Figure 7.

The antitumor activity of ADV^Tα1 is mediated by macrophages and CD8⁺ T cells, and ADV^Tα1 can reprogram the TME toward a more beneficial state for antitumor immunity

(A) Experimental schematic of mice from Figures 7B–1I: 4T1 tumor-bearing WT mice were administered different immunotherapies or vehicle control (PBS) starting on day 10 when the tumor volume reached approximately 50–100 mm³. TILs from the tumor were assessed by flow cytometry on day 24; s.c., subcutaneous; i.t., intra-tumoral.

(B–D) 4T1 tumor-bearing WT mice were administered different immunotherapies or vehicle control (PBS) starting on day 10 when the tumor reached approximately 50–100 mm³ in volume (n = 10 biological replicates). (B) Tumor growth. (C) Body weight. (D) Survival.

(E–I) Representative plots (left), percentages (top), and total cells normalized to g (gram) tumor tissue (bottom) of TILs. (E) “M1-like” macrophages, (F) “M2-like” macrophages, (G) CD25⁺Foxp3⁺ Tregs, and (H) CD3⁺CD8⁺ T cells within the TME of mice. (I) Representative plots and percentages of CD25⁺ CD8⁺ T cells, CD69⁺ CD8⁺ T cells, and IFN-γ⁺ CD8⁺ T cells within the TME of mice (n = 6 biological replicates).

(J–M) 4T1 tumor-bearing mice were administered different immunotherapies and treated with intraperitoneal injections of anti-CSF1R or anti-CD8a; s.c., subcutaneous; i.t., intra-tumoral; i.p, intra-peritoneal. (J) Schematic diagram of the timeline of the antibody depletion experiment in the 4T1 model. (K) Tumor growth. (L) Body weight. (M) Survival (n = 10 biological replicates).

(N–Q) Tumor-bearing NCG mice were intravenously injected with human peripheral blood mononuclear cells on day 1, and then mice were administered different immunotherapies; s.c., subcutaneous; i.t., intra-tumoral; i.v., intravenous. (N) Schematic representation of experimental design and treatment timeline. (O) Tumor growth. (P) Body weight. (Q) Survival (n = 5 biological replicates).

(R–U) The PDX tumor cells were injected into the fourth mammary fat pads of NCG mice, and these tumor-bearing mice were intravenously injected with human peripheral blood mononuclear cells on day 1, followed by different immunotherapies starting on day 10; s.c., subcutaneous; i.t., intra-tumoral; i.v., intravenous. (R) Schematic representation of experimental design and treatment timeline. (S) Tumor growth. (T) Body weight. (U) Survival (n = 5 biological replicates).

The data are shown as the means ± SD. NS, no significant difference; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.