H/F-LVs allow genuine transduction of quiescent B cells. (A) Quiescent adult B cells were transduced with H/F-LVs (MOI = 10) or VSVG-LVs (MOI = 30) in the absence or presence of AZT. The dot plots show the percentage of GFP in the CD19⁺ population at day 2 after transduction. (B) Cells were transduced with an H/F-LV encoding a nonfunctional integrase (INT-D116A) or a wild-type integrase (INT-wt) at an MOI of 10. GFP expression and MFI are shown for day 2 after transduction. (C) Quiescent B cells were transduced with H/F-LVs (MOI = 10) or VSVG-LVs (MOI = 30). At day 2 after transduction, part of the B cells was continued on a monolayer of MS5 cells. GFP-expression in the B-cell (CD19⁺) population is indicated for day 2, day 6, and day 12 after transduction as analyzed by FACS (means ± SD; n = 3). A nested Alu-PCR on the DNA of transduced and nontransduced sorted CD19⁺ cells at day 12 of culture on MS5 cells was performed using specific primers (see “Alu-PCR”). The PCR-specific band corresponding to integrated vector sequence of 121 base pair is detected only in H/F-transduced B cells (graph inset).