Relative Contribution of Th1 and Th17 Cells in Adaptive Immunity to Bordetella pertussis: Towards the Rational Design of an Improved Acellular Pertussis Vaccine (original) (raw)
Figure 1
Th17 and Th1 cells mediate host immunity to B. pertussis in the respiratory tract of naive mice.
(A–C) Naive C57BL/6 mice were exposed to an aerosol infection with B. pertussis and groups of 4 mice were sacrificed at the indicated time points. (A) Lung mononuclear cells were stimulated with heat-killed B. pertussis and after 3 days of culture IL-17A was quantified in supernatants by ELISA. (B–C) lung mononuclear cells were incubated with brefeldin-A for 1 h and intracellular cytokine staining for IL-17A, together with surface staining for CD4 was performed, followed by FACS analysis. Results are expressed as mean frequencies of IL-17A+CD4+ cells (B), with sample FACS plots (C) (D–E) C57BL/6 WT and IL-17A−/− mice were aerosol challenged with B. pertussis and groups of 4 mice were sacrificed at the indicated time points. CFU counts were performed on lung homogenates (D) ** p<0.01 IL-17A−/− versus WT. Neutrophil recruitment was determined by FACS analysis on lung lavage (E). (F) Spleen cells from IFN-γ−/− or WT mice that had cleared a respiratory infection with B. pertussis were stimulated in vitro with killed B. pertussis and IL-12 (Th1) or IL-1β and IL-23 (Th17) respectively. After 4 days of culture surviving cells were harvested and _B. pertussis_-specific Th1, Th17 or both (10×106) were transferred to naive mice, which were aerosol challenged with live B. pertussis 24 hours later. Naive mice that did not receive a cell transfer and mice injected with T cells from a naive mouse were used as controls. The course of infection was followed by performing CFU counts on the lungs at intervals after challenge. +p<0.05, +++ p<0.001 Th1+Th17 versus control; ** p<0.01, *** p<0.001 Th17 versus control. Results (except panel C) are mean values for 4 mice per group at each time point and each panel is representative of either 3 to 4 independent experiments.