Nguyen T H, Pagès J C, Farge D, Briand P, Weber A
Institut Cochin de Génétique Moléculaire, Inserm U380, Paris, France.
Hum Gene Ther. 1998 Nov 20;9(17):2469-79. doi: 10.1089/hum.1998.9.17-2469.
The development of retroviral vectors with cell-specific targeting capabilities will be an important step toward successful in vitro gene therapy. This article describes the generation of a retroviral vector with enhanced binding abilities for cells bearing the c-Met receptor: the Madin-Darby canine kidney (MDCK) cell line and primary hepatocytes. The human hepatocyte growth factor (HGF) was displayed on murine amphotropic retroviral vectors by fusion to the viral transmembrane envelope glycoprotein (TM). The resulting chimeric envelope HGF-TM was expressed in an amphotropic packaging cell line producing viral particles that display both HGF-TM and the wild-type envelope. These modified viral particles had a titer equivalent to that of unmodified particles. Modified particles infected MDCK cells more efficiently than did unmodified amphotropic retrovirus. Adding anti-HGF antibodies to the viral vector particle supernatant prior to infection confirmed that the increased infection was mediated by the HGF moiety. The chimeric viruses also infected primary mouse and nonhuman primate fetal hepatocytes more effectively. Furthermore, these cells could be induced to proliferate by the modified HGF-TM viruses. Since exogenous HGF is primarily taken up by the liver, these results may have implications for retroviral vector design for liver-directed human gene therapy.
开发具有细胞特异性靶向能力的逆转录病毒载体将是迈向成功体外基因治疗的重要一步。本文描述了一种对携带c-Met受体的细胞（如犬肾细胞系（MDCK）和原代肝细胞）具有增强结合能力的逆转录病毒载体的构建。通过与病毒跨膜包膜糖蛋白（TM）融合，将人肝细胞生长因子（HGF）展示在鼠嗜异性逆转录病毒载体上。所得嵌合包膜HGF-TM在嗜异性包装细胞系中表达，产生同时展示HGF-TM和野生型包膜的病毒颗粒。这些修饰后的病毒颗粒滴度与未修饰颗粒相当。修饰后的颗粒比未修饰的嗜异性逆转录病毒更有效地感染MDCK细胞。在感染前向病毒载体颗粒上清液中加入抗HGF抗体，证实感染增加是由HGF部分介导的。嵌合病毒也能更有效地感染原代小鼠和非人类灵长类动物胎儿肝细胞。此外，这些细胞可被修饰后的HGF-TM病毒诱导增殖。由于外源性HGF主要被肝脏摄取，这些结果可能对用于肝脏定向人类基因治疗的逆转录病毒载体设计具有启示意义。
