The HΔ24/FΔ30-displaying LVs efficiently transduce quiescent adult T cells. (A) Highly purified resting adult T cells were isolated by negative antibody-mediated selection to avoid cell activation and then transduced with HΔ24/FΔ30- or VSV-G–pseudotyped HIV-GFP vectors at an MOI of 10 or 30 in the absence of exogenous stimuli. Three days after transduction, surface staining for naive and memory subsets was performed by anti-CD45RA/anti-CD45RO double staining. In parallel, surface staining for the CD3⁺, CD4⁺, and CD8⁺ T-cell subsets was performed and the percentage of GFP expression for each of these T-cell subsets was analyzed by FACS (means ± SD; n = 5). Inset shows CD45RA versus SLAM expression on freshly isolated total CD3⁺ T cells (B) Resting T cells were transduced with HΔ24/FΔ30- or VSV-G–pseudotyped LVs at an MOI of 10. After 3 days, part of the transduced cell cultures was continued in the presence of rIL-7 for 3 more days to verify stable GFP expression. For both culture conditions, surface staining for naive and memory subsets was performed by anti-CD45RA/anti-CD45RO double staining. Dot blots represent the percentage of GFP⁺ cells in the CD45RA⁺ naive T cells (top row) and the percentage GFP⁺ cells in the CD45RO⁺ memory T cells (bottom row). Data are representative of 6 experiments. (C) Resting T cells were transduced with HΔ24/FΔ30- or VSV-G–pseudotyped LVs at an MOI of 30. Surface staining for naive T cells was performed by anti-CD45RA staining. Dot blot represents the GFP⁺ CD45RA⁺ naive T cells (top right quadrant R2) and the GFP⁺ CD45RA⁻ memory T cells (bottom right quadrant R4). For each gate, R1, R2, R3, and R4, the expression of CD62L is shown. Data are representative of 3 experiments. (D) Surface expression of SLAM or CD46 receptors on resting adult CD3⁺ cells after transduction with HΔ24/FΔ30- or VSV-G–pseudotyped lentivectors at an MOI of 10. (E) Resting adult T cells were incubated with or without anti-SLAM or anti–CD46 receptor–blocking antibodies for 3 hours and subsequently transduced with HΔ24/FΔ30-LVs or VSV-G-LVs at an MOI of 10. The percentage of GFP⁺ cells was analyzed by FACS 3 days after transduction.