Figure 1.

Systematic improvement of primary human NK cell transduction. Primary human NK cells were isolated and cultured as described in the body of the manuscript. (A) After 7 to 10 days of expansion, NK cells were transduced with serial dilutions of EGFP-expressing lentiviral particles pseudotyped with BaEV-Rless, BaEV-TR, RD114-TR, VSV-G, or GALV-TM. Three to four days after transduction, EGFP expression was analyzed by flow cytometry. Data are represented as mean ± SEM of four biological replicates. (B) Primary human NK cells were transduced with serial dilutions of EGFP-expressing lentiviral particles pseudotyped with BaEV-Rless, BaEV-TR, or GALV-TM, using Retronectin, Vectofusin, or no transduction enhancer, respectively. Three to four days after transduction, EGFP expression was analyzed by flow cytometry. Data are represented as mean ± SEM of five biological replicates. (C) Primary human NK cells were transduced with serial dilutions of lentiviral particles pseudotyped with BaEV-Rless, expressing EGFP under the control of the wild-type or modified SFFV, the wild-type or optimized EF1α, the hPGK, or the MPSV promoter. Three to four days after transduction, the expression of EGFP was analyzed. From the serial dilutions, samples with gene transfer rates of between 5% and 10% were analyzed for their EGFP expression intensity [mean fluorescence intensities (MFIs)]. Data are represented as mean ± SEM of four biological replicates.