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. 2019 Apr 24;10:852. doi: 10.3389/fimmu.2019.00852

Figure 1.

Figure 1

Tonsil B cells express T-bet upon BCR, TLR9, and IFN-γ stimulation in vitro. Tonsil B cells were cultured in vitro for 40 h with combination of antigen, either soluble or presented on a planar lipid bilayer (PLB) or plasma membrane sheet (PMS), CpG, IFN-γ, or IL-12 + IL-18. T-bet expression in naïve (IgD+CD10−), memory (IgD−CD10−), and GC (IgD−CD10+) B cells was analyzed by flow cytometry. (A) Heat map indicating T-bet expression (gMFI) by naïve, memory, and GC B cells of the three individuals (columns) from three representative experiments. The stimulation conditions are as indicated to the right of heat map and are grouped into single, double, triple, and quadruple stimuli. The columns are subgrouped according to the mode of antigen presentation either attached to PLB or PMS or soluble. The gMFI is calculated for conditions in which a minimum of 5% of the cells were T-bet+. (B) Comparison of T-bet expression (gMFI) by naïve, memory, and GC B cells stimulated in vitro with antigen either attached to PLB or PMS or soluble, in the presence of CpG and IFN-γ (n = 4). Data for T-bet expression (gMFI) by naïve, memory, and GC B cells stimulated in vitro on all stimulation conditions is provided in Supplementary Table 1. Data were analyzed using one-way analysis of variance (ANOVA) with Tukey's adjustment. *P < 0.05; **P < 0.01; ns, not significant. (C) Heat map indicating the percent of T-bet+ cells for each of the conditions in (A).