Figure 11. Effect of TCDD on CD40L-induced immediate MAPK and Akt activation in human B cells at 30 min.
Freshly isolated human B cells (the same human donor to Figure 13) (1 × 106/ml) were equilibrated at 37°C for 3–4 h, treated with 3, 10 and 30 nM of TCDD, and/or vehicle (VH), or remained untreated (NA) as a vehicle control and then stimulated with recombinant human CD40L (200 nM), human IL-2 (10 U/ml), IL-6 (100 U/ml), and IL-10 (20 ng/ml) for 30 min. Cells were fixed with 1.5% paraformaldehyde and permeabilized with ice-cold 99% methanol. Cells were then assessed by multiparametric phosflow for intracellular expression of Akt and p38. Three replicates were included per group. The percentages of cells that expressed phospho-ERK, phospho-p38, and phospho-Akt were obtained using the Boolean gating technique of the FlowJo software. *, p < 0.05, compared to the VH-treated group.