Figure 2.

Recombinant IFN‐α enhances SEA/aCD40‐induced B‐cell IL‐10 production. (A–B) B cells were isolated from the spleen of naϊve mice and stimulated in vitro with SEA (20 µg/mL), anti‐CD40 (0.5 µg/mL), and IFN‐α (10³–10⁵  U/mL) as indicated. After 3 days of culture, supernatants were analyzed for IL‐10 and IL‐6 concentration by ELISA (A), and the percentage of IL‐10⁺ B cells assessed by flow cytometry (B). (C–D) Splenic marginal zone and follicular B cells from naïve mice were sorted using flow cytometry and cultured for 2 days in the presence of SEA (20 µg/mL), anti‐CD40 (0.5 µg/mL), and IFN‐α (10⁴/mL). IL‐6 and IL‐10 production as measured by CBA (C) and frequency of IL‐10⁺ cells in each respective subset as determined by flow cytometry (D). Data are pooled from four (A) or three (B–D) experiments; each data point is the mean of two to four technical replicates. Data are presented as mean +SEM. Significant differences are indicated by *p < 0.05, **p < 0.01, ***p < 0.001 and determined by one‐way ANOVA followed by Dunnett's multiple comparisons test. ^# p < 0.05, ^## p < 0.01, ^### p < 0.001 indicate significant difference relative to medium only control.