Figure 2.

HDAd-Derived Cytokines Minimally Enhance the Anti-tumor Effect of HER2.CAR T Cells In Vitro and In Vivo

(A) Schematic structure of HDAd encoding a cytokine expression cassette (HDAdCyto). A549 cells were infected with 100 vps/cell of HDAdCyto. Media were collected 48 hr post-infection, and cytokine levels in the media were measured by ELISA. Data are presented as means ± SD (n = 4). *p < 0.05, **p < 0.001. (B) HER2.CAR T cells expanded with IL-2 were cultured in the presence of 10 ng/mL recombinant cytokines for 30 min, and phosphorylation of STATs was analyzed by flow cytometry. The experiments were repeated with HER2.CAR T cells derived from a second donor with similar results. (C) FaDu or SCC-47 expressing ffLuc cells were infected with 100 vps/cell of HDAdCyto. HER2.CAR T cells were added 24 hr post-infection (effector:target ratio of 1:40). Cells were harvested 120 hr post-co-culture, and viable cancer cells were analyzed by luciferase assay. Data are presented as means ± SD (n = 4). *p < 0.001. (D) SCC-47 cells were transplanted into the right flanks of NSG female mice. A total of 1 × 10⁸ vps of HDAdCyto were injected intra-tumorally. A total of 1 × 10⁶ HER2.CAR T cells were systemically administered 3 days post-injection of HDAds, and tumor volumes were measured at different time points. Data are presented as means ± SD (n = 3).