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. 2024 Feb 22;18(9):6845–6862. doi: 10.1021/acsnano.3c04471

Figure 5.

Figure 5

Nanoparticle vaccine enhances dendritic cell activation and cross-presentation, activates antigen-specific CD8+ T cells, and provides therapeutic efficacy in the EG7.OVA model. (A) Flow cytometric quantification of mean fluorescence intensity (MFI) of CD86 expression by BMDCs treated with indicated formulations (mean ± SD; n = 2; **P < 0.01; one-way ANOVA with Tukey’s multiple comparisons). (B) Flow cytometric quantification of mean fluorescence intensity (MFI) or percentage of SIINFEKL-H2Kb positive BMDCs treated with indicated formulations and stained with PE-labeled antibody against SIINFEKL/H-2Kb (mean ± SD; n = 3; ****P < 0.0001; one-way ANOVA with Tukey’s multiple comparisons). (C) Vaccination and downstream analysis scheme. (D) Quantification of the frequency of SIINFEKL-specific CD8+ T cells in spleen after vaccination using peptide/MHC tetramer staining (mean ± SD; n = 10 mice/group; ****P < 0.0001; one-way ANOVA with Tukey’s multiple comparisons). (E) Quantification of the frequency of SIINFEKL-specific CD8+ T cells in spleen after vaccination using peptide/MHC tetramer staining comparing wild-type mice to Batf3–/– mice (mean ± SD; n = 5–8 mice/group; ***P < 0.001, ****P < 0.0001; two-way ANOVA with Tukey’s multiple comparisons). (F) Tumor challenge and therapeutic vaccination scheme. (G) Average EG7.OVA tumor growth in response to indicated formulation (mean ± SEM; n = 8–10 mice/group). (H) Average tumor volume on day 20 after tumor inoculation (mean ± SEM; n = 8–10 mice/group; **P < 0.01, ***P < 0.001; one-way ANOVA with Tukey’s multiple comparisons). (I) Kaplan–Meier survival curve using 2000 mm3 tumor volume as the end point (n = 8–10 mice/group; statistical significance between indicated treatment versus PBS and NP-Pep/cGAMP; *P < 0.05, **P < 0.01; Mantel–Cox log-rank test). (J) Representative flow cytometry plots and quantification of IFNγ+TNFα+ CD8+ T cells after ex vivo restimulation of splenocytes with SIINFEKL peptide (mean ± SD; n = 8 mice/group; ****P < 0.0001; unpaired t test). (K) Percentage of central and effector memory antigen-specific CD8+ T cells (TCM and TEM) in spleen after vaccination (mean ± SD; n = 8 mice/group; ****P < 0.0001; unpaired t test).