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. 2017 Feb 24;46(2):301–314. doi: 10.1016/j.immuni.2017.01.010

Figure 2.

Figure 2

nBreg Cells Regulate Th Cell Polarization Directly or Indirectly via pDCs

(A–C) Neonatal naive CD4+ T cells were activated by anti-CD3 + anti-CD28 and cultured with 10 ng/mL IL-12 (Th1) or without (Th0), alone or in co-culture with HRSV-activated nBreg cells for 6 days. FACS plots (A) and mean frequencies (B) of TNF-α-, IL-2-, IFN-γ-, IL-13-, IL-17-, or IL-22-secreting cells, as determined by intracellular staining for five donors (ANOVA test). Quantification of IFN-γ (C) in the supernatants of the same co-cultures, as determined by ELISA (n = 4 donors; paired t test was used for comparison).

(D–F) Neonatal pDCs were stimulated with HRSV-A either alone or in co-culture with nBreg cells in the presence of neutralizing anti-IL-10 or control antibody (Ctrl) for 48 hr. pDCs were FACS purified again before being used in Th cell differentiation assay. Intracellular IFN-γ, IL-4, IL-17, and IL-22 expression was analyzed by FACS (D and E) and secreted IFN-γ analyzed in the supernatants by ELISA (F).

Results are representatives of three experiments. Results are expressed as the means ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗p < 0.001, and NS for non-significant.