In vivo tumor studies in immunocompetent mice

1.2 million TC1 or TC1-meso cells were subcutaneously inoculated on the right flanks of wild type mice, and after tumors were established (around 200 mm³), a single (unless otherwise indicated) intravenous dose of 10⁸ pfu (100 µl) of viral vectors (VV.luc and VV.CXCL11) was administered. Tumors and mice were then monitored 2–3 times weekly, and tumor volumes were measured using digital calipers. Mice bearing tumors that exceeded 2000 mm³ and sickly mice were euthanized. For combinatorial studies with the Ad.E7 cancer vaccine, TC1-bearing mice were vaccinated subcutaneously on the left flanks (contralateral to the tumor site) with 10⁹ pfu Ad.E7 after tumors were established (approximately 200 mm³). 2 days after vaccination, 10⁸ pfu of VV.luc and VV.CXCL11 were intravenously administered. For combinatorial studies with mesoCAR T cells, 10⁷ murine CAR-expressing T cells were intravenously injected 2 days after intratumoral VV.luc or VV.CXCL11 administration. At endpoint, tumors were harvested from mice and digested as described.^10,29,36,37 Flow cytometry was conducted as described above.