Isolation and Activation of T Cells

Peripheral blood lymphocytes (PBLs) from independent normal human donors were isolated from purchased apheresis products (Key Biologics, Memphis, TN) using Ficoll-Hypaque (Sigma-Aldrich, St. Louis, MO) density gradients^{40 (link),43 (link)}. T cells were activated by stimulating PBLs from an individual donor with 50 ng/mL anti-CD3 mAb (Miltenyi Biotec, Bergisch Gladbach, Germany), 300 IU/mL recombinant human IL-2 (rhIL-2; Novartis Pharmaceuticals, East Hanover, NJ), and 100 ng/mL recombinant human IL-15 (rhIL-15; Biological Resources Branch, National Cancer Institute, Bethesda, MD) for three days prior to transduction^{43 (link)}. Alternatively, PBLs from the same donor were cultured with 20 ng/mL recombinant human IL-7 (rhIL-7; Biological Resources Branch, National Cancer Institute) for seven days prior to transduction.