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. 2019 Feb 12;3(3):461–475. doi: 10.1182/bloodadvances.2018027508

Figure 1.

Figure 1.

BaEV-LVs allow high-level transduction of naive as well as memory adult human T cells. (A) Freshly isolated peripheral blood T cells were preactivated through the TCR (anti-CD3 + anti-CD28 antibodies + IL-2) for 24 hours and transduced in the absence and presence of RetroNectin with the different LV pseudotypes at indicated MOIs. Six days after transduction, the percentage of GFP+CD3+ T cells was analyzed by FACS (mean ± standard deviation [SD]; n = 4). (B-C) Freshly isolated CD3+ T cells were prestimulated with IL-7 for 3 days and transduced in the presence of RetroNectin with the different LV pseudotypes at MOI 10 or 100 as indicated. Six days after transduction, the percentage of GFP+CD3+ T cells was analyzed by FACS for total T cells (anti-CD3) and the naive (CD45RA+) or memory (anti-CD45RO+) T cells subsets in panel B. (C) More detailed phenotyping of these cells shows the percentage of GFP+ naive (CD45RA+CD62L+CCR7+) CD4 and CD8 T cells. (B-C; mean ± SD; n = 3; VSV-G-LV MOI 10 vs BaEV-LV). (D) ASCT-1 and ASCT-2 expression levels in unstimulated (unstim.) and TCR- or IL-7–activated total CD3+ T cells (as in panels A and B, respectively). Relative fold increase in ASCT-1 or ASCT-2 mRNA levels in stimulated (stim.) CD3+ T cells compared with unstimulated T cells, for which mRNA levels were normalized to 1 (mean ± SD; n = 3). *P < .05; **P < .01. ns, nonsignificant.