Figure 3.

Effect of recombinant adenovirus Ad5sPVR on antitumor immune activation and lymphocyte infiltration

(A) Schematic diagram of the experimental setup for adenovirus therapy in H22-challenged mice. On day 0, male C57BL/6 mice were injected intraperitoneally with 5 × 10⁶ H22 cells, and the mice were randomly divided into three groups: saline group, Ad5con group, and Ad5sPVR group. On days 8, 12, and 16, mice in the saline group, Ad5con group, and Ad5sPVR group were injected intraperitoneally with saline, the control virus Ad5con (5 × 10⁸ PFU), and Ad5sPVR (5 × 10⁸ PFU), respectively. On days 12 and 16, before intraperitoneal injection of saline or virus, ascites were collected. (B) Virus replication was measured in the ascites of mice infected with recombinant adenovirus. On day 16, before intraperitoneal injection of saline or virus, ascites were collected and the viral copy number was determined by RT-PCR. (C) On day 16, before intraperitoneal injection of saline or virus, ascites were collected and the concentration of sPVR in ascites was determined by ELISA. (D) On day 16, before intraperitoneal injection of saline or virus, ascites were collected, and CD4⁺ T cell, CD8⁺ T cell, and NK cell frequencies in ascites were determined by flow cytometry. (E) On day 16, before intraperitoneal injection of saline or virus, ascites were collected, and immune activity in the TME was detected using a mouse IFN-γ ELISpot assay, and the number of IFN-γ spots in each group was calculated. The data are shown as the means ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. i.p., intraperitoneal; s.c., subcutaneous; i.t., intratumoral.