Tα1-induced DCs prime for antifungal Th1/Treg responses in vivo. Patterns of inflammatory/Th/Treg responses 3 days after the infection in mice treated as described in Figure 2 legend. (A) TNF-α/IL-10 levels were assessed by specific ELISA in lung homogenates, and IFN-γ/IL-4 production was assessed in TLN CD4⁺ T cells cocultured with Aspergillus-pulsed DCs. Bars indicate standard errors. TLN CD4⁺CD25⁺ T cells producing IL-10 or TGF-β were numbered by ELISPOT assay. Results are expressed as the mean number of cytokine-producing cells (±SE) per 2 × 10⁵ cells. *P < .05, DC-treated versus untreated mice. **P < .05, Tα1-treated DCs versus untreated DCs. (B) Total RNA was extracted from freshly purified CD4⁺ T cells from TLNs of treated or untreated (None) mice. The expressions of the different mRNAs in each cell population were determined by RT-PCR. The expression of a housekeeping gene, Gapdh mRNA, was used as an internal control. The data shown are representative results of 3 experiments. (C) Phenotypic analysis of cells isolated from lung or TLNs of mice infused or not (None) with different types of DCs, (–) indicating uninfected, untreated mice. CD4⁺ T cells were sequentially reacted with PE-conjugated anti-CD25 (PC61) and FITC-conjugated anti-CD69 (clone HI.2F3) mAbs. Numbers represent the percentage of positive cells over total cells analyzed. Control staining of cells with irrelevant Ab was used to obtain background fluorescence values. Histograms are representative of 1 of 4 independent experiments.