Fig. 4: Impact of PDCNP-Ab on the TME.

PDCNP-Ab (125 μg·Ca/kg, loaded with 0.5 wt% PMA) were administered i.t. For comparison, equivalent doses of DCNP-Ab, calcium salt, ION/PMA (6.7 nM/kg and 250 ng/kg, respectively), and PBS were injected. a Timeline of nanoparticle injections and flow cytometry analysis; experiments were conducted in B16F10-OVA tumor-bearing mice. Created in BioRender. Li, J. (2024) https://BioRender.com/i82v387. b Profiles of tumor-infiltrating T cells, including T cells (CD3⁺CD8⁺), effector T cells (CD3⁺CD8⁺IFNγ⁺), proliferating T cells (CD3⁺CD8⁺Ki67⁺), and Tregs (CD3⁺CD4⁺FoxP3⁺) (n = 5 mice). T cell expansion (c) and activation (d), investigated ex vivo by co-culturing splenocytes isolated from different treatment groups with B16F10-OVA cells (n = 5 mice). Influence on T cell exhaustion. e Frequencies of Eomes⁺, PD1⁺Eomes⁺, and T-bet⁺ cells in CTLs as well as T-bet⁺/Eomes⁺ ratios among CTLs (n = 5 mice). f MFI of LAG3 and TIM3 in PD-1⁺ CTLs (n = 5 mice). The experimental scheme is shown in Supplementary Fig. 24a. g Changes of naïve, T_CM, and T_EM populations among spleen CTLs, measured one week after treatment with PDCNP-Ab or PBS (n = 3 biologically independent samples). h Therapeutic treatment regimen. The experiments were conducted in B16F10-OVA tumor-bearing mice. Irradiation (RT) was applied to tumors on Day 1. PDCNP-Ab and controls were administered i.t. on Day 2, 5, and 8. Animals were euthanized on Day 13 for flow cytometry analysis. Created in BioRender. Li, J. (2024) https://BioRender.com/m87w815. i Profiles of tumor-infiltrating T cells, including T cells (CD3⁺CD8⁺), effector T cells (CD3⁺CD8⁺IFNγ⁺), and Tregs (CD3⁺CD4⁺FoxP3⁺). In addition, splenocytes from the treatment groups were co-cultured ex vivo with B16F10-OVA cells (n = 5 mice). j Levels of cytokines, including IL-2, TNF-α, and IL-10, in tumor tissues, analyzed by ELISA (n = 3 biologically independent samples). Data represent mean ± SD. Statistical difference was evaluated using an unpaired Student’s t-test (one-tailed) in (g), and a one-way ANOVA test in (b, d–f, i, j). Source data underlying (b–f) and (g–j) are provided in a Source Data file.