title: Analysis of Lujo virus cell entry using pseudotype vesicular stomatitis virus.
abstract: 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 α-dystroglycan [αDG], respectively). However, infection by pseudotype VSV bearing the LUJV envelope protein (LUJpv) occurred independently of hTfR1 and αDG, 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.
  
  
  OBJECTIVE
  LUJV is a newly identified arenavirus associated with a VHF outbreak in southern Africa. Although cell entry for many arenaviruses has been studied, cell entry for LUJV has not been characterized. In this study, we found that LUJpv utilizes neither αDG nor hTfR1 as a receptor and found unique characteristics of LUJV glycoprotein in membrane fusion and cell entry. Proper exclusion of cholesterol or some kinds of lipids may play important roles in LUJpv cell entry.
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Tani
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Iha
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shimojima
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan.
authors.forename: Shuetsu
authors.initials: S
authors.lastname: Fukushi
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Taniguchi
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Yoshikawa
authors.affiliation: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawaoka
authors.affiliation: Department of Biomedical Regulation, School of Health Sciences, Tottori University Faculty of Medicine, Tokyo, Japan.
authors.forename: Naoe
authors.initials: N
authors.lastname: Nakasone
authors.affiliation: Department of Biomedical Regulation, School of Health Sciences, Tottori University Faculty of Medicine, Tokyo, Japan.
authors.forename: Haruaki
authors.initials: H
authors.lastname: Ninomiya
authors.affiliation: Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, University of Tokyo, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Saijo
authors.affiliation: Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo, Japan morikawa@nih.go.jp.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Morikawa
mesh_terms: D000818:Animals; D001117:Arenaviridae Infections / Q000378:metabolism / Q000821:virology*; D002352:Carrier Proteins / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D063927:Lujo virus / Q000254:growth & development* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D000077296:Niemann-Pick C1 Protein; D011956:Receptors, Cell Surface / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism; D054243:Vesicular Stomatitis / Q000378:metabolism / Q000821:virology; D014721:Vesicular stomatitis Indiana virus / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D011956:Receptors, Cell Surface; D013107:Sphingolipids; D014759:Viral Envelope Proteins; D002784:Cholesterol
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