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. 2014 Dec 4;9(12):e114516. doi: 10.1371/journal.pone.0114516

Figure 8. Regulation of DC-promoted Th cell responses to β-glucan by IFN-γ.

Figure 8

(A) Human monocyte-derived DCs, unprimed (DCs) or primed overnight with IFN-γ (IFN-γ-DCs), were washed extensively and cultured in the presence or absence of β-glucan for 24 h. Culture supernatants (spn) were used to polarize freshly isolated naïve CD45RO-CD4+ T lymphocytes in presence of anti-CD3 and anti-CD28 antibodies. After 4 d, T cells were washed and stimulated with anti-CD3 and PMA for further 18 h. As controls, naïve T cells were also differentiated with optimal combinations of immunoregulatory cytokines into Th17, Th22, and Th1 cells as described in Materials and Methods S1 in File S2. Cytokine secretion was measured by ELISA. Results are mean ± SEM using monocyte-derived DC supernatants from 4 donors, each tested on T cells from 4 different donors. Statistics: Wilcoxon signed-rank test. (B) Human monocyte-derived DCs, unprimed or primed overnight with IFN-γ were stimulated or not for 24 h with β-glucan, IL-1RA (250 µg/ml) and/or IL-1β as indicated. Protein secretion in the culture supernatants was measured by ELISA. Results are mean ± SEM, n = 6 to 9. Statistics: paired t-test when measurements were uncensored or Tobit model analysis in presence of left-censored data.