Development of herpes simplex virus replication-defective multigene vectors for combination gene therapy applications
Development of herpes simplex virus replication-defective multigene vectors for combination gene therapy applications
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DOI：
10.1038/sj.gt.3300755
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发表时间：
1998-11-01
期刊：
影响因子：
5.1
通讯作者：
Glorioso, JC
中科院分区：
文献类型：
--
作者：
Krisky, DM;Marconi, PC;Glorioso, JC
一些基因治疗应用将需要同时表达多种基因产物以实现治疗效果。在这项研究中，我们描述了复制缺陷型单纯疱疹病毒1型（HSV-1）载体（HX 86 Z或HX 86 G）的产生和表征，携带不同的和独立调控的表达盒的五个转基因（hIL-2，hGM-CSF，hB7.1，HSV-tk和lacZ或hIFN γ）。将代表12 kb DNA序列的转基因重组到单个突变病毒载体的单独基因座中，缺失代表九个病毒基因ICP 4、ICP 22、ICP 27、ICP 47、U（L）24、U（L）41、U（L）44、U（s）10和Us（11）的部分的11.6kb载体序列。即刻早期基因ICP 4、ICP 22和ICP 27的缺失显著降低了载体细胞毒性，阻止了早期和晚期病毒基因表达，并保留了完整的MHC I类抗原表达。在原代人黑素瘤细胞中获得多个转基因的同时表达长达7天，在感染后2-3天达到峰值表达。选择转基因是因为它们在肿瘤破坏和疫苗基因治疗应用中具有协同作用的潜力，但是所采用的方法和载体可以应用于其他多基因治疗策略。这项研究表明，工程化的大转基因容量的DNA病毒，如HSV-1的表达多种转基因的潜力。
Some gene therapy applications will require simultaneous expression of multiple gene products to achieve a therapeutic effect. In this study we describe the generation and characterization of replication incompetent herpes simplex virus type 1 (HSV-1) vectors (HX86Z or HX86G) carrying distinct and independently regulated expression cassettes for five transgenes (hIL-2, hGM-CSF, hB7.1, HSV-tk and lacZ or hIFN gamma). The transgenes, representing 12 kb of DNA sequence, were recombined into separate loci of a single mutant virus vector deleted for 11.6 kb of vector sequences representing portions of nine viral genes, ICP4, ICP22, ICP27, ICP47, U(L)24, U(L)41, U(L)44, U(s)10 and Us(11). Deletion of the immediate-early genes ICP4, ICP22 and ICP27 substantially reduced vector cytotoxicity, prevented early and late viral gene expression and left intact MHC class I antigen expression. Simultaneous expression of multiple transgenes was obtained for up to 7 days in primary human melanoma cells with peak expression at 2-3 days after infection. The transgenes were chosen for their potential to function synergistically in tumor destruction and vaccine gene therapy applications, but the method and vector employed could be applied to other multigene therapy strategies. This study demonstrates the potential for engineering large transgene capacity DNA viruses such as HSV-1 for expression of multiple transgenes.