Induction of plasma cells from CD27− naive B cells.
Highly purified adult CD27− B cells were cultured with SAC (0.01%) + IL-2 (50 U/mL), or IL-10 (50 ng/mL) + anti-CD40 mAb (1 μg/mL), SAC + IL-2 + IL-10 + anti-CD40 mAb in conjunction with cross-linking by CD32T (40%) at a final cell density of 0.5 × 105 per well in 96-well round-bottom plates for 8 to 10 days. Alternatively, the cells were cultured with IL-4 (50 ng/mL) + anti-CD40 mAb in conjunction with cross-linking by CD32T (40%) at a final cell density of 1 × 105 per well for 12 to 14 days. The cells were then stained with anti-CD38–FITC and anti-CD20–PE. The antibody-coated cells were gated on living cells by cell size and granularity, and dead cells were removed by propidium iodide (PI) staining and then counted by means of flow cytometric analysis. Expression of CD38 and CD20 on B cells are shown with a log scale. Data are representative of the results of 2 experiments using different donors. The same results were obtained when CB B cells were cultured with above-mentioned stimuli.
