FIG 5.

Trophic forms reduce dendritic cell-dependent proliferation of CD4⁺ T cells. Treatment with trophic forms reduces CD4⁺ T cell proliferation in response to coculture with allogeneic BMDCs in a mixed-lymphocyte reaction. (A) BMDCs generated from BALB/cJ mice were pretreated with trophic forms or mixed P. murina organisms for 1 h. CFSE-stained splenocytes from uninfected C57BL/6 mice were then added to the wells and cocultured for 6 days. Flow cytometry was used to identify the CD4⁺ T cells and evaluate their proliferation. (B to E) BMDCs stimulated with trophic forms induce less proliferation and cytokine protein expression in CD4⁺ T cells than cells stimulated with mixed P. murina organisms. BMDCs and CFSE-stained CD4⁺ T cells from adult BALB/cJ mice were incubated with trophic forms or mixed P. murina organisms for 6 days. Flow cytometry was used to evaluate proliferation of CD4⁺ T cells (B). ELISA was used to evaluate the concentrations of the cytokines IFN-γ (A and C), IL-13 (D), and IL-17A (E) in the supernatant. Data represent the means ± standard deviations of three biological replicates per group and are representative of at least two separate experiments. One-way ANOVA with Student-Newman-Keuls post hoc test was used to compare the percentage of proliferating CD4⁺ T cells or supernatant cytokine concentration among the groups: *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; n.s., not statistically significant.