Fig. 2.

GM-CSF and FLT3L BMDCs secrete different cytokine profiles and activate CD8⁺ T cells in response to S1 protein with combination adjuvants. A) t-SNE plots of GM-CSF and FLT3L BMDCs with labeled clusters of APC subsets. Macrophages/Mo-DCs are CD64⁺ F4/80⁻, Conventional DCs are CD11c⁺ MHCII⁺ CD64^lo F4/80^lo, Monocytes are Ly6C^hi CD11c⁻, Neutrophils are Ly6G^hi Ly6C⁺, plasmacytoid DCs are B220⁺ Ly6C⁺ CD11c⁺ MHCII^lo. B–G) Cytokine concentrations (pg/mL) of IL-1β, IL-27, IL-12p70, IFN-β, and IFN-λ3 in supernatants of BMDC culture after incubation with adjuvanted NPs for 24 h. H–I) Percentage of live CD3⁺CD4⁺ T cells or CD3⁺CD8⁺ T cells proliferating in presence of GM-CSF BMDCs, gated on diminished CFSE signal (Fig. S1). “No Adjuvant” condition is blank NPs. J, K) Percentage of live CD3⁺CD4⁺ T cells or CD3⁺CD8⁺ T cells proliferating in presence of FLT3L BMDCs. *p < 0.05. **p < 0.01, ***p < 0.001, ****p < 0.0001 based on a One-Way ANOVA and Tukey Test for multiple comparison.