEM; unpaired t-tests compare LN responses within each experimental group; ***p ≤ 0.001). (B) Ratio of IFN-γ⁺ and IL-4⁺ CD4⁺ T cell responses after HES injection as shown in (A) (mean ± SEM; unpaired t-test compares LN responses; **p ≤ 0.01). (C) From the same experiments, LN cells were restimulated with HES or media for 3 days in vitro and Th1 and Th2 cytokines were measured by ELISA (mean ± SEM; unpaired t-tests compare LN responses within each experimental group; *p ≤ 0.05, ***p ≤ 0.001). (D) Ratio of IFN-γ and IL-4 cytokine levels after HES restimulation as shown in (C) (mean ± SEM; unpaired t-test compares LN responses; **p ≤ 0.01, ***p ≤ 0.001). (E) Worm counts from the upper half of the small intestine (upper SI), lower half of the small intestine (lower SI) and the colon 17 days after infection with 200 L3 H. polygyrus larvae (n = 4 mice, representative of two independent experiments; mean ± SEM; unpaired t-tests compare worm counts to upper SI numbers; ***p ≤ 0.001). (F, G) Mice were infected with 200 L3 H. polygyrus larvae by oral gavage and individual MLNs were collected after 17 days. Frequency and number of IFN-γ, IL-4, and IL-13 producing CD4⁺ T cells in the individual MLNs are shown (n = 5 mice per group, representative of two independent experiments; mean ± SEM; ordinary one-way ANOVA followed by Holm–Sidak’s multiple comparisons test compare LN responses to naïve controls; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001).