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Rescue and characterization of a recombinant HY12 bovine enterovirus carrying a foreign HA epitope in the 3A nonstructural protein

  • Dan Liu
  • Changming Liu
  • Xing Liu
  • Xin Li
  • Liping Huang
  • Junying Hu
  • Yanwu Wei
  • Hongzhen Zhu
  • Qun Zhang
  • Xinping WangEmail author
Original Article
  • 35 Downloads

Abstract

Full-length infectious cDNA clones for recombinant HY12 bovine enteroviruses designated as rHY12-3A-2-HA, rHY12-3A-3-HA, and rHY12-3A-9-HA were constructed by the insertion of an epitope from influenza virus hemagglutinin (HA) at the N-terminus of the HY12-encoded 3A protein at amino acid positions 2, 3, and 9. The recombinant HY12 viruses expressing the HA epitope were rescued and characterized using immunoperoxidase monolayer assay, western blotting, and electron microscopy. The three rescued recombinant marker viruses showed similar characteristics, such as TCID50 titer, plaque size, and growth properties, to those of parental rHY12 virus. Comparative analysis of the nucleotide sequences demonstrated the three recombinant marker viruses remained stable for 15 passages with no genetic changes. The recombinant viruses remained viable in various permissive cell lines, including BHK-21, Vero, and PK15 cells, suggesting that the insertion of the HA epitope tag had no effect on virus infectivity. Mice infected with the recombinant marker viruses and the parental virus produced anti-HY12-virus antibodies, while the recombinant marker viruses also produced anti-HA-epitope-tag antibodies. Taken together, these results demonstrate that HY12 viruses containing genetic markers may be useful tools for future investigations of the mechanisms of viral pathogenesis and virus replication, as well as for vaccine development.

Notes

Author contributions

We thank Dr. Yanjin Zhang at the University of Maryland for providing us a gift of the pCI-T7 plasmid. We also thank Dr. Encheng Sun, Dr. Liyan Cao at the Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, for technical assistance and helpful suggestions.

Funding

This work was supported by grants from the National Natural Science Foundation of China (31572531), National Key Research and Development Programs (2017YFD0500104, 2016YFD0500904, and 2017YFD0500903) and the Natural Science Foundation of Heilongjiang Province in China (C2015064).

Compliance with ethical standards

Conflicts of interest

The authors declare that there are no conflicts of interest.

Ethical approval

The handling of mice and the procedures used for this study were done following a standard protocol reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Jilin University (approval no JLU-20150226), in strict compliance with the requirements of the Animal Ethics Procedures and Guidelines of the People’s Republic of China.

