Howard M K, Kershaw T, Gibb B, Storey N, MacLean A R, Zeng B Y, Tel B C, Jenner P, Brown S M, Woolf C J, Anderson P N, Coffin R S, Latchman D S
Department of Molecular Pathology, University College London Medical School, UK.
Gene Ther. 1998 Aug;5(8):1137-47. doi: 10.1038/sj.gt.3300700.
The safe and efficient use of herpes simplex virus (HSV)-based vectors to deliver genes of potentially therapeutic benefit to the central nervous system will require their effective disablement by the inactivation of viral genes required for lytic growth. Here we report that viruses lacking functional genes for ICP27 (which is required for growth in all cell types) and ICP34.5 (which is required for growth in nondividing cell types) can deliver a marker gene to both the rodent and primate CNS with high efficiency whilst producing relatively minimal damage and having no effect on sodium currents in dorsal root ganglion neurons. Such viruses paradoxically deliver genes at much higher efficiency than the less disabled single mutant lacking ICP34.5 alone and also, as expected, produce less damage in vivo. Moreover, unlike the single mutant lacking ICP27 the double mutant viruses cannot revert to wild-type by acquistion of complimenting gene sequences during growth of virus stocks in vitro on dividing cells expressing ICP27 since artificial expression of ICP34.5 in these cells is not required. Such ICP27-; ICP34.5- viruses thus offer a platform for the development of vectors which are sufficiently safe for ultimate use in human gene therapy.
要安全有效地利用基于单纯疱疹病毒（HSV）的载体将具有潜在治疗益处的基因传递至中枢神经系统，就需要通过使裂解生长所需的病毒基因失活来有效灭活这些载体。在此我们报告，缺乏ICP27（所有细胞类型生长所必需）和ICP34.5（非分裂细胞类型生长所必需）功能基因的病毒，能够高效地将标记基因传递至啮齿动物和灵长类动物的中枢神经系统，同时造成相对最小的损伤，且对背根神经节神经元的钠电流没有影响。矛盾的是，此类病毒传递基因的效率比仅缺乏ICP34.5的低失活单突变体要高得多，并且正如预期的那样，在体内造成的损伤也更小。此外，与缺乏ICP27的单突变体不同，双突变病毒在体外表达ICP27的分裂细胞上培养病毒株时，不会通过获取互补基因序列而回复为野生型，因为在这些细胞中不需要人工表达ICP34.5。因此，此类ICP27 - ；ICP34.5 - 病毒为开发最终可安全用于人类基因治疗的载体提供了一个平台。
