Retroviral Transduction

HepG2 cells were seeded in a 24-well tissue culture plate to yield 70%–80% confluency. 2 mL of 0.45 μm-filtered respective retroviral supernatants were applied to each well and incubated for 48 hr at 37°C in 5% CO₂. HCV-GFP⁺ positive cells were sorted for high and uniform expression of GFP using a FACSAria III^u instrument (BD Biosciences, San Jose, CA). Activated T cells were transduced to express the HCV1406 TCR by spinoculation as previously described.6, 15, 58, 59, 60, 61 T cells were then sorted for TCR-transduced cells by positive selection using anti-CD34 mAb-coated immunomagnetic beads (Miltenyi Biotec) and used in functional assays.