Detection of Ad5 E2B adenoviral DNA in RPMI 8226 MM cells, patient MM2 cells, and normal BM MNCS transduced with Ad.DF3-tk and then treated with GCV. RPMI 8226 MM cells (A) or patient MM2 cells (B; 1 × 103/sample; lanes 1 through 4); RPMI 8226 MM cells (A) or patient MM2 cells (B; 1 × 103/sample) mixed with normal BM MNCs (1 × 104/sample; lanes 5 through 8); or normal BM MNCs (1 × 104/sample; lanes 9 through 12) were either nontransduced (lanes 1, 2, 5, 6, 9, and 10) or transduced with Ad.DF3-tk (MOI = 100; lanes 3, 4, 7, 8, 11, and 12) for 2 hours and then washed out for 10 hours. Cells were next cultured with GCV (50 μmol/L; lanes 2, 4, 6, 8, 10, and 12) or without GCV (lanes 1, 3, 5, 7, 9, and 11) for 36 hours. The E2B gene was amplified by PCR, as previously reported,17 28 from viral DNA extracted following cell lysis. Viral DNA obtained from Ad.DF3-tk particles (1 × 103 to 1 × 105 PFU) served as a positive control (lanes 13 through 15). PCR for β-actin confirmed integrity of DNA.
