Optimization of the induction of trivirus specificity with Ad5f35
pp65 transduced LCL. (A) Trivirus specificity was determined by γ-IFN ELIspot assay 9 days after the 1st stimulation. The MOIs of Ad5f35-pp65 transduction of LCLs were 103, 104, and 105 vp per cell respectively. (B) LCLs were transduced with an MOI of 104 and cultured for the indicated times before being irradiated and used for PBMC stimulation. ELIspot assays were performed 9 days after the 1st stimulation. Non-transduced (NT) LCLs were used as controls. Data represent the mean +/− sd of 2 donors. (C) Tri-virus specific CTLs were initiated on Day 0 and re-stimulated on Days 9 and 18. Tri-virus specificities were assessed by γ-IFN ELIspot assay with LCLs or overlapping libraries of pepmixes spanning adenovirus hexon and penton and CMV pp65 proteins.