References

  1. 1.
    Barya MA, Moll T, Mattson DE (1967) Antigenic analysis of bovine enteroviruses through studies of the kinetics of neutralization. Am J Vet Res 28:1283–1294Google Scholar
  2. 2.
    Chu JQ, Lee YJ, Park JN, Kim SM, Lee KN, Ko YJ, Lee HS, Cho IS, Kim B, Park JH (2013) Construction of a bovine enterovirus-based vector expressing a foot-and-mouth disease virus epitope. J Virol Methods 189:101–104CrossRefGoogle Scholar
  3. 3.
    Datta U, Dasgupta A (1994) Expression and subcellular localization of poliovirus VPg-precursor protein 3AB in eukaryotic cells: evidence for glycosylation in vitro. J Virol 68:4468–4477Google Scholar
  4. 4.
    Dunne HW, Huang CM, Lin WJ (1974) Bovine enteroviruses in the calf: an attempt at serologic, biologic, and pathologic classification. J Am Vet Med Assoc 164:290–294Google Scholar
  5. 5.
    Forss S, Strebel K, Beck E, Schaller H (1984) Nucleotide sequence and genome organization of foot-and-mouth disease virus. Nucl Acids Res 12:6587–6601CrossRefGoogle Scholar
  6. 6.
    Fujita K, Krishnakumar SS, Franco D, Paul AV, London E, Wimmer E (2007) Membrane topography of the hydrophobic anchor sequence of poliovirus 3A and 3AB proteins and the functional effect of 3A/3AB membrane association upon RNA replication. Biochemistry 46:5185–5199CrossRefGoogle Scholar
  7. 7.
    Gai X, Zhang Q, Lu H, Yang Z, Zhu L, Li X, Wang X (2018) A neonatal murine model for evaluation of enterovirus E HY12 virus infection and pathogenicity. PLoS One 13:e0193155CrossRefGoogle Scholar
  8. 8.
    Giachetti C, Hwang SS, Semler BL (1992) cis-acting lesions targeted to the hydrophobic domain of a poliovirus membrane protein involved in RNA replication. J Virol 66:6045–6057Google Scholar
  9. 9.
    Gur S, Yapkic O, Yilmaz A (2008) Serological survey of bovine enterovirus type 1 in different mammalian species in Turkey. Zoonoses Public Hl 55:106–111CrossRefGoogle Scholar
  10. 10.
    Han Q, Williams WB, Saunders KO, Seaton KE, Wiehe KJ, Vandergrift N, Von Holle TA, Trama AM, Parks RJ, Luo K, Gurley TC, Kepler TB, Marshall DJ, Montefiori DC, Sutherland LL, Alam MS, Whitesides JF, Bowman CM, Permar SR, Graham BS, Mascola JR, Seed PC, Van Rompay KKA, Tomaras GD, Moody MA, Haynes BF (2017) HIV DNA-adenovirus multiclade envelope vaccine induces gp41 Antibody immunodominance in rhesus macaques. J Virol 91(21):e00923–17CrossRefGoogle Scholar
  11. 11.
    Giraudo AT, Beck E, Strebel K, de Mello PA, La Torre JL, Scodeller EA, Bergmann IE (1990) Identification of a nucleotide deletion in parts of polypeptide 3A in two independent attenuated aphthovirus strains. Virology 177:780–783CrossRefGoogle Scholar
  12. 12.
    Jiang H, Weng L, Zhang N, Arita M, Li R, Chen L, Toyoda T (2011) Biochemical characterization of enterovirus 71 3D RNA polymerase. Biochim Biophys Acta 1809:211–219CrossRefGoogle Scholar
  13. 13.
    Knox C, Moffat K, Ali S, Ryan M, Wileman T (2005) Foot-and-mouth disease virus replication sites form next to the nucleus and close to the Golgi apparatus, but exclude marker proteins associated with host membrane compartments. J Gen Virol 86:687–696CrossRefGoogle Scholar
  14. 14.
    Kunin CM, Minuse E (1958) The isolation in tissue culture, chick embryo and suckling mice of filtrable agents from healthy dairy cattle. J Immunol 80:1–11Google Scholar
  15. 15.
    Lama J, Sanz MA, Carrasco L (1998) Genetic analysis of poliovirus protein 3A: characterization of a non-cytopathic mutant virus defective in killing Vero cells. J Gen Virol 79(Pt 8):1911–1921CrossRefGoogle Scholar
  16. 16.
    Li P, Bai X, Cao Y, Han C, Lu Z, Sun P, Yin H, Liu Z (2012) Expression and stability of foreign epitopes introduced into 3A nonstructural protein of foot-and-mouth disease virus. PLoS One 7:e41486CrossRefGoogle Scholar
  17. 17.
    Li Y, Chang J, Wang Q, Yu L (2012) Isolation of two Chinese bovine enteroviruses and sequence analysis of their complete genomes. Arch Virol 157:2369–2375CrossRefGoogle Scholar
  18. 18.
    Liu C, Ihara T, Nunoya T, Ueda S (2004) Development of an ELISA based on the baculovirus-expressed capsid protein of porcine circovirus type 2 as antigen. J Vet Med Sci 66:237–242CrossRefGoogle Scholar
  19. 19.
    Liu D, Liu C, Hu J, Hang L, Li X, Wei Y, Zhu H, Zhang Q, Wang X (2018) Construction and evaluation of HA-epitope-tag introduction onto the VP1 structural protein of a novel HY12 enterovirus. Virology 525:106–116CrossRefGoogle Scholar
