Fig 1.
Regulatory B10 cells are augmented and the level of IL-10 expression is decreased in the spleen of AD mice.
(A) Representative dot-plots showing the percentage of CD5+CD19+CD1dhi B10 cells in control (n = 10) and AD mice (n = 10); (B) Bar graph depicting the percentage of CD5+CD19+CD1dhiB10 cells in control and AD mice as analyzed by flow cytometry. The percentage of CD5+CD19+CD1dhi B10 cells was significantly higher in the AD group than that observed in the control. Data are expressed as mean ± SEM.*P<0.05, asanalyzed by one-way ANOVA, followed by Tukey multiple-comparison test; (B) The level ofIL-10 mRNA expression decreased in the spleen of AD mice (n = 10) compared to that of control (n = 10). Data are expressed as mean ± SEM. *P < 0.05, as analyzed by one-way ANOVA, followed by Tukey multiple-comparison test.
Fig 2.
Comparison of IL-10 production in CD19+ B cells of AD mice and control.
The B10 cells in the spleen of control (n = 10) or AD mice (n = 10) were in vitro stimulated for 5h or 48h withLPS(10μg/mL),PMA (50ng/mL), iono(500 ng/mL),monensin (2 mM), and CD40(1μg/mL). The number of CD19+IL-10+B cells wasmeasured by flow cytometry by using intracellular cytokine staining. (A) Representative comparative phenotype of CD19+IL-10+B cells from a control and AD mouse at 5 h and 48 h of stimulation; (B) The mean (±SEM) frequency of CD19+ B cells in PBMCs of spleen between controland AD groups after cultured for 5 h; (C) The mean (± SEM) frequency of B10pro+B10 cells in PBMCs of spleen between control and AD groups after culturing for 48 h; Data are expressed as mean ± SEM. Data are expressed as mean ± SEM. **P < 0.01, as analyzed by one-way ANOVA, followed by Tukey multiple-comparison test.
Fig 3.
Influence of CD5+CD19+CD1dhi B10 cells on IgE production.
B10 cells were sorted from the spleen of AD mice (n = 5) and control mice (n = 5) by flow cytometry. The PBMCs from normal mice and B10 cells from control or AD mice were cultured with LPS (10 μg/mL), PMA (50 ng/mL), and iono (500 ng/mL) for 72h,then the IgElevels were determined in the supernatants by ELISA.PBMCs without B10 cell wereused as a negative control. No differences in IgE levels were observed in the AD and negative control groups, and both groups showed significantly higher IgElevel than that observed in the control group. Data are expressed as mean ± SEM. **P < 0.01, as analyzed by one-way ANOVA, followed by Tukey multiple-comparison test.