Adoptive Transfer of Activated T Cells

HEK293T cells were transfected and treated as described above. On day 2, splenocytes were prepared from STG mice and cultured in 10-cm plates at a density of 5–10 × 10⁷ cells in 15 ml T cell media with soluble LCMV peptide GP61-80 (1 μg/ml; AnaSpec). The following day, activated STG⁺ T cells were isolated using an EasySep Mouse CD4⁺ T cell isolation kit (Stemcell Technologies) and plated in 24-well plates at a density of 1–2 × 10⁶ cells per well. Cells were transduced with viral supernatants as described above. Following transduction, cells were washed and resuspended in PBS for adoptive transfer via retro-orbital injection into recipient mice.