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用泡状口炎病毒糖蛋白假型化的病毒进行逆转录转导。

Retro-transduction by virus pseudotyped with glycoprotein of vesicular stomatitis virus.

作者信息

Ohishi Masahisa, Shioda Tatsuo, Sakuragi Jun-ichi

机构信息

Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita City, Osaka 565-0871, Japan.

出版信息

Virology. 2007 May 25;362(1):131-8. doi: 10.1016/j.virol.2006.12.030. Epub 2007 Jan 26.

DOI:10.1016/j.virol.2006.12.030
PMID:17258261
Abstract

A virus pseudotyped with glycoprotein of vesicular stomatitis virus (VSV-G) can enter various cell types at a relatively high titer. We observed that the amount of viral antigen from VSV-G pseudotyped human immunodeficiency virus type 1 (HIV-1) producing cells was much higher than that from their non-pseudotyped counterparts. This enhanced viral antigen production was not observed when we used HIV-1 pol mutant, viral enzyme inhibitors, HIV Env protein, or VSV-G fusion defective mutants. The transfection experiment using GFP-expressing virus showed time-dependent expansion of GFP-positive cells and viral DNA integration. These results suggested that the increase in viral antigen yield was caused by the release of a progeny virus following retro-transduction by the pseudotyped virus of the cells within the transfected cell culture. The infectivity as well as the amount of VSV-G on virus particles per unit of viral antigen was significantly different before and after the onset of the yield enhancement. This suggests that results of infection assays of the virus pseudotyped with VSV-G may be affected by the occurrence of such enhancement. This means that, while pseudotyping with VSV-G is a simple and effective method, this procedure should be carefully considered when the virus is produced for infectivity assays.

摘要

一种用水泡性口炎病毒(VSV-G)糖蛋白假型化的病毒能够以相对较高的滴度进入多种细胞类型。我们观察到,来自产生VSV-G假型化1型人类免疫缺陷病毒(HIV-1)的细胞的病毒抗原量远高于其未假型化的对应细胞。当我们使用HIV-1 pol突变体、病毒酶抑制剂、HIV Env蛋白或VSV-G融合缺陷突变体时,未观察到这种病毒抗原产生的增强。使用表达绿色荧光蛋白(GFP)的病毒进行的转染实验显示,GFP阳性细胞呈时间依赖性扩增且病毒DNA发生整合。这些结果表明,病毒抗原产量的增加是由于在转染细胞培养物中,假型化病毒对细胞进行逆转录转导后子代病毒的释放所致。在产量增强开始前后,每单位病毒抗原的病毒颗粒上VSV-G的感染性以及量存在显著差异。这表明,用VSV-G假型化的病毒的感染测定结果可能会受到这种增强现象的影响。这意味着,虽然用VSV-G进行假型化是一种简单有效的方法,但在为感染性测定生产病毒时,应谨慎考虑这一过程。

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