FIG 2.

DCs from anti-TNF-treated H. capsulatum-infected mice amplify Tregs. (A to C) Mice were infected with 2 × 10⁶ H. capsulatum yeasts i.n. and treated with anti-TNF or isotype control antibody. Lungs were harvested at day 3 p.i., and DCs (CD11c⁺ CD11b⁺ MHC-II⁺) (A and B) and alveolar macrophages (CD11c⁺ CD11b⁻ F4/80⁺) (C) were assessed by FACS. The cells were cocultured with splenic CD4⁺ CD25⁻ Foxp3⁻ T cells from uninfected Foxp3-GFP mice in the presence of 1 μg/ml anti-CD3, 5 μg/ml anti-CD28, 3 ng/ml TGF-β, and 5 ng/ml IL-2. (D) Mice were treated with anti-TNF or isotype control antibody. Lungs were harvested 3 days after treatment, and DCs (CD11c⁺ CD11b⁺ MHC-II⁺) were assessed by FACS. The cells were cocultured as described above. The data represent means and SEM of the results of at least 3 experiments. Lungs of 4 to 8 mice were pooled for each experiment. *, P < 0.05.