Bordetella pertussis Commits Human Dendritic Cells to Promote a Th1/Th17 Response through the Activity of Adenylate Cyclase Toxin and MAPK-Pathways (original) (raw)
Figure 4
Analysis of MyD88 -dependent pathway induction.
A MDDC were either untreated or treated with BpWT or BpCyaA−. Phosphorylation of p38, ERK1/2, SAP/JNK and IκB-α was determined at the indicated time-points by Western blot. A single gel was run and blotted to detect phosphorylated proteins and β tubulin to normalize the results. Data are from one representative out of four independent experiments performed with MDDC obtained from different donors. B MDDC were treated as in panel A, either in the absence or presence of p38 inhibitor (SB203580), ERK1/2 inhibitor (PD98059) or PI3K inhibitor (LY294002) for 48 h. Results of seven independent experiments performed with MDDC obtained from different donors are expressed as the percent of change of maturation markers (CD80, CD83, CD38) with respect to the corresponding stimulus in the absence of inhibitors. Mean ± SE of marker expression in MDDC not treated with inhibitors was for BpWT: CD80 (MFI) = 48±6; CD83 (%) = 23±6; CD38 (MFI) = 18±4; for BpCyaA−: CD80 (MFI) = 72±11; CD83 (%) = 36±7; CD38 (MFI) = 46±5. * p<0.05 vs control, calculated from the raw data. C MDDC were treated as in panels A and B. Results of ten (IL-12p70), eight (IL-23 and IL-10) and 5 (IL-1β) independent experiments performed with MDDC obtained from different donors, measured by ELISA, are expressed as the percent of change of cytokines with respect to the corresponding stimulus in the absence of inhibitors. Mean ± SE of cytokine production (pg/ml) in MDDC not treated with inhibitors was for BpWT: IL-12p70 = 0±0; IL-23 = 310±70; IL-1β = 330±85; IL-10 = 2034±605; for BpCyaA−: IL-12p70 = 374±43; IL-23 = 672±140; IL-1β = 600±87; IL-10 = 2477±593. * p<0.05 vs control, calculated from the raw data.