FIG 3.

S. mansoni-infected Cd14^−/− mice have enhanced Th2 responses. (A) Splenocytes were prepared from naive and S. mansoni-infected (12 weeks) (Sm) wild-type C57BL/6 (n = 5) and Cd14^−/− mice (n = 5). Cells were then surface stained with antibodies to CD4 and intracellular cytokines IL-4, IL-13, IL-5, IL-10, IL-17, and IFN-γ and analyzed by flow cytometry. The percentages of CD4⁺ T-cell-specific cytokines (among total CD4⁺ T cells) are shown. (B) Splenocytes were stimulated with SEA (25 μg/ml) at 37°C for 72 h, and then supernatants were collected. Cytokine production was assayed by ELISA. Med, medium. (C, D) qRT-PCR was performed specific to cytokines (genes Il-4 and Il-13) and chemokines (genes Ccl17 and Ccl22), as indicated, using total RNA extracted from livers of naive or schistosome-infected mice. Gene expression was analyzed as fold changes over naive controls after normalization with GAPDH. (E) Production of SEA-specific IgG1 and total IgE in the sera of naive or infected mice determined by ELISA. Statistical significance was calculated using one-way ANOVA Tukey's multiple-comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. Results (means ± SEM) represent 3 or more independent experiments.