Figure 6.

Batf3-dependent migratory CD103⁺ DCs are major constitutive producers of IL-12 both at steady state and during helminth infections. (A) mLN cells from naive yet40 mice were cultured for 6 h in the presence or absence of Brefeldin A. YFP signal and IL-12p40 protein expression by CD103⁺ DCs was determined by flow cytometry. (B) hLN and mLN cells from naive C57BL/6 or yet40 IL-12 reporter C57BL/6 mice were analyzed for YFP expression. (C and E) yet40 IL-12 reporter mice were infected with S. mansoni for 6 wk (C) or 7 d (E) with H. polygyrus and migratory (mDCs; MHCII^hiCD11c^int) and resident (rDCs; MHCII^intCD11c^hi) DC subsets in hLNs (C) and mLNs (E) were analyzed for YFP reporter expression. The gating strategy for mDC and rDC subsets is shown in the three left panels. The far left panel shows hLN (C) or mLN (E) cells from a naive WT mouse and from which CD19⁺ B cells, which can also be CD11c⁺MHCII⁺, have been gated out. Data are concatenated plots from three to four mice per group. (D and F) DC subsets as described in C and E were analyzed for MHC-II expression. (G and H) Bar graphs represent frequency of IL-12p40-producing DC subsets of total cells based on YFP expression within hLNs from naive or S. mansoni-infected mice (G) or mLNs from naive or H. polygyrus-infected mice (H). Pie charts depict relative contribution of each DC subset to YFP expression within (G) hLNs from S. mansoni-infected mice or (H) mLNs from H. polygyrus-infected mice. (D and F–H) Data shown as bar graphs represent mean ± SEM from three to four mice per group. One of two (A and B) or three (C–H) experiments is shown. *, P < 0.05.