Fig. 5: pDC heterogeneity correlates with distinct cytokine-producing capacity.
From: Tonic type I interferon signaling optimizes the antiviral function of plasmacytoid dendritic cells
a–e, Phenotypic pDC subsets were sorted from the splenocytes pooled from six naive WT mice and cultured (2 × 104 per well) in the presence of TLR agonists, and concentrations of indicated proteins in the supernatants were measured 12–18 h later. For each multiplex panel, all analytes were measured at the same time point. In all panels, bars represent means ± s.d., and symbols represent values from independently sorted biological replicates (single cultures or averages of technical replicates). Note that the rare pDC-A and pDC-C populations were assayed in fewer experiments than for pDC-B. a, Cells were stimulated with CpG-A and IFNα was measured by ELISA. b, Cells were stimulated with CpG-A and indicated analytes were measured by a multiplex bead array. c, Cells were stimulated with ssRNA40 and IFNα was measured by ELISA. d, Cells were stimulated with ssRNA40 and indicated analytes were measured by a multiplex bead array. e, Cells were stimulated with CpG-B and indicated analytes were measured by a multiplex bead array. f, T cell priming by pDC subsets. pDC subsets or CD11chiCD11b+ cDC2 were sorted from pooled spleens of naive B6 WT mice, plated at 3 × 104 per well and co-cultured with 3 × 104 magnetically enriched T cells from OT-II mice in the presence of OVA323–339 peptide. The numbers of OVA-specific (TCR-Vβ-5.1/2+) CD4+ T cells in each well were determined by flow cytometry 64 h later. Control wells were seeded with T cells without DCs with or without OVA. Bars represent means ± s.d.; symbols represent cultures of separately sorted cells from one experiment, representative of three experiments. Statistical significance was analyzed using one-way ANOVA followed by Tukey’s test; *P < 0.05; **P < 0.005; ***P < 0.0005; ****P < 0.00005. Statistics for comparisons with controls not shown: black bars (b–e), gray bars (f). Comparisons with control omitted from statistical analysis: black bar (b). m, mouse; Tc, T cell; Unstim., unstimulated.
