Figure 5.

PKCδ- and Nox2-mediated antigen macropinocytosis regulates DC secretion of T-cell regulatory cytokines. (A–C) Wild-type (WT) (C57BL/6J), PKCδ^−/− and Nox2^y/− BMiDCs were treated with vehicle or PMA in the presence of unlabeled OVA (50 µg/mL) or PMA alone for 24 h. Cell culture supernatants were collected and levels of secreted cytokines determined using LEGENDplex™ bead-based immunoassay assay. Bar graphs show fold changes in cytokine secretion compared to “vehicle + OVA” treatment. Data are representative of three independent experiments and values are expressed as mean ± SD. (D) BMiDCs were treated as described above. The mRNA levels of IL-6 were determined by qRT-PCR. Bar graphs show fold change in the expression of IL-6 compared to “vehicle + OVA” treatment (n = 3) and data represent the mean ± SD. (E–G) WT, PKCδ^−/−, and Nox2^y/− CD11c⁺ splenic iDCs were treated with vehicle + OVA, PMA + OVA, or PMA alone for 24 h. The mRNA levels of cytokines were determined by qRT-PCR. Data are representative of three independent experiments performed in triplicate and represent the mean ± SEM. Bar graphs show fold change in the expression compared to “vehicle + OVA” treatment. *p < 0.05 vs. vehicle + OVA, and ^@p < 0.05 vs. WT PMA + ova.