Figure 6.

Endogenous TRAILR2 or Fas modulate CD40 signalling. (a) NF-κB response of wild-type, Fas KO, TRAILR2 KO and CD40 KO BJAB cell lines induced by the indicated concentrations of Flag-ACRP-CD40L (mega-CD40L). One out of two independent experiments is shown (b). Average of the different clones and replicates of each KO cell line stimulated with 1 μg/ml of mega-CD40L. (c) Ex vivo expression profile of CD40 in marginal zone (MZ) and switched memory (SM) primary human B cells. (d) Expression profile of CD40 in MZ and SM along 6 days of CD40L+IL-21 stimulation. (e and f) Same as c and d, but for Fas expression. (g and h) Same as c and d, but for TRAILR2 expression. MZ B cells are gated as CD27+, IgD+ and SM B cells are gated as CD27+ IgD-. The mean and S.E.M. of six independent experiments is shown. (i) TRAILR2 expression in CD27+ B cells of one representative donor out of four tested after six days of stimulation with CD40L+IL-21. The same gating was used in panel j. (j) CFSE proliferation profile of TRAILR2 low CD27+ (grey histogram) and TRAILR2 high CD27+ (black line) primary human B cells of one representative donor out of four tested after six days of stimulation with CD40L+IL-21. (k) Average proliferative response for the four independent donors analyzed. (l) Flow cytometry analysis of phospho-p65 (RelA) after 5-min stimulation with CD40L. The figure shows five independent donors analyzed in duplicate