FIG. 6.

Cytokine subunit mRNA accumulation and T-cell differentiating capacity of cDC subsets are altered during the course of infection. (A) Splenic CD11c^hi DC subsets were isolated from naïve and LD-infected mice (2 × 10⁷) at different times (5 h, 24 h, day 14 [d14], and day 28 [d28]). Ex vivo analysis of IL-12/23p40, IL-23p19, and IL-27p28 mRNA accumulation in CD4⁺ (white bars), CD8α⁺ (gray bars), and DN (black bars) DC were examined by real-time RT-PCR. Target genes were normalized against HPRT. (B) Data from panel A are summarized as changes in the accumulation of cytokine mRNA from infected DC subsets compared to their naïve counterpart. (C) Sorted DC subsets from naïve and infected mice (5 h, day 14, and day 28) were cultured with purified naïve DO11.10 T cells in the presence of 0.1 μg/ml OVA peptide. Expanded cells were subsequently restimulated with whole spleen cells and 2 μg/ml OVA peptide and stained for IFN-γ and IL-4. Plots were gated on CD4⁺/KJ1-26⁺. The numbers indicate the percentage of events within the two quadrants. Data are representative of three independent experiments.