Analysis of Lujo Virus Cell Entry using Pseudotype Vesicular Stomatitis Virus
Analysis of Lujo Virus Cell Entry using Pseudotype Vesicular Stomatitis Virus
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DOI：
10.1128/jvi.00512-14
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发表时间：
2014-07-01
影响因子：
5.4
通讯作者：
Morikawa, Shigeru
中科院分区：
文献类型：
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作者：
Tani, Hideki;Iha, Koichiro;Morikawa, Shigeru
已知几种沙粒病毒在撒哈拉以南非洲和南美洲引起病毒性出血热（VHF），其中VHF是一个主要的公共卫生和医学问题。这些沙粒病毒的生物安全4级分类限制了它们的使用，并阻碍了生物学研究，包括治疗药物和/或疫苗的开发。由于与处理活病毒相关的困难，已经开发了假型病毒作为替代病毒系统，所述假型病毒瞬时携带基于水泡性口炎病毒（VSV）或逆转录病毒的沙粒病毒包膜蛋白。在这里，我们报告了一个假型VSV轴承每个包膜蛋白的各种沙粒病毒（AREpv），包括新发现的卢霍病毒（LUJV）和查帕雷病毒的发展。在293 T细胞中产生的假型沙粒病毒在各种哺乳动物细胞系中表现出高感染性。新世界和旧世界AREpv的感染依赖于它们的受体（分别为人转铁蛋白受体1 [hTfR 1]和α-dystro-聚糖[alpha DG]）。然而，由假型VSV轴承的LUJV包膜蛋白（LUJpv）的感染发生独立的hTfR 1和α DG，表明LUJpv利用一个未知的受体。通过使用亲溶酶体剂证实了AREpv的pH依赖性内吞作用。表达这些包膜蛋白的细胞的融合，除了那些表达LUJV包膜蛋白，诱导在低pH值的瞬时处理。LUJpv的感染性被胆固醇转运抑制剂U18666 A抑制。此外，LUJpv的感染性在尼曼-匹克C1（NPC 1）缺陷细胞系中显著降低，表明NPC 1活性对于有效的LUJpv感染是必要的。
Several arenaviruses are known to cause viral hemorrhagic fever (VHF) in sub-Saharan Africa and South America, where VHF is a major public health and medical concern. The biosafety level 4 categorization of these arenaviruses restricts their use and has impeded biological studies, including therapeutic drug and/or vaccine development. Due to difficulties associated with handling live viruses, pseudotype viruses, which transiently bear arenavirus envelope proteins based on vesicular stomatitis virus (VSV) or retrovirus, have been developed as surrogate virus systems. Here, we report the development of a pseudotype VSV bearing each envelope protein of various species of arenaviruses (AREpv), including the newly identified Lujo virus (LUJV) and Chapare virus. Pseudotype arenaviruses generated in 293T cells exhibited high infectivity in various mammalian cell lines. The infections by New World and Old World AREpv were dependent on their receptors (human transferrin receptor 1 [hTfR1] and alpha-dystro-glycan [alpha DG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and alpha DG, indicating that LUJpv utilizes an unidentified receptor. The pH-dependent endocytosis of AREpv was confirmed by the use of lysosomotropic agents. The fusion of cells expressing these envelope proteins, except for those expressing the LUJV envelope protein, was induced by transient treatment at low pH values. LUJpv infectivity was inhibited by U18666A, a cholesterol transport inhibitor. Furthermore, the infectivity of LUJpv was significantly decreased in the Niemann-Pick C1 (NPC1)-deficient cell line, suggesting the necessity for NPC1 activity for efficient LUJpv infection.