Figure 5.

IN administration of antigen with NE adjuvants after IM mRNA priming effectively pulls antigen-specific immunity to mucosal sites

Single cell suspension were isolated from the spleen (A–D), cLNs (E–H), and lungs (I–L) of mice given prime/boost immunizations with the indicated adjuvant/antigen regimens. Mice were given 3 μg mRNA IM, and 20 μg of S protein IN in either PBS, NE, or NE/IVT. Cells were stimulated with 25 μg/mL of S protein, and antigen-specific cytokine responses were quantified in CD4⁺ T cells by intracellular cytokine staining and FACS analysis. (n = 4–5/group with data represented as mean ± SEM; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by one-way ANOVA with Tukey post hoc test shown only for select groups (full statistical analysis is shown in Table S1).