Tumor Xenograft Model for Evaluating Immunotherapy

Huh7 (1 × 10⁷/200 μL) cells were injected subcutaneously into the right axilla of BALB/c nude mice (Vital River Laboratories, Beijing, China). When the tumors grew to average diameter of 5 mm, the animals were randomized into 5 groups and received intravenous injection of either PBS, CIK cells, KGHV500, CIK + KGHV500, or CIK + KGHV400, respectively. Tumor size was measured with Vernier caliper every three days. Tumor volume was calculated according to the formula: V= (length × width²) ÷ 2 16 (link). When tumors in PBS group became ulcerated, mice were sacrificed. The dissected tumors were embedded with paraffin for immunohistochemical analysis. The anti-FLAG tag antibody (Abnova, MAB9744, China) and anti-human adenovirus hexon mouse mAb (Novus, NBP211638, USA) were used to detect KGHV500 adenovirus.