Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection

doi:10.1016/S2095-3119(19)62717-6

Journal of Integrative Agriculture ›› 2019, Vol. 18 ›› Issue (10): 2361-2368.DOI: 10.1016/S2095-3119(19)62717-6

收稿日期:2019-01-14 出版日期:2019-10-01 发布日期:2019-09-29

Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection

ZHAO Dan-dan^1*, SHUAI Lei^1*, GE Jin-ying¹, WANG Jin-liang¹, WEN Zhi-yuan¹, LIU Ren-qiang¹, WANG Chong¹, WANG Xi-jun¹, BU Zhi-gao^{1, 2}

¹State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China
² Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, P.R.China

Received:2019-01-14 Online:2019-10-01 Published:2019-09-29
Contact: Correspondence BU Zhi-gao, E-mail: buzhigao@caas.cn; WANG Xi-jun, E-mail: wangxijun@caas.cn
About author:ZHAO Dan-dan, E-mail: 13633636807@163.com; SHUAI Lei, E-mail: shuailei@caas.cn; * These authors contributed equally to this study.
Supported by:
This work was supported by the National Natural Science Fundation of China (31800138) and the National Key Research and Development Program of China (2016YFD0500403).

摘要/Abstract

Abstract:

The mechanism of rabies virus (RABV) infection still needs to be further characterized. RABV particle with self-fluorescent is a powerful viral model to visualize the viral infection process in cells. Herein, based on a reverse genetic system of the Evelyn-Rokitnicki-Abelseth (rERA) strain, we generated a recombinant RABV rERA-N/mCherry strain that stably expresses an additional ERA nucleoprotein that fuses with the red fluorescent protein mCherry (N/mCherry). The rERA-N/mCherry strain retained growth property similar to the parent strain rERA in vitro. The N/mCherry expression showed genetic stability during passage into mouse neuroblastoma (NA) cells and did not change the virulence of the vector. The rERA-N/mCherry strain was then utilized as a visual viral model to study the RABV-cell binding and internalization. We directly observed the red self-fluorescence of rERA-N/mCherry particles binding to the cell surface, and further co-localizing with clathrin in the early stage of infection in NA cells by fluorescence microscopy. Our results showed that the rERA-N/mCherry strain uses clathrin-dependent endocytosis to enter cells, which is consistent with the well-known mechanism of RABV invasion. The recombinant RABV rERA-N/mCherry thus appears to have the potential to be an effective viral model to further explore the fundamental molecular mechanism of rabies neuropathogenesis.

Key words: rabies virus , self-fluorescence , binding , internalization

. [J]. Journal of Integrative Agriculture, 2019, 18(10): 2361-2368.

ZHAO Dan-dan, SHUAI Lei, GE Jin-ying, WANG Jin-liang, WEN Zhi-yuan, LIU Ren-qiang, WANG Chong, WANG Xi-jun, BU Zhi-gao. Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection[J]. Journal of Integrative Agriculture, 2019, 18(10): 2361-2368.

