Adenovectors are produced by activated E1-CTLs and can transduce LCL targets. There were 2 separate EBV-CTL cell lines transduced with the E1 retrovirus and the Ad5F35-eGFP vector cocultured (at 80:1 LCL/CTL ratio) with autologous (A) or allogeneic (B) LCLs for 48 hours, and then the cells were stained with CD20 or isotype control antibodies. Results are expressed as the percentage of transduced target LCL cells (ie, double positive for GFP and CD20). (C) CTL apoptosis in the presence or absence of LCLs was evaluated using Annexin-V and 7AAD staining of CD3 cells. Cells were stained as recommended by the manufacturer and analyzed by FACS. Results are expressed as percentage in each quadrant. (D) Cytokine production was measured in the supernatant of CTL lines and PBMNCs exposed to 5 × 103 vp of Ad5 or Ad5F35 vector. Cells were transduced as described and supernatants harvested. The cytokines produced from nontransduced CTLs and from transduced PBMNCs were used as negative and positive controls, respectively.
