Figure 3.

DCs from PPs Instruct Mincle- and Syk-Dependent Th17 Differentiation

(A–C) Naive OT-II T cells were co-cultured for 3 days with dome CD11b⁺ DCs, CD8α⁺ DCs, LysoMacs, or LysoDCs from the indicated genotypes, sorted from PPs (1:1 ratio), and loaded with OVA_323–339 peptide. (A) IL-17 secretion by ELISA. Each dot represents an independent co-culture where myeloid cells were from different mice from two independent experiments. (B and C) Representative FACS plots of IL-17 (B) and IL-22 (C) intracellular staining after OTII re-stimulation.

(D) Il6, Il23a, Tgfb, and Il12b transcripts in PPs of the indicated genotypes by qPCR; they were normalized to Gapdh.

(E and F) Analysis of IL-6 (E) and IL-12p40 (F) intracellular staining in CD11c⁺MHC-II⁺CD19⁻ cells from PPs in the indicated genotypes. Shown on the left are representative histograms. On the right is MFI of staining.

(G) ELISA of IL-6 and IL-23 production by sorted GM-DCs from WT and Mincle-deficient (Clec4e^−/−) mice untreated (medium) or stimulated with gut microbiota (10:1 DC ratio) for 12 h.

Data represent two independent pooled experiments (A) or one representative experiment of at least two performed (B–G). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 (A: one-way ANOVA and Bonferroni post hoc test; D, E, and G: unpaired two-tailed Student’s t test). See also Figure S3.