参考文献

Albertini A A, Schoehn G, Weissenhorn W, Ruigrok R W. 2008. Structural aspects of rabies virus replication. Cellular and Molecular Life Sciences, 65, 282–294.
Betzig E, Patterson G H, Sougrat R, Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincott-Schwartz J, Hess H F. 2006. Imaging intracellular fluorescent proteins at nanometer resolution. Science, 313, 1642–1645.
Boulant S, Stanifer M, Lozach P Y. 2015. Dynamics of virus-receptor interactions in virus binding, signaling, and endocytosis. Viruses, 7, 2794–2815.
Brandenburg B, Zhuang X. 2007. Virus trafficking - learning from single-virus tracking. Nature Reviews Microbiology, 5, 197–208.
Burckhardt C J, Greber U F. 2009. Virus movements on the plasma membrane support infection and transmission between cells. PLoS Pathogens, 5, e1000621.
Carette J E, Raaben M, Wong A C, Herbert A S, Obernosterer G, Mulherkar N, Kuehne A I, Kranzusch P J, Griffin A M, Ruthel G, Dal Cin P, Dye J M, Whelan S P, Chandran K, Brummelkamp T R. 2011. Ebola virus entry requires the cholesterol transporter Niemann-Pick C1. Nature, 477, 340–343.
Davis B M, Rall G F, Schnell M J. 2015. Everything you always wanted to know about rabies virus (but were afraid to ask). Annual Review of Virology, 2, 451.
Gaudin Y. 2000. Rabies virus-induced membrane fusion pathway. The Journal of Cell Biology, 150, 601–612.
Ge J, Wang X, Tao L, Wen Z, Feng N, Yang S, Xia X, Yang C, Chen H, Bu Z. 2011. Newcastle disease virus-vectored rabies vaccine is safe, highly immunogenic, and provides long-lasting protection in dogs and cats. Journal of Virology, 85, 8241–8252.
Greber U F, Way M. 2006. A superhighway to virus infection. Cell, 124, 741–754.
Helmuth J A, Burckhardt C J, Koumoutsakos P, Greber U F, Sbalzarini I F. 2007. A novel supervised trajectory segmentation algorithm identifies distinct types of human adenovirus motion in host cells. Journal of Structural Biology, 159, 347–358.
Jackson A C W W. 2007. Rabies. 2nd ed. Elsevier, Academic Press, Amsterdam, Boston.
Lawson K F, Walker V C, Crawley J F. 1967. ERA strain rabies vaccine. Duration of immunity in cattle, dogs and cats. Veterinary Medicine, Small Animal Clinician, 62, 1073–1074.
Lentz T L, Burrage T G, Smith A L, Crick J, Tignor G H. 1982. Is the acetylcholine receptor a rabies virus receptor? Science, 215, 182–184.
Lewis P, Lentz T L. 1998. Rabies virus entry into cultured rat hippocampal neurons. Journal of Neurocytology, 27, 559–573.
Mahadevan A, Suja M S, Mani R S, Shankar S K. 2016. Perspectives in diagnosis and treatment of rabies viral encephalitis: Insights from pathogenesis. Neurotherapeutics: The Journal of the American Society for Experimental Neuro Therapeutics, 13, 477–492.
Minghui R, Stone M, Semedo M H, Nel L. 2018. New global strategic plan to eliminate dog-mediated rabies by 2030. The Lancet Global Health, 6, e828–e829.
Mothes W, Sherer N M, Jin J, Zhong P. 2010. Virus Cell-to-Cell Transmission. Journal of Virology, 84, 8360–8368.
Piccinotti S, Kirchhausen T, Whelan S P. 2013. Uptake of rabies virus into epithelial cells by clathrin-mediated endocytosis depends upon actin. Journal of Virology, 87, 11637–11647.
Piccinotti S, Whelan S P. 2016. Rabies internalizes into primary peripheral neurons via clathrin coated pits and requires fusion at the cell body. PLoS Pathogens, 12, e1005753.
Roche S, Gaudin Y. 2004. Evidence that rabies virus forms different kinds of fusion machines with different pH thresholds for fusion. Journal of Virology, 78, 8746–8752.
Rupprecht C, Kuzmin I, Meslin F. 2017. Lyssaviruses and rabies: Current conundrums, concerns, contradictions and controversies. F1000research, 6, 184.
van der Schaar H M, Rust M J, Chen C, van der Ende-Metselaar H, Wilschut J, Zhuang X W, Smit J M. 2008. Dissecting the cell entry pathway of dengue virus by single-particle tracking in living cells. PLoS Pathogens, 4, e1000244.
Shuai L, Feng N, Wang X, Ge J, Wen Z, Chen W, Qin L, Xia X, Bu Z. 2015. Genetically modified rabies virus ERA strain is safe and induces long-lasting protective immune response in dogs after oral vaccination. Antiviral Research, 121, 9–15.
Shuai L, Wang X, Wen Z, Ge J, Wang J, Zhao D, Bu Z. 2017. Genetically modified rabies virus-vectored Ebola virus disease vaccines are safe and induce efficacious immune responses in mice and dogs. Antiviral Research, 146, 36–44.
Sun E, He J, Zhuang X. 2013. Live cell imaging of viral entry. Current Opinion in Virology, 3, 34–43.
Takeuchi M, Ozawa T. 2007. Methods for imaging and analyses of intracellular organelles using fluorescent and luminescent proteins. Analytical sciences: The International Journal of the Japan Society for Analytical Chemistry, 23, 25–29.
Tao L, Ge J, Wang X, Wen Z, Zhai H, Tao H, Zhao B, Kong D, Yang C, Bu Z. 2011. Generation of a recombinant rabies Flury LEP virus carrying an additional G gene creates an improved seed virus for inactivated vaccine production. Virology Journal, 8, 454–454.
Tao L, Ge J, Wang X, Zhai H, Hua T, Zhao B, Kong D, Yang C, Chen H, Bu Z. 2010. Molecular basis of neurovirulence of flury rabies virus vaccine strains: Importance of the polymerase and the glycoprotein R333Q mutation. Journal of Virology, 84, 8926–8936.
Thoulouze M I, Lafage M, Schachner M, Hartmann U, Cremer H, Lafon M. 1998. The neural cell adhesion molecule is a receptor for rabies virus. Journal of Virology, 72, 7181–7190.
Tuffereau C, Benejean J, Blondel D, Kieffer B, Flamand A. 1998. Low-affinity nerve-growth factor receptor (P75NTR) can serve as a receptor for rabies virus. EMBO Journal, 17, 7250–7259.
Wang J L, Wang Z L, Liu R Q, Shuai L, Wang X X, Luo J, Wang C, Chen W Y, Wang X J, Ge J Y, He X J, Wen Z Y, Bu Z G. 2018. Metabotropic glutamate receptor subtype 2 is a cellular receptor for rabies virus. PLoS Pathogens, 14, e1007189.
Wang X, Feng N, Ge J, Shuai L, Peng L, Gao Y, Yang S, Xia X, Bu Z. 2012. Recombinant canine distemper virus serves as bivalent live vaccine against rabies and canine distemper. Vaccine, 30, 5067–5072.
Worlein J M, Baker K, Bloomsmith M, Coleman K, Koban T L. 2011. The eighth edition of the guide for the care and use of laboratory animals (2011): Implications for behavioral management. American Journal of Primatology, 73, 98–98.
Xu H J, Hao X, Wang S W, Wang Z Y, Cai M J, Jiang J G, Qin Q W, Zhang M L, Wang H D. 2015. Real-time imaging of rabies virus entry into living vero cells. Scientifc Reports, 5, 11753.

Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection

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