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. 2017 Aug 3;8:914. doi: 10.3389/fimmu.2017.00914

Figure 4.

Figure 4

The number of CD19+CD24hiCD27+ B cells decreased, and their function was impaired in Crohn’s disease (CD) patients. (A) Freshly derived CD19+ B cells from CD patients and healthy controls (HCs) (N = 20 in both cases) were analyzed for interleukin-10 (IL-10) expression by FACS. The percentage of IL-10-secreting cells is shown (NS: no significant difference). (B) PBMCs from CD patients and HCs (N = 15) were analyzed for different B cell subsets using CD24 and CD27 markers, and the CD24 and CD27 antibodies isotype control are showed. The bars represent the mean ± SEM (right panel). (C) CD19+ B cells from HCs and CD patients (N = 15) were cultured with 100 nM CpG oligonucleotides for 48 h with the stimulation or inhibition cocktail added during the final 5 h of culture, and the percentages of IL-10+ B cells within the indicated B cell subsets were determined by FACS. The data are shown as the mean ± SEM. (D) Purified B cell subsets from the blood of HCs and CD patients were stimulated with 100 nM CpG oligonucleotides for 24 h and cultured with CD14+ monocytes isolated from HCs for 24 h; then, the cytoplasmic TNFα expression in CD14+ monocytes was assessed after 4 h of stimulation with 100 µg/mL LPS. The data are shown as the mean ± SEM (N = 5). (E) Purified B cell subsets from the blood of HCs and CD patients were stimulated with 100 nM CpG oligonucleotides for 24 h and cultured with CD3 mAb-stimulated CD4+CD25− T cells isolated from HCs for 48 h. IFNγ expression in the CD4 T cells was analyzed by FACS. The data are shown as the mean ± SEM (N = 3). *P < 0.05; **P < 0.01; NS, no difference.