Whole blood was obtained from de-identified healthy donors at the DeGowin Blood Center at the University of Iowa Hospitals and Clinics. The recruitment protocol and written informed consent document were approved by the Institutional Review Board for the University of Iowa. PBMCs were isolated from leukocyte reduction system (LRS) cones using Hypaque-Ficoll density-gradient separation as previously described (33 (link)). T cells were then expanded and activated with anti-CD3- and anti-CD28-coated beads (Invitrogen) and human IL-2 for 5 days. Cells were then re-suspended in fresh medium without stimulatory beads or IL-2 for 24 hours. Activated T cells were resuspended in serum-free RPMI 1640 before GML treatment. GML was solubilized at room temperature in 95% ethanol and diluted into the appropriate working concentration. We added 95% ethanol as a comparative vehicle control at final concentrations that did not exceed 0.5%. Activated T cells were treated with ethanol or various doses of GML in serum-free medium for various assays.