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. 2016 May 13;5(7):e1180485. doi: 10.1080/2162402X.2016.1180485

Figure 4.

Figure 4.

TRAPs induce IL-10-producing B cell via TLR2-MyD88-NF-κB dependent signaling pathway. (A) B cells purified from WT, TLR2-, TLR4-, or MyD88 deficient mice were stimulated with TRAPs (3 μg/mL) for 72 h. Frequency of IL-10-producing B cells was determined by flow cytometry. (B and C) B cells purified from WT mice were pre-treated with NF-κB inhibitor Bay11–7082 at different concentrations for 60 min, and then stimulated with TRAPs (3 μg/mL) for 72 h. Frequency of IL-10+ B cells was assessed by flow cytometry (B). IL-10 levels in supernatants were measured by ELISA (C). (D) B cells purified from WT, TLR2-, TLR4-, or MyD88 deficient mice were stimulated with TRAPs (3 μg/mL) for 1 h, and the expression of pNF-κB p65 was determined by flow cytometry. (E) CFSE-labeled CD4+ T or CD8+ T cells purified from WT mice were stimulated with plate-bound anti-CD3/anti-CD28 mAb and were either cultured alone (T cells only) or were co-cultured with TRAP-induced B cells (3 μg/mL) from WT, TLR2-, TLR4-, and MyD88 deficient mice at ratio of 1:1 for 4 d. T cell proliferation was evaluated by flow cytometry. Data (mean ± s.e.m) are representative of three independent experiments. ** p < 0.01, *** p < 0.001 by unpaired t-test.