Role of Protein Kinase C and Nox2-Derived Reactive Oxygen Species Formation in the Activation and Maturation of Dendritic Cells by Phorbol Ester and Lipopolysaccharide

Figure 3

Analysis of intracellular ROS formation upon activation of BM-DCs, of maturation at 24 h after stimulation, and of their T cell stimulatory capacity. (a) 1 × 10⁶ BM-DCs were stimulated with LPS (1.5 μg/ml) or PDBu (10 μM). ROS formation (a) in and (b) expressing CD11c⁺ cells was analyzed by flow cytometry after 15 or 120 min. (c) Maturation was characterized by high MHCII and CD86 expression after 24 h using flow cytometry. (d) T cell stimulatory capacity was compared in a proliferation assay with a constant number of T cells (3 × 10⁵) cocultured with decreasing numbers of differentially stimulated BM-DCs, and on day 4 proliferation of T cells was measured by incorporation of [³H] thymidine. Data are mean ± SEM of (a) 4-5, (b) 4-5, (c) 5, and (d) 3 experiments. ; versus PDBu (a, b); versus unstimulated; ; versus PDBu (c, d).

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