Fig. 7: Intrapulmonary administration with Advax elicits innate and adaptive cell activation that is enhanced by presence of CysVac2 antigen.

C57BL/6 mice were vaccinated i.t. with PBS, Advax-FITC or CysVac2/Advax-FITC and lungs and mediastinal lymph nodes were harvested for mass cytometric analysis 7 days later. Samples were treated with protein transport inhibitor cocktail (eBioscience) prior to fixation for intracellular staining. a–i Bar graphs depict the cell number of subsets ±SEM identified via CyTOF in the lungs of vaccinated animals. j Heat maps of geometric mean fluorescence intensity (MFI) expression of activation markers of (A) PBS, (B) Advax or (C) CysVac2/Advax vaccinated by lung cell subsets were generated using Prism 8 software, showing relative upregulation or downregulation in relation to PBS-treated group. Mean values represent n = 3–4 mice per group and are representative of two independent experiments. Significance of differences between groups was determined by ANOVA with post-hoc Tukey’s multiple comparison test (*p < 0.033, **p < 0.0021, ***p < 0.0002, ****p < 0.0001).