Fig. 3

Effects of anti-CD40 antibodies KPL-404 and G28-5 on B cell proliferation. PBMCs were labeled with a cell proliferation tracker dye (Tag-it Violet) and cultured for 5 days in the presence of 10 μg/ml IgG4 isotype control Ab or anti-CD40 Abs—KPL-404 and G28-5. Cells were left untreated (media control) or stimulated with anti-CD3/CD28 cross-linking reagent ImmunoCult (IC). Levels of CD19⁺ B cell proliferation were measured based on Tag-it Violet dilution. a Flow cytometry data from one representative experiment. Gates depict non-proliferating cells (gate P0) or Tag-it Violet dim (proliferating) cells (gates P1-P2). b Cumulative data from 3 individual donors. B cell proliferation is presented as division Index (total number of divisions/total number of cells) for each sample. The bars represent mean and standard deviation (SD). *p < 0.05, determined by two-way ANOVA with multiple comparisons test