Extended Data Fig. 5: Aerosol-delivered AIBP vaccine induces B. pertussis-specific T_H1- and T_H17-type T_RM cells in the lungs and nasal tissue.

Female 6-8 week old C57BL/6 mice were immunized by aerosol administration of AIBP vaccine (once or twice at 0 and 4 weeks), i.m. immunization with an aP vaccine (twice, 0 and 4 weeks;1/50 of the human dose) or PBS. Mice were aerosol challenged from a culture at 1×10⁹ CFU/mL of live B. pertussis at week 6. On the day of but prior to challenge with live B. pertussis, mice were injected i.v. with anti-CD45 antibody 10 min prior to euthanasia (to identify tissue-resident cells), and lung or nasal tissue cells were stained with antibodies specific for T_RM cells, or cells were stimulated with HKBP, anti-CD28 and anti-CD49d (both 1 μg/mL) for 16 h, followed by Brefeldin A (5 μg/ml) for the final 4 h of culture prior to intracellular cytokine staining (ICS) and flow cytometric analysis. Representative flow plots of CD69 and CD103 expression on CD45iv⁻CD4⁺CD44⁺CD62L⁻ T cells in lung tissue (a) or nasal tissue (b). Representative flow plots of B. pertussis-specific IFNγ- or IL-17-secreting CD45iv⁻CD4⁺ T_RM cells in lung tissue (c) or nasal tissue (d).

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