Fig. 6: Improved antitumor effects and antitumor immunity of anti-PD-1 antibody by R848@M2pep-MPs_AFP in orthotopic Hepa1-6 tumor-bearing mice.

a Schematic schedule for the antitumor experiment of combination of R848@M2pep-MPs_AFP and anti-PD-1 antibody in orthotopic Hepa1-6 tumor-bearing mice. b, c Tumor images (b) and tumor weight (c) of orthotopic Hepa1-6 tumor-bearing mice after treatment with R848@M2pep-MPs_AFP in the presence or absence of anti-PD-1 antibody at the anti-PD-1 antibody dosage of 5 mg kg⁻¹ and R848 dosage of 0.5 mg kg⁻¹ indicated in (a). Data are presented as means ± s.d. for (c). (n = 5 mice per group; one-way ANOVA followed by Tukey’s HSD post-hoc test). d Kaplan–Meier survival plot of orthotopic Hepa1-6 tumor-bearing mice after treatment indicated in (a). (n = 8 mice per group). e–l The numbers of CD8⁺ T cells (e), CD8⁺Ki67⁺ T cells (f), CD8⁺CD69⁺ T cells (g), CD8⁺IFNγ⁺ T cells (h), CD8⁺GzmB⁺ T cells (i), CD8⁺PD-1⁺TCF-1⁺ T cells (j), CD8⁺PD-1⁺TCF-1^- T cells (k) and CD8⁺PD-1⁺TCF-1^-GzmB⁺ T cells (l) in tumor tissues of orthotopic Hepa1-6 tumor-bearing mice after treatment indicated in (a). Data are presented as means ± s.d. (n = 5 mice per group; one-way ANOVA followed by Tukey’s HSD post-hoc test). m Schematic schedule for ELISPOT and T cell cytotoxic assay. n, o Images (n) and numbers (o) of IFNγ immune spots from OVA_257-264- or AFP₂₁₂-restimulated splenocytes isolated from orthotopic Hepa1-6 tumor bearing mice after treatment indicated in (m) by the ELISPOT assay. Data are presented as means ± s.d. (n = 5 biological independent samples; two-way ANOVA followed by Bonferroni’s multiple comparisons post-test). p Cytotoxicity of T cells against Hepa1-6 cells when T cells isolated from OVA_257-264- or AFP₂₁₂-restimulated splenocytes were incubated with Hepa1-6 cells at the effector/target ratio of 10:1 for 6 h as indicated in (m) by LDH assay. Data are presented as means ± s.d. (n = 5 biological independent samples; two-way ANOVA followed by Bonferroni’s multiple comparisons post-test). Source data are provided as a Source Data file.