A system for functional analysis of Ebola virus glycoprotein
Ayato Takada,Clinton S. Robison,Hideo Goto,Anthony Sanchez,K. Gopal Murti,Michael A. Whitt,Yoshihiro Kawaoka +6 more
TLDR
It is suggested that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry.Abstract:
Ebola virus causes hemorrhagic fever in humans and nonhuman primates, resulting in mortality rates of up to 90%. Studies of this virus have been hampered by its extraordinary pathogenicity, which requires biosafety level 4 containment. To circumvent this problem, we developed a novel complementation system for functional analysis of Ebola virus glycoproteins. It relies on a recombinant vesicular stomatitis virus (VSV) that contains the green fluorescent protein gene instead of the receptor-binding G protein gene (VSVΔG*). Herein we show that Ebola Reston virus glycoprotein (ResGP) is efficiently incorporated into VSV particles. This recombinant VSV with integrated ResGP (VSVΔG*-ResGP) infected primate cells more efficiently than any of the other mammalian or avian cells examined, in a manner consistent with the host range tropism of Ebola virus, whereas VSVΔG* complemented with VSV G protein (VSVΔG*-G) efficiently infected the majority of the cells tested. We also tested the utility of this system for investigating the cellular receptors for Ebola virus. Chemical modification of cells to alter their surface proteins markedly reduced their susceptibility to VSVΔG*-ResGP but not to VSVΔG*-G. These findings suggest that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry. Thus, our VSV system should be useful for investigating the functions of glycoproteins from highly pathogenic viruses or those incapable of being cultured in vitro.read more
Citations
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Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses.
TL;DR: An approach to rapidly screen lineage B betacoronaviruses, such as SARS-CoV and the recently emerged SARS -CoV-2, for receptor usage and their ability to infect cell types from different species is developed.
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Ebola virus entry requires the cholesterol transporter Niemann–Pick C1
Jan E. Carette,Jan E. Carette,Matthijs Raaben,Anthony C. Wong,Andrew S. Herbert,Gregor Obernosterer,Gregor Obernosterer,Nirupama Mulherkar,Ana I. Kuehne,Philip J. Kranzusch,April M. Griffin,Gordon Ruthel,Paola Dal Cin,John M. Dye,Sean P. J. Whelan,Kartik Chandran,Thijn R. Brummelkamp,Thijn R. Brummelkamp +17 more
TL;DR: It is shown that membrane fusion mediated by filovirus glycoproteins and viral escape from the vesicular compartment require the NPC1 protein, independent of its known function in cholesterol transport, which indicates potential antiviral strategies to combat these deadly agents.
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Antibody cocktail to SARS-CoV-2 spike protein prevents rapid mutational escape seen with individual antibodies.
Alina Baum,Benjamin O. Fulton,Elzbieta Wloga,Richard Copin,Kristen E. Pascal,Vincenzo Russo,Stephanie Giordano,Kathryn Lanza,Nicole Negron,Min Ni,Yi Wei,Gurinder S. Atwal,Andrew J. Murphy,Neil Stahl,George D. Yancopoulos,Christos A. Kyratsous +15 more
TL;DR: This work investigated the development of resistance against four antibodies to the spike protein that potently neutralize SARS-CoV-2, individually as well as when combined into cocktails.
Journal ArticleDOI
Studies in humanized mice and convalescent humans yield a SARS-CoV-2 antibody cocktail.
Johanna Hansen,Alina Baum,Kristen E. Pascal,Vincenzo Russo,Stephanie Giordano,Elzbieta Wloga,Benjamin O. Fulton,Ying Yan,Katrina Koon,Krunal Patel,Kyung Min Chung,Aynur Hermann,Erica Ullman,Jonathan W. Cruz,Ashique Rafique,Tammy T. Huang,Jeanette L. Fairhurst,Christen Libertiny,Marine Malbec,Wen-Yi Lee,Richard Welsh,Glen Farr,Seth Pennington,Dipali Deshpande,Jemmie Cheng,Anke Watty,Pascal Bouffard,Robert Babb,Natasha Levenkova,Calvin R. Chen,Bojie Zhang,Annabel Romero Hernandez,Kei Saotome,Yi Zhou,Matthew C. Franklin,Sumathi Sivapalasingam,David C. Lye,Stuart Weston,James Logue,Robert Haupt,Matthew B. Frieman,Gang Chen,William C. Olson,Andrew J. Murphy,Neil Stahl,George D. Yancopoulos,Christos A. Kyratsous +46 more
TL;DR: In this article, the authors describe parallel efforts using both humanized mice and convalescent patients to generate antibodies against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein, which yielded a large collection of fully human antibodies that were characterized for binding, neutralization and three-dimensional structure.
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SLAM (CDw150) is a cellular receptor for measles virus.
TL;DR: It is shown that human SLAM (signalling lymphocyte-activation molecule), a recently discovered membrane glycoprotein expressed on some T and B cells, is a cellular receptor for measles virus, including the Edmonston strain.
References
More filters
Journal ArticleDOI
Green fluorescent protein as a marker for gene expression
TL;DR: A complementary DNA for the Aequorea victoria green fluorescent protein produces a fluorescent product when expressed in prokaryotic or eukaryotic cells, which can be used to monitor gene expression and protein localization in living organisms.
Journal ArticleDOI
Efficient selection for high-expression transfectants with a novel eukaryotic vector
TL;DR: The results showed that high concentrations of G418 efficiently yielded L cell and CHO cell transfectants stably producing IL-2 at levels comparable with those previously attained using gene amplification.
Journal ArticleDOI
Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase
TL;DR: The vaccinia/T7 hybrid virus forms the basis of a simple, rapid, widely applicable, and efficient mammalian expression system.
PatentDOI
Herpes simplex virus 1 entry into cells mediated by a novel member of the tnf/ngf receptor family
TL;DR: HVEM, the first identified mediator of HSV entry, is a new member of the TNF/NGF receptor family and showed to mediate the entry of several wild-type HSV strains of both serotypes.
Journal ArticleDOI
Recombinant vesicular stomatitis viruses from DNA
TL;DR: The ability to generate VSV from DNA opens numerous possibilities for the genetic analysis of VSV replication and may be possible to genetically engineer recombinant VSVs displaying foreign antigens, which could be useful as vaccines conferring protection against other viruses.