FIG. 2.

Transduction of growth-arrested HeLa cells and of primary human blood cells with HIV-1 and F-MLV vectors. VSVg-pseudotyped vectors were purified on a double-sucrose cushion after production from 293T cells, and normalized amounts of HeLa infectious particles were used on target cells at different MOIs (as indicated). The percentage of GFP-positive cells was scored 3 to 4 days later by flow cytometry. The different primary cells shown here were derived from the same donor, and average results from three to seven different donors are presented. HeLa cells were arrested in either G₂/M by gamma irradiation (γ-irr; 6,000 rads) or in G₁/S by aphidicolin (aphi) treatment (10 μg/ml), and PBLs were treated with PHA plus IL-2 (to induce cell proliferation) or IL-7 (to induce G₀ to G_1b transition) 24 h prior to transduction. Human blood monocytes were differentiated in macrophages and DCs in presence of GM-CSF and GM-CSF plus IL-4, respectively, and transduced between days 4 and 6. When indicated, AZT and ddI were added to cells prior to viral transduction.