Abstract
Members of the family Filoviridae are filamentous, enveloped, and nonsegmented negative-stranded RNA viruses that can cause severe hemorrhagic disease in humans and nonhuman primates with high mortality rates. Current efforts to analyze the structure and biology of these viruses as well as the development of antivirals have been hindered by the necessity of biosafety level 4 containment (BSL4). Here, we outline how to produce and work with Ebola virus glycoprotein bearing vesicular stomatitis virus (VSV) pseudovirions. These pseudovirions can be safely used to evaluate early steps of the filovirus life cycle without need for BSL4 containment. Virus gene expression in the transduced cells is easy to assess since the pseudovirions encode a reporter gene in place of the VSV G glycoprotein gene. Adoption of VSV for use as a pseudovirion system for filovirus GP has significantly expanded access for researchers to study specific aspects of the viral life cycle outside of BSL4 containment and has allowed substantial growth of filovirus research.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Feldmann H, Klenk HD, Sanchez A (1993) Molecular biology and evolution of filoviruses. Arch Virol Suppl 7:81–100
Peters CJ, Sanchez A, Rollin PE, Ksiazek TG, Murphy FA (1996) Filoviruses. Fields virology. Lippincott-Raven, Philadelphia, PA
Schnell MJ, Buonocore L, Kretzschmar E, Johnson E, Rose JK (1996) Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles. Proc Natl Acad Sci U S A 93(21):11359–11365
Takada A, Robison C, Goto H, Sanchez A, Murti KG, Whitt MA, Kawaoka Y (1997) A system for functional analysis of Ebola virus glycoprotein. Proc Natl Acad Sci U S A 94(26):14764–14769
Ito H, Watanabe S, Sanchez A, Whitt MA, Kawaoka Y (1999) Mutational analysis of the putative fusion domain of Ebola virus glycoprotein. J Virol 73(10):8907–8912
Ito H, Watanabe S, Takada A, Kawaoka Y (2001) Ebola virus glycoprotein: proteolytic processing, acylation, cell tropism, and detection of neutralizing antibodies. J Virol 75(3):1576–1580. doi:10.1128/jvi.75.3.1576-1580.2001
Zavada J (1982) The pseudotypic paradox. J Gen Virol 63(Pt 1):15–24. doi:10.1099/0022-1317-63-1-15
Suda Y, Fukushi S, Tani H, Murakami S, Saijo M, Horimoto T, Shimojima M (2016) Analysis of the entry mechanism of Crimean-Congo hemorrhagic fever virus, using a vesicular stomatitis virus pseudotyping system. Arch Virol. doi:10.1007/s00705-016-2803-1
Shtanko O, Nikitina RA, Altuntas CZ, Chepurnov AA, Davey RA (2014) Crimean-Congo hemorrhagic fever virus entry into host cells occurs through the multivesicular body and requires ESCRT regulators. PLoS Pathog 10(9):e1004390. doi:10.1371/journal.ppat.1004390
Fukushi S, Tani H, Yoshikawa T, Saijo M, Morikawa S (2012) Serological assays based on recombinant viral proteins for the diagnosis of arenavirus hemorrhagic fevers. Virus 4(10):2097–2114. doi:10.3390/v4102097
Ray N, Whidby J, Stewart S, Hooper JW, Bertolotti-Ciarlet A (2010) Study of Andes virus entry and neutralization using a pseudovirion system. J Virol Methods 163(2):416–423. doi:10.1016/j.jviromet.2009.11.004
Moller-Tank S, Kondratowicz AS, Davey RA, Rennert PD, Maury W (2013) Role of the phosphatidylserine receptor TIM-1 in enveloped-virus entry. J Virol 87(15):8327–8341. doi:10.1128/JVI.01025-13
Tamin A, Harcourt BH, Lo MK, Roth JA, Wolf MC, Lee B, Weingartl H, Audonnet JC, Bellini WJ, Rota PA (2009) Development of a neutralization assay for Nipah virus using pseudotype particles. J Virol Methods 160(1–2):1–6. doi:10.1016/j.jviromet.2009.02.025
Ogino M, Ebihara H, Lee BH, Araki K, Lundkvist A, Kawaoka Y, Yoshimatsu K, Arikawa J (2003) Use of vesicular stomatitis virus pseudotypes bearing hantaan or seoul virus envelope proteins in a rapid and safe neutralization test. Clin Diagn Lab Immunol 10(1):154–160
Tani H, Iha K, Shimojima M, Fukushi S, Taniguchi S, Yoshikawa T, Kawaoka Y, Nakasone N, Ninomiya H, Saijo M, Morikawa S (2014) Analysis of Lujo virus cell entry using pseudotype vesicular stomatitis virus. J Virol 88(13):7317–7330. doi:10.1128/JVI.00512-14
Lawson ND, Stillman EA, Whitt MA, Rose JK (1995) Recombinant vesicular stomatitis viruses from DNA. Proc Natl Acad Sci U S A 92(10):4477–4481
Acknowledgments
This work was supported by the National Institutes of Health R21 AI123616 (W.M.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Brouillette, R.B., Maury, W. (2017). Production of Filovirus Glycoprotein-Pseudotyped Vesicular Stomatitis Virus for Study of Filovirus Entry Mechanisms. In: Hoenen, T., Groseth, A. (eds) Ebolaviruses. Methods in Molecular Biology, vol 1628. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7116-9_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7116-9_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7115-2
Online ISBN: 978-1-4939-7116-9
eBook Packages: Springer Protocols