Naive cells display greater in vitro expansion than memory cells. Cryopreserved leukapheresis samples were thawed and rested overnight. CD8⁺ T-cell subsets were isolated then stimulated with allogeneic feeders, anti-CD3, and IL-2 (A) or stimulated with CD3/CD28 beads and IL-2 (B-C), then transduced to express the 1G4-α-95:LY mutant TCR 48 hours after stimulation. (A) Fold expansion of each culture was determined 10-16 days following stimulation. The fold expansion of CD8⁺ T-cell subsets from 12 patients is displayed (*P < .01, **P < .001). (B) Cells were pulsed with H³ -TdR 48 hours after stimulation, and incorporation as determined 16 hours later is shown. N = 6. (*P < .01, **P < .001). (C) Cell division was determined by CFSE dilution 3 days after stimulation (shaded histograms). The frequency of cells within each gate is indicated. Open histograms are undivided controls. The x-axis is on a log scale and the y-axis is on a linear scale. Data are representative of 4 patients. (D) One day after transduction, cells were labeled with 7AAD and annexin V and flow cytometry was performed. Gate frequencies are displayed. Results are representative of 6 patients.