Kafri T, van Praag H, Ouyang L, Gage F H, Verma I M
Laboratory of Genetics, The Salk Institute, La Jolla, California 92037, USA.
J Virol. 1999 Jan;73(1):576-84. doi: 10.1128/JVI.73.1.576-584.1999.
Lentivirus vectors can transduce dividing and nondividing cells. Using three-plasmid transient transfections, high-titer (>10(9) IU/ml) recombinant lentivirus vectors pseudotyped with vesicular stomatitis virus G (VSV-G) protein can be generated (T. Kafri et al., Nat. Genet. 17:314-317, 1997; H. Miyoshi et al., Proc. Natl. Acad. Sci. USA 94:10319-10323, 1997; L. Naldini et al., Science 272:263-267, 1996). The recombinant lentiviruses can efficiently infect brain, liver, muscle, and retinal tissue in vivo. Furthermore, the transduced tissues demonstrated long-term expression of reporter genes in immunocompetent rodents. We now report the generation of a tetracycline-inducible VSV-G pseudotyped lentivirus packaging cell line which can generate virus particles at titers greater than 10(6) IU/ml for at least 3 to 4 days. The vector produced by the inducible cell line can be concentrated to titers of 10(9) IU/ml and can efficiently transduce nondividing cells in vitro and in vivo. The availability of a lentivirus packaging cell line will significantly facilitate the production of high-titer lentivirus vectors for gene therapy and study of human immunodeficiency virus biology.
慢病毒载体能够转导分裂细胞和非分裂细胞。通过三质粒瞬时转染，可以产生用泡状口炎病毒G（VSV-G）蛋白假型化的高滴度（>10⁹ IU/ml）重组慢病毒载体（T. 卡夫里等人，《自然遗传学》17:314 - 317，1997；H. 宫城等人，《美国国家科学院院刊》94:10319 - 10323，1997；L. 纳尔迪尼等人，《科学》272:263 - 267，1996）。重组慢病毒能够在体内高效感染脑、肝、肌肉和视网膜组织。此外，在有免疫活性的啮齿动物中，转导的组织显示出报告基因的长期表达。我们现在报告一种四环素诱导型VSV-G假型化慢病毒包装细胞系的产生，该细胞系能够产生滴度大于10⁶ IU/ml的病毒颗粒，至少持续3至4天。由诱导型细胞系产生的载体可以浓缩至10⁹ IU/ml的滴度，并且能够在体外和体内高效转导非分裂细胞。慢病毒包装细胞系的可用性将显著促进用于基因治疗和人类免疫缺陷病毒生物学研究的高滴度慢病毒载体的生产。
