Deletion of AMPK decreases xenogeneic GVHD severity without affecting antileukemia potential. (A-C) A total of 6 × 106 mock or CRISPR-treated human T cells were transplanted with 1 × 106 autologous non–T cells into irradiated NSG recipients, and recipients followed for survival (A) and clinical score (B) to 12 weeks posttransplant. Increased clinical scores were driven by significant skin manifestations, exaggerated fur ruffling, and dramatically hunched posture (representative photographs shown in panel C). To assess antileukemic potential, mock vs CRISPR–treated human T cells were plated at an effector-to-target ratio of 2:1 with GFP+ Molm13 leukemia cells and cytotoxicity measured using an Incucyte analyzer and the loss of Green integrated (G.I.) intensity over time (D). A similar approach was used to measure cytotoxicity of mock vs CRISPR–treated, CD19–targeting CAR T cells placed into Incucyte incubator with GFP+ Nalm6 leukemia cells (E). Ten to 11 mice per group in panels A-B, with clinical manifestations shown for 2 representative animals in panel C. Experiments in panels D-E are representative of similar results using T cells from 2 independent donors.
