Development of a lentiviral vector system to study the role of the Andes virus glycoproteins
Development of a lentiviral vector system to study the role of the Andes virus glycoproteins
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DOI：
10.1016/j.virusres.2010.07.001
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发表时间：
2010-10-01
期刊：
影响因子：
5
通讯作者：
Tischler, Nicole D.
中科院分区：
文献类型：
--
作者：
Cifuentes-Munoz, Nicolas;Darlix, Jean-Luc;Tischler, Nicole D.
为了感染靶细胞，包膜病毒使用其病毒体表面蛋白来指导细胞附着并随后通过病毒-细胞膜融合进入。汉坦病毒如何进入细胞在很大程度上还未被探索。为了研究安第斯山脉病毒（ANDV）细胞感染的早期步骤，基于用ANDV-Gn/Gc包膜糖蛋白假型化的猴免疫缺陷病毒（SIV）载体开发了慢病毒载体系统。使用新产生的抗ANDV糖蛋白的单克隆抗体评估Gn和Gc掺入Sly衍生的载体颗粒上。此外，ANDV感染的人的血清能够阻断用ANDV糖蛋白假型化的SIV载体进入细胞，表明它们的抗原构象与天然病毒中的抗原构象相似。用ANDV-Gn/Gc糖蛋白假型化的这种SIV载体的使用将促进关于ANDV细胞进入的研究。沿着这条线，发现从靶细胞中消耗胆固醇强烈地减少了细胞感染，表明脂筏在ANDV细胞进入中的可能作用。Gn/Gc假型SIV载体具有几个优点，特别是高滴度载体生产和通过流式细胞术监测GFP报告基因表达容易定量细胞感染。这种假型SIV载体可用于鉴定Gn/Gc糖蛋白中的功能结构域和筛选潜在的汉坦病毒细胞进入抑制剂。(C)2010 Elsevier B.V.保留所有权利。
To infect target cells, enveloped viruses use their virion surface proteins to direct cell attachment and subsequent entry via virus-cell membrane fusion. How hantaviruses enter cells has been largely unexplored. To study early steps of Andes virus (ANDV) cell infection, a lentiviral vector system was developed based on a Simian immunodeficiency virus (SIV) vector pseudotyped with the ANDV-Gn/Gc envelope glycoproteins. The incorporation of Gn and Gc onto Sly-derived vector particles was assessed using newly generated monoclonal antibodies against ANDV glycoproteins. In addition, sera of ANDV infected humans were able to block cell entry of the SIV vector pseudotyped with ANDV glycoproteins, suggesting that their antigenic conformation is similar to that in the native virus. The use of such SIV vector pseudotyped with ANDV-Gn/Gc glycoproteins should facilitate studies on ANDV cell entry. Along this line, it was found that depletion of cholesterol from target cells strongly diminished cell infection, indicating a possible role of lipid rafts in ANDV cell entry. The Gn/Gc pseudotyped SIV vector has several advantages, notably high titer vector production and easy quantification of cell infection by monitoring GFP reporter gene expression by flow cytometry. Such pseudotyped SIV vectors can be used to identify functional domains in the Gn/Gc glycoproteins and to screen for potential hantavirus cell entry inhibitors. (C) 2010 Elsevier B.V. All rights reserved.