Figure 2.

DC101 normalized the tumor vasculature and increased the number of intratumoral CD8⁺ T cells. H22 cells were subcutaneously inoculated into Balb/c mice. When the tumor volume reached ~170 mm³, DC101 10 mg/kg was administered through i.p. injection on days 2, 5, and 8. (A1) Immunofluorescent confocal images of the tumors showed blood vessel pericyte coverage in the PBS and DC101 groups on day 10. CD31⁺ endothelial cells, red; α-SMA⁺ pericytes, green; nuclei, blue. (A2) Immunofluorescent images for the Hoechst 33342 (Hoechst) tumor perfusion assay in the PBS and DC101 groups on day 10. Hoechst 33342⁺ cells, blue; nuclei, green. (A3) Immunofluorescent images of the tumors showing FITC-lectin (lectin) tumor blood vessel perfusion in the PBS and DC101 groups on day 10. CD31⁺ endothelial cells, red; FITC-lectin-perfused vessels, green; nuclei, blue. (A4) Immunofluorescent images showing the level of hypoxia in the PBS and DC101 groups on day 10. Pimonidazole⁺ cells, red; nuclei, blue. (B1) Quantification of pericyte coverage determined through α-SMA⁺CD31⁺/CD31⁺ staining using ZEN software (Carl Zeiss, Germany) (n = 3). (B2) Quantification of tumor perfusion determined through the Hoechst 33342⁺ area/field using the Image J software (n = 3). (B3) Quantification of perfused functional tumor blood vessels determined by lectin⁺CD31⁺/CD31⁺ staining using ZEN software (Carl Zeiss, Germany) (n = 3). (B4) Quantification of tumor hypoxia determined by pimonidazole⁺ area/tumor area using ZEN software (Carl Zeiss, Germany) (n = 3). (C) Representative flow cytometry plots for intratumoral CD4⁺ T (CD3⁺CD4⁺) cells and CD8⁺ T (CD3⁺CD8⁺) cells in the PBS and DC101 groups on day 10. (D, E) Quantification of intratumoral CD4⁺ T cells (D, n = 3) and CD8⁺ T cells (E, n = 3) detected by flow cytometry. Data are presented as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001).