Efficiency of Ad vector transduction assessed byβ-gal reporter gene expression in OCI-My5 and RPMI 8226 MM cells, as well as RPMI 8226 MM cells within normal BM MNCs. OCI-My5 and RPMI 8226 MM cells (5 × 105/mL) were transduced with Ad.DF3-βgal at MOI = 1 for 2 hours, incubated with FDG, and analyzed by fluorescence flow cytometry at 24 hours (A). These tumor cells were also transduced with Ad.DF3-βgal (▪) or Ad.CMV-βgal (□) at MOI = 10 for 2 hours and analyzed by chemiluminescence assay at 24 hours (B). Mock-irradiated (□) and γ-irradiated (▪; 2.0 Gy) normal BM MNCs (3 × 104/well) were cultured in media alone or transduced with either Ad.CMV-βgal or Ad.DF3-βgal (both MOI = 100) for 2 hours and then analyzed by X-Gal staining at 24 hours (n = 3). X-Gal staining of mock-irradiated and γ-irradiated BM MNCs was compared with untransduced normal BM MNCs (negative control) and RPMI 8226 MM cells (positive control; C). RPMI 8226 MM cells (0.0%, 0.0001%, 0.001%, and 0.01%) mixed with normal BM MNCs were transduced with Ad.DF3-βgal at MOI = 1 for 2 hours and similarly analyzed using a chemiluminescence assay at 24 hours (n = 3; D).
