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. 2017 Jul 28;13(7):e1006539. doi: 10.1371/journal.ppat.1006539

Fig 1. Schistosome eggs and SEA (soluble egg antigen) drive development of Breg cells in vivo.

Fig 1

C57BL/6 mice were i.p. injected with two doses of 5000 S. mansoni eggs or 100 μg SEA in PBS, or PBS as control. At day 14, CD19+ MACS-isolated splenic B cells were restimulated with SEA (20 μg/ml) for 2 days. (A) Cytokine concentration in culture supernatants as determined by ELISA. (B, C) Representative FACS plots (B) and summary (C) for intracellular IL-10 expression of B cells after addition of Brefeldin A to the last 4 hours of the culture. (D) Mean fluorescence intensity of CD40 and CD86 expression. (E-G) SEA-restimulated B cells were co-cultured for 4 days with CD25-depleted CD4 T cells. Frequency of CD25+Foxp3+ Treg cells after co-culture in representative FACS plots (E) and summary (F) is shown. (G) IL-10 concentration in culture supernatants after co-culture. Summary of 4 experiments with N = 12–16 (A-D) or 2 experiments with N = 8–9 (E-G). Significant differences by Mann-Whitney test are indicated with * p < 0.05, ** p < 0.01, *** p < 0.001.