Figure 1.

Dectin‐1‐mediated cross‐priming of CD8⁺ T cells in vitro requires Card9 signaling in DCs. (A) BM‐derived DCs (BMDCs) from WT and Card9‐deficient (Card9^−/−) donor mice were stimulated with increasing concentrations of Curdlan or the TLR9 ligand CpG 1826. CD80 expression on BMDCs was analyzed by flow cytometry. An asterisk without brackets indicates comparison to the WT “No stim.” condition. (B) BMDCs from WT, Card9^−/−‐deficient, Dectin‐1‐deficient (Dectin‐1^−/−), and MyD88‐deficient (MyD88^−/−) donor mice were stimulated as described above and were cocultured with magnetically purified, CFSE‐labeled CD8⁺ OT‐I T cells in the presence of OVA protein. IFN‐γ release into the supernatant was analyzed by ELISA. (C–E) BMDCs from WT, Dectin‐1^−/−, or ASC‐deficient (ASC^−/−) mice were stimulated as described above and (C) IL‐1β release was determined by ELISA. (D) After coculture with CD8⁺ OT‐I T cells in the presence of OVA protein, T‐cell proliferation was determined by flow cytometry and (E) IFN‐γ release into the supernatant was analyzed by ELISA. (A, B, D, and E) Data are shown as mean ± SEM of at least triplicate samples and are representative of at least two independent experiments. Data in (C) give mean values ± SEM of n = 4 individual samples (CpG stimulation in WT and Card9^−/−, n = 7) that were pooled from four independent experiments. ^* p < 0.05, ** p < 0.01, *** p < 0.001; one‐way ANOVA with Bonferroni posttests.