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. 2022 Aug 18;21(9):e13692. doi: 10.1111/acel.13692

FIGURE 1.

FIGURE 1

B cells from older human donors do not have defects in differentiating into plasma cells and proliferating upon stimulation. (a) Gating strategy for plasma cells (CD19+ CD27+ IgD− CD20lo CD38+ IRF4+) 6 days after memory B cells were stimulated with CD40L and IL‐21 or unstimulated (cultured with media). Cells were pre‐gated for live cells and single cells. (b, c) Percentages of plasma cells out of live cells derived from naïve (b) and memory B cells (c) of younger (20–34 years old) and older donors (68–76 years old) after 6 days stimulation with CD40L and IL‐21. Data representative of four independent repeat experiments. (d) Representative flow cytometric histograms showing the cell trace violet stains of live naïve and memory CD19+ B cells from a 24 year old (grey) and 68 year old (blue) donor after 6 days incubation with CD40L and IL‐21 or media (orange). (e) Graph depicting the average number of divisions undergone by proliferating CD19+ naïve and memory B cells after 6 days stimulation with CD40L and IL‐21. (f) Graph showing the percentage of B cells in each division after 6 days stimulation with CD40L and IL‐21. Bar height corresponds to the mean, error bars indicate standard deviation, and each symbol represents values from independent donors. Statistics were calculated using the unpaired Mann–Whitney U test. Data were representative of three independent repeat experiments.