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Mutation of I176R in the E coding region weakens Japanese encephalitis virus neurovirulence, but not its growth rate in BHK-21 cells

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Abstract

Previously, we isolated the Japanese encephalitis virus (JEV) strain SCYA201201. In this study, we passed the SCYA201201 strain in Syrian baby hamster kidney (BHK-21) cells 120 times to obtain the SCYA201201-0901 strain, which exhibited an attenuated phenotype in mice. Comparison of SCYA201201-0901 amino acid sequences with those of other JEV strains revealed a single mutation, I176R, in the E coding region. Using reverse genetic technology, we provide evidence that this single E-I176R mutation does not affect virus growth in BHK-21 cells but significantly decreases JEV neurovirulence in mice. This study provides critical information for understanding the molecular mechanism of JEV attenuation.

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References

  1. Xin YY, Ming ZG, Peng GY, Jian A, Min LH (1988) Safety of a live-attenuated Japanese encephalitis virus vaccine (SA14-14-2) for children. Am J Trop Med Hyg 39(2):214–217

    Article  CAS  PubMed  Google Scholar 

  2. Yun SI, Song BH, Polejaeva IA, Davies CJ, White KL, Lee YM (2016) Comparison of the live-attenuated Japanese encephalitis vaccine SA14-14-2 strain with its pre-attenuated virulent parent SA14 strain: similarities and differences in vitro and in vivo. J Gen Virol 97(10):2575

    Article  CAS  PubMed  Google Scholar 

  3. Ni H, Burns NJ, Chang GJ, Zhang MJ, Wills MR, Trent DW, Sanders PG, Barrett AD (1994) Comparison of nucleotide and deduced amino acid sequence of the 5′ non-coding region and structural protein genes of the wild-type Japanese encephalitis virus strain SA14 and its attenuated vaccine derivatives. J Gen Virol 75(Pt 6):1505

    Article  CAS  PubMed  Google Scholar 

  4. Zhao Z, Date T, Li Y, Kato T, Miyamoto M, Yasui K, Wakita T (2005) Characterization of the E-138 (Glu/Lys) mutation in Japanese encephalitis virus by using a stable, full-length, infectious cDNA clone. J Gen Virol 86(8):2209–2220

    Article  CAS  PubMed  Google Scholar 

  5. Gromowski GD, Firestone CY, Whitehead SS (2015) Genetic determinants of Japanese encephalitis virus vaccine strain SA14-14-2 that govern attenuation of virulence in mice. J Virol 89(12):6328–6337

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Tajima S, Nerome R, Nukui Y, Kato F, Takasaki T, Kurane I (2010) A single mutation in the Japanese encephalitis virus E protein (S123R) increases its growth rate in mouse neuroblastoma cells and its pathogenicity in mice. Virology 396(2):298–304

    Article  CAS  PubMed  Google Scholar 

  7. Lindenbach BD, Thiel HJ, Rice CM (2007) Flaviviridae: the viruses and their replication. In: Knipe DM, Howley PM, Griffin DE, Lamb RA, Martin MA, Roizman B, Straus SE (eds) Fields virology, 5th edn. Lippincott Williams & Wilkins, Philadelphia, pp 1101–1152

    Google Scholar 

  8. Yu YX (2010) Phenotypic and genotypic characteristics of Japanese encephalitis attenuated live vaccine virus SA14-14-2 and their stabilities. Vaccine 28(21):3635–3641

    Article  CAS  PubMed  Google Scholar 

  9. Yang J, Yang H, Li Z, Wang W, Lin H, Liu L, Ni Q, Liu X, Zeng X, Wu Y (2017) Envelope protein mutations L107F and E138K are important for neurovirulence attenuation for japanese encephalitis virus SA14-14-2 strain. Viruses 9(1):20

    Article  PubMed Central  Google Scholar 

  10. Arroyo J, Guirakhoo F, Fenner S, Zhang ZX, Monath TP, Chambers TJ (2001) Molecular basis for attenuation of neurovirulence of a yellow fever virus/japanese encephalitis virus chimera vaccine (ChimeriVax-JE). J Virol 75(2):934–942

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Yuan L, Wu R, Liu H, Wen X, Huang X, Wen Y, Ma X, Yan Q, Huang Y, Zhao Q (2016) Tissue tropism and molecular characterization of a Japanese encephalitis virus strains isolated from pigs in southwest China. Virus Res 215:55–64

    Article  CAS  PubMed  Google Scholar 

  12. Solomon T, Ni H, Beasley DW, Ekkelenkamp M, Cardosa MJ, Barrett AD (2003) Origin and evolution of Japanese encephalitis virus in southeast Asia. J Virol 77(5):3091–3098

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Pujhari SK, Prabhakar S, Ratho RK, Modi M, Sharma M, Mishra B (2011) A novel mutation (S227T) in domain II of the envelope gene of Japanese encephalitis virus circulating in North India. Epidemiol Infect 139(6):849–856

    Article  CAS  PubMed  Google Scholar 

  14. Lee E, Lobigs M (2000) Substitutions at the putative receptor-binding site of an encephalitic flavivirus alter virulence and host cell tropism and reveal a role for glycosaminoglycans in entry. J Virol 74(19):8867–8875

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Chen Y, Maguire T, Hileman RE, Fromm JR, Esko JD, Linhardt RJ, Marks RM (1997) Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nat Med 3(8):866–871

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the National Program on Key Research Project of China (2016YFD0500403).

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Author notes

  1. Yuyong Zhou and Rui Wu, the authors contributed equally to this study.

Authors and Affiliations

  1. Research Center of Swine Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China

    Yuyong Zhou, Rui Wu, Qin Zhao, Xintian Wen, Yao Feng, Xiaobo Huang, Yiping Wen, Qigui Yan, Yong Huang, Xiaoping Ma, Xinfeng Han & Sanjie Cao

  2. Sichuan Science-Observation Experiment of Veterinary Drugs and Veterinary Diagnostic Technology, Ministry of Agriculture, Chengdu, 611130, China

    Yuyong Zhou, Rui Wu, Qin Zhao, Xintian Wen, Yao Feng, Xiaobo Huang, Yiping Wen, Qigui Yan, Yong Huang, Xiaoping Ma, Xinfeng Han & Sanjie Cao

  3. Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA

    Yung-Fu Chang

  4. National Teaching and Experiment Center of Animal, Sichuan Agricultural University, Chengdu, 611130, China

    Qin Zhao & Sanjie Cao

Authors
  1. Yuyong Zhou
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  2. Rui Wu
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  3. Qin Zhao
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  4. Yung-Fu Chang
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  5. Xintian Wen
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  6. Yao Feng
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  7. Xiaobo Huang
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  8. Yiping Wen
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  9. Qigui Yan
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  10. Yong Huang
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  11. Xiaoping Ma
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  12. Xinfeng Han
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  13. Sanjie Cao
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Corresponding author

Correspondence to Sanjie Cao.

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Conflict of interest

The authors declare that there is no conflict of interest.

Ethical statement

All animal experiments were approved by the Committee of the Ethics on Animal Care and Experiments at Sichuan Agricultural University. All experimental procedures were carried out in accordance with the approved guidelines.

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Handling Editor: Sheela Ramamoorthy.

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Zhou, Y., Wu, R., Zhao, Q. et al. Mutation of I176R in the E coding region weakens Japanese encephalitis virus neurovirulence, but not its growth rate in BHK-21 cells. Arch Virol 163, 1351–1355 (2018). https://doi.org/10.1007/s00705-018-3765-2

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  • DOI: https://doi.org/10.1007/s00705-018-3765-2

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