Establishment of LUSC xenografts and treatment

The animal studies were approved by the ethics committee of the Peking University Cancer Hospital & Institute. NOD-SCID and C57BL/6N mice were obtained from Vitalriver. Human LUSC cell line H520 or PDX tumors were subcutaneously injected into NOD-SCID mice, and the mouse lung cancer cell line LEWIS was subcutaneously inoculated into C57BL/6N mice for xenograft establishment. In addition, the orthotopic model was established in the lung of NOD-SCID mice using H520 cells with the forced expression of MECOM and firefly luciferase. When the tumor volume reached 50–200 mm, the mice were used for further experiments.

NOD-SCID mice with H520 tumors and C57BL/6N mice with LEWIS tumors were treated with ADV (1.0 × 10¹⁰ viral particle [VP]) and CFS and HF protein complex (1.0 × 10⁻⁷ pmol/VP) via the tail vein to analyze the distribution, specificity, and immunogenicity of ADV. To determine the therapeutic potential of ADV (0.7 × 10¹⁰ VP) and protein complex (1.0 × 10⁻⁷ pmol/VP), 2–4 rounds (12–14 days intervals) of the complex were administered by intratumoral or intravenous injection.