CD3-LVs transduce T cells independently of activation or cytokine treatment. (A-B) Primary human PBMCs isolated from different blood donations were activated with αCD3 and αCD28 antibodies for 3 days in the presence of IL-2 before they were incubated with CD3-LVs or VSV-LV. Green fluorescent protein (GFP) expression was determined 6 days later by flow cytometry. (A) Representative dot plots show GFP expression in CD4⁺ and CD4⁻ populations gated for viable cells. (B) Scatter bar diagrams summarize the percentages of GFP⁺ cells as mean ± standard deviation (SD) of 3 independent experiments with 2-3 donors and 2-3 technical replicates. (C-D) Freshly isolated PBMCs were cultured overnight in presence of IL-2 or IL-7/IL-15, or in absence of cytokines, before they were transduced with CD3-LVs or VSV-LV. GFP expression was determined 6 days later by flow cytometry. (C) Scatter bar diagrams show the percentage of GFP⁺ cells in all viable single cells. (D) Representative dot plots show GFP expression in the CD8⁺ and CD8⁻ populations gated for viable cells. Data are mean ± SD from 1 experiment with n = 3 technical replicates; biological replicates from additional experiments with IL-2- and IL-7/IL-15-stimulated cells are shown in supplemental Figure 6C-F. **P < .01, ***P < .001, ****P < .0001 by 2-way analysis of variance (ANOVA) with Dunnett's correction (comparison with VSV-LV for each condition), or unpaired Student t test.