Cell proliferation and cytokine generation. (A) Enriched B cells were stained with CFSE (1 μM) and stimulated with F(ab′)2 specific for anti-IgM (10 μg/mL), IL-4 (50 ng/mL), and CD40 ligand (10 μg/mL) for 5 days. Percent cells having undergone at least one cellular division, (numbers above the lines), assessed by CFSE dilution. Gray solid peak is an unstimulated cell. (B) EBV-transformed B cells from 3 normal and 2 different batches of patient were counted for 7 days. (C) PBMCs were stained with CFSE (1 μM) and stimulated Dynabeads T-cell activator CD3/CD28 for 3 days. PHA, phytohemagglutinin. (D) and (E) Enriched B cells were stimulated as indicated. After 48 hours, cell-free supernatants were harvested and cytokines measured as described in the Cytokine measurement section in the Materials and methods. Data from panels A and C are representative of 3 independent experiments. Data from panels D and E were represented as means ± SE of n = 3 separate experiments.
