Simultaneous induction of trivirus
cells. (A) A diagram depicting the standard protocol for the generation of triVSTs. (B) Comparison of two protocols for the simultaneous induction of triVSTs. In Ad/PBMC + LCL protocol, PBMCs were cultured in non-tissue culture treated plates overnight then transduced with Ad5f35-pp65 and cocultured with autologous LCLs. In the Ad/LCL protocol, LCLs were transduced with the Ad5f35-pp65 vector and used for autologous PBMC stimulation. (C) Day 9 after the first stimulation, the AdV (hexon and penton), CMV (pp65) and EBV (Auto LCL) specificities were determined by ELIspot assay using pepmixes for hexon, penton and pp65 and autologous LCLs for EBV. The MOI of Ad5f35-pp65 transduction and the ratio of PBMC to LCL are indicated. (D) TriVSTs were generated using Ad/LCLs and Ad/PBMC + LCLs (100:1) and counted on days 3, 10, and 14. Data represent the mean +/− sd of 2 donors.