Fig. 3: Aerosol-delivered AIBP vaccine induces respiratory TH1 and TH17 TRM cells and confers protection against B. pertussis infection of the lungs and nasal cavity.
Female 6–8-week-old C57BL/6 mice were immunized by aerosol administration of the AIBP vaccine (once or twice at 0 week and 4 weeks), an aP vaccine (i.m., twice, 0 week and 4 weeks; 1/50 human dose) or PBS. The mice were aerosol challenged from a culture at 1 × 109 CFU ml−1 of live B. pertussis at week 6. On the day of but before the challenge with live B. pertussis, mice were injected intravenously with anti-CD45 antibody 10 min before euthanasia (to identify tissue-resident cells), and lung or nasal tissue cells were stained with antibodies specific for TRM cells, or cells were stimulated with HKBP, anti-CD28 and anti-CD49d (both 1 μg ml−1) for 16 h, followed by Brefeldin A (5 μg ml−1) for the final 4 h of culture before intracellular cytokine staining (ICS) and flow cytometric analysis. a, Mean absolute number of CD4 TRM cells (CD45iv− CD4+CD44+CD62L−CD69+CD103+/−) quantified in lung and nasal tissue by flow cytometric analysis. Data are mean ± s.e.m. (n = 5). b, Number of B. pertussis-specific IFNγ- or IL-17-secreting CD45iv−CD4 TRM cells in lung and nasal tissues determined using ICS and flow cytometry. Data in a and b are presented as the mean ± s.e.m. of biological replicates shown as individual symbols (n = 5). c, Before the B. pertussis challenge, concentrations of FHA-specific IgA in nasal tissue homogenates and FHA-specific IgG1 and IgG2c in serum were quantified by ELISA. Data are presented as the mean ± s.e.m. of biological replicates (n = 5). d, Live bacterial loads in lung and nasal tissue were quantified by CFU counts at 2 h and 7 days, 14 days and 21 days after the live B. pertussis challenge. Data are presented as the mean ± s.e.m. of biological replicates (n = 5). Data were analysed by one-way ANOVA followed by Tukey’s test for multiple comparisons. P values are shown above relevant datasets.
