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首页> 外文期刊>Biochemical Engineering Journal >Cholesterol supplementation during production increases the infectivity of retroviral and lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein(VSV-G)
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Cholesterol supplementation during production increases the infectivity of retroviral and lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein(VSV-G)

机译：生产过程中补充胆固醇会增加水泡性口腔炎病毒糖蛋白（VSV-G）假型的逆转录病毒和慢病毒载体的感染性

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摘要

Cholesterol,a major component of plasma membrane lipid rafts,is important for assembly and budding of enveloped viruses,including influenza and HIV-1.Cholesterol depletion impairs virus assembly and infectivity.This study examined the effects of exogenous cholesterol addition(delivered as a complex with methyl-beta-cyclodextrin(MbCD))on the production of Molony murine leukemia virus(MoMuLV)retroviral vector and HIV-1-based lentiviral vector pseudotyped with the vesicular stomatitis virus glycoprotein(VSV-G).Cholesterol supplementation before and during vector production enhanced the infectivity of retroviral and lentiviral vectors up to 4-fold and 6-fold,respectively.In contrast,the amount of retroviral vector produced was unchanged,and that of lentiviral vector was increased less than 2-fold.Both free cholesterol and cholesterol ester content in 293-gag-pol producer cells increased with cholesterol addition.In contrast,the phospholipids headgroup composition was essentially unchanged by cholesterol supplementation in 293-gag-pol packaging cells.Based on these results,it is proposed that cholesterol supplementation increases the infectivity of VSV-G-pseudotyped retroviral and lentiviral vectors,possibly by altering the composition of the producer cell membrane where the viral vectors are assembled and bud,and/or by changing the lipid composition of the viral vectors.
机译：胆固醇是质膜脂质筏的主要成分，对于包膜病毒（包括流感和HIV-1）的组装和萌发非常重要。胆固醇的消耗会削弱病毒的组装和感染性。本研究研究了外源胆固醇添加的影响（以复合物形式提供）甲基-β-环糊精（MbCD）合成Molony鼠白血病病毒（MoMuLV）逆转录病毒载体和以水疱性口腔炎病毒糖蛋白（VSV-G）假型的基于HIV-1的慢病毒载体的生产。生产过程中，逆转录病毒和慢病毒载体的感染力分别提高了4倍和6倍。相比之下，逆转录病毒载体的产生量没有变化，而慢病毒载体的增加量不到2倍。游离胆固醇和293-gag-pol生产者细胞中的胆固醇酯含量随胆固醇的添加而增加。相比之下，磷脂的头基组成基本没有变化根据这些结果，有人建议通过补充胆固醇来增加VSV-G-假型逆转录病毒和慢病毒载体的感染力，可能是通过改变病毒的生产细胞膜的组成来实现的。组装和萌芽载体，和/或通过改变病毒载体的脂质组成。

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