![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.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/3/3/10.1182_bloodadvances.2018027508/2/advances027508f1.png?Expires=1721999840&Signature=ayA4RAwXUUyQYtEXthvgNEp5fdQVbNb6fFk5EYtOr~Ctc6V~0aFjujUkUVMVTIoshkA2PDZX9~0W2alH8UNYpPAkgG2v5He7Js9zTxDWCHUqjnlMDcap1vW6dG1h3unRMiy9XVD7PU9~zuDWp~jrS~wUVp~8N~333MBs1KKX73I~1-BDw4Q7pciEnfyYssxyAQGZh~9s6Tl54ImJAN8bLsI8pwDsMZs9gxpul70tX4y6aCUwozZpw7vPpbqGbA1FX0-PVDdMWo~~5S9ZynjmoQ3ZR1MQGwLUl6XXOQmTrf9pFwAWEgWwmAEvLj0CJEuyc3pQusuDFocaBpyKbLAuDA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
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.