  20. 20.
    Lu HH, Li X, Cuconati A, Wimmer E (1995) Analysis of picornavirus 2A(pro) proteins: separation of proteinase from translation and replication functions. J Virol 69:7445–7452Google Scholar
  21. 21.
    Ma X, Li P, Sun P, Bai X, Bao H, Lu Z, Fu Y, Cao Y, Li D, Chen Y, Qiao Z, Liu Z (2015) Construction and characterization of 3A-epitope-tagged foot-and-mouth disease virus. Infect Genet Evol 31:17–24CrossRefGoogle Scholar
  22. 22.
    Moll T, Davis AD (1959) Isolation and characterization of cytopathogenic enteroviruses from cattle with respiratory disease. Am J Vet Res 20:27–32Google Scholar
  23. 23.
    O’Donnell VK, Pacheco JM, Henry TM, Mason PW (2001) Subcellular distribution of the foot-and-mouth disease virus 3A protein in cells infected with viruses encoding wild-type and bovine-attenuated forms of 3A. Virology 287:151–162CrossRefGoogle Scholar
  24. 24.
    Paul AV (2002) Possible unifying mechanism of picornavirus genome replication. In: Semler BL (ed) In Molecular Biology of PicornaViruses. ASM Press, Washington, DC, pp 227–246Google Scholar
  25. 25.
    Sena-Esteves M, Gao G (2018) Production of high-titer retrovirus and lentivirus vectors. Cold Spring Harb Protoc 2018(4):pdb.prot095687CrossRefGoogle Scholar
  26. 26.
    Shang B, Deng C, Ye H, Xu W, Yuan Z, Shi PY, Zhang B (2013) Development and characterization of a stable eGFP enterovirus 71 for antiviral screening. Antivir Res 97:198–205CrossRefGoogle Scholar
  27. 27.
    Shingu M, Chinami M, Taguchi T, Shingu M Jr (1991) Therapeutic effects of bovine enterovirus infection on rabbits with experimentally induced adult T cell leukaemia. J Gen Virol 72(Pt 8):2031–2034CrossRefGoogle Scholar
  28. 28.
    Tang WF, Yang SY, Wu BW, Jheng JR, Chen YL, Shih CH, Lin KH, Lai HC, Tang P, Horng JT (2007) Reticulon 3 binds the 2C protein of enterovirus 71 and is required for viral replication. J Biol Chem 282:5888–5898CrossRefGoogle Scholar
  29. 29.
    Teterina NL, Levenson EA, Ehrenfeld E (2010) Viable polioviruses that encode 2A proteins with fluorescent protein tags. J Virol 84:1477–1488CrossRefGoogle Scholar
  30. 30.
    Teterina NL, Lauber C, Jensen KS, Levenson EA, Gorbalenya AE, Ehrenfeld E (2011) Identification of tolerated insertion sites in poliovirus non-structural proteins. Virology 409:1–11CrossRefGoogle Scholar
  31. 31.
    Teterina NL, Pinto Y, Weaver JD, Jensen KS, Ehrenfeld E (2011) Analysis of poliovirus protein 3A interactions with viral and cellular proteins in infected cells. J Virol 85:4284–4296CrossRefGoogle Scholar
  32. 32.
    Towner JS, Ho TV, Semler BL (1996) Determinants of membrane association for poliovirus protein 3AB. J Biol Chem 271:26810–26818CrossRefGoogle Scholar
  33. 33.
    Towner JS, Brown DM, Nguyen JH, Semler BL (2003) Functional conservation of the hydrophobic domain of polypeptide 3AB between human rhinovirus and poliovirus. Virology 314:432–442CrossRefGoogle Scholar
  34. 34.
    Tsuchiaka S, Rahpaya SS, Otomaru K, Aoki H, Kishimoto M, Naoi Y, Omatsu T, Sano K, Okazaki-Terashima S, Katayama Y, Oba M, Nagai M, Mizutani T (2017) Identification of a novel bovine enterovirus possessing highly divergent amino acid sequences in capsid protein. BMC Microbiol 17:18CrossRefGoogle Scholar
  35. 35.
    Wang M, He J, Lu H, Liu Y, Deng Y, Zhu L, Guo C, Tu C, Wang X (2017) A novel enterovirus species identified from severe diarrheal goats. PLoS One 12:e0174600CrossRefGoogle Scholar
  36. 36.
    Zhu L, Xing Z, Gai X, Li S, San Z, Wang X (2014) Identification of a novel enterovirus E isolates HY12 from cattle with severe respiratory and enteric diseases. PLoS One 9:e97730CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Dan Liu
    • 1
  • Changming Liu
    • 2
  • Xing Liu
    • 3
  • Xin Li
    • 1
  • Liping Huang
    • 2
  • Junying Hu
    • 1
  • Yanwu Wei
    • 2
  • Hongzhen Zhu
    • 2
  • Qun Zhang
    • 1
  • Xinping Wang
    • 1
    Email author
  1. 1.Key Laboratory for Zoonoses Research, Ministry of Education, College of Veterinary MedicineJilin UniversityChangchunChina
  2. 2.State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbinChina
  3. 3.Inactivated Vaccine Production Workshop Comprehensive GroupHarbin Weike Biotechnology Development CompanyHarbinChina

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