AML Mouse Xenograft Model

Five- to seven-week-old nonobese diabetic (NOD)-Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice were purchased from the Jackson Laboratory and maintained at the Baylor College of Medicine Animal Facility. All procedures were done in compliance with the Institutional Animal Care and Usage Committee of Baylor College of Medicine. Mice were sublethally irradiated (116–200 cGy) and injected intravenously 24 hr later with 1 × 10⁶ KG-1a-FFluc cells followed by a single injection of 2 × 10⁶ of CD7 CAR T cells 5 or 8 days later. Tumor burden was monitored using an IVIS Imaging system (Caliper Life Sciences) by recording bioluminescence from mice injected with 150 mg/kg of D-luciferin intraperitoneally at indicated time points. Living Image software (PerkinElmer) was used to visualize and calculate total luminescence. For analysis of tumor cells in peripheral blood, 100 μL of blood was collected by tail-vein bleeding. After red blood cell lysis, cells were incubated with anti-human CD45 and CD3, and CD7 antibodies for subsequent flow cytometric analysis.