Figure 2.

Activation of mDCs via Dectin-1 Inhibits TSLP-Induced Inflammatory Th2 Response of CRTH2⁺ Memory CD4⁺ T Cells. (A) Intracellular staining of cytokines expressed by sorted CRTH2⁺ memory CD4⁺ T cells upon ex vivo stimulation with PMA/ionomycin. The FACS data shown is representative of three independent experiments. (B, C) Sorted mDCs were treated with indicated reagents for 24h before coculture with autologous CRTH2⁺ memory CD4⁺ T cells for 7d followed by intracellular staining of cytokines expressed by T cells. Live singlets were gated for analysis. Representative FACS data (B) and summarized data (C) from different donors (n = 3) are shown. (D) After 5d coculture of mDCs and autologous CRTH2⁺ memory CD4⁺ T cells as in (B), T cells were stimulated by anti-CD3/28 beads for 48h before measuring cytokine production in culture supernatants by multiplex assay. Summarized data from different donors (n = 3) are shown. (E) After 4d or 7d of coculture of mDCs and autologous CRTH2⁺ memory CD4⁺ T cells as in (B), surface CRTH2 expression on CD4⁺ T cells was measured by FACS staining. Summarized data from different donors (n = 3) are shown. Data in (C–E) are represented as mean ± SD. Significance was determined using one-way ANOVA with Tukey multiple comparisons test for (C–E). *p < 0.05, **p < 0.01 for the comparison between groups. ns, not significant.