Gallardo H F, Tan C, Ory D, Sadelain M
Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Blood. 1997 Aug 1;90(3):952-7.
It is essential for the study of T-cell function and the improvement of adoptive cell therapies to efficiently generate large populations of human primary T cells that reliably express foreign genes. This goal is achieved by using recombinant retroviruses pseudotyped with either the gibbon ape leukemia virus (GaLV) envelope or the vesicular stomatitis virus G (VSV-G) glycoprotein. We show here that both retroviral particles mediate stable gene transfer in CD4+ and in CD8+ peripheral blood lymphocytes cultured under optimized conditions. However, VSV-G-pseudotyped virions may cause transduction artifacts that must be carefully excluded. The VSV-G virions require 10- to 100-fold higher concentrations of infectious particles to achieve levels of gene transfer comparable to GaLV-virions. Nonetheless, the physical stability of VSV-G-coated particles enables the concentration of viral stocks to 10(9) infectious particles per milliliter or more.
高效生成大量可靠表达外源基因的人原代T细胞，对于T细胞功能研究和过继性细胞疗法的改进至关重要。通过使用携带长臂猿白血病病毒（GaLV）包膜或水泡性口炎病毒G（VSV-G）糖蛋白的重组逆转录病毒假型可实现这一目标。我们在此表明，在优化条件下培养的CD4+和CD8+外周血淋巴细胞中，两种逆转录病毒颗粒均介导稳定的基因转移。然而，VSV-G假型病毒粒子可能会导致必须仔细排除的转导假象。VSV-G病毒粒子需要浓度高10至100倍的感染性颗粒才能达到与GaLV病毒粒子相当的基因转移水平。尽管如此，VSV-G包膜颗粒的物理稳定性使病毒储备液的浓度能够达到每毫升10^9个感染性颗粒或更高。
