Skip to main content
SpringerLink
Log in

Expression of chimeric proteins based on a backbone of the GII.4 norovirus VP1 and their application in the study of a GII.6 norovirus-specific blockade epitope

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

The surface-exposed loop regions of the protruding domain of the norovirus (NoV) major capsid protein VP1 can tolerate the insertion of foreign antigens without affecting its assembly into subviral particles. In this study, we investigated the tolerance of the surface-exposed loop region of the GII.4 NoV VP1 by replacing it with homologous or heterologous sequences. We designed a panel of constructs in which the amino acid sequence from position 298-305 of the GII.4 NoV VP1 was replaced by sequences derived from the same region of GI.3, GII.3, GII.6, and GII.17 NoVs as well as neutralizing epitopes of enterovirus type 71 and varicella-zoster virus. The constructs were synthesized and expressed using a recombinant baculovirus expression system. The expression of target proteins was measured by indirect enzyme-linked immunosorbent assay (ELISA), and the assembly of virus-like particles (VLPs) was confirmed by electron microscopy. Our results showed that all of the constructs expressed high levels of target chimeric proteins, and all of the chimeric proteins successfully assembled into VLPs or subviral particles. An in vitro VLP-histo-blood group antigen (HBGA) binding assay revealed that chimeric-protein-containing VLPs did not bind or showed reduced binding to salivary HBGAs, a ligand for NoV particles. The results of an in vitro VLP-HBGA binding blockade assay indicated that the predicted surface-exposed loop region of the GII.6 NoV VP1 may comprise a blockade epitope. In summary, the surface-exposed loop region of the GII.4 NoV VP1 can be replaced by foreign sequences of a certain length. Using this strategy, we found that the predicted surface-exposed loop region of GII.6 NoV VP1 might contain a blockade epitope.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Jiang X, Wang M, Graham DY, Estes MK (1992) Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J Virol 66(11):6527–6532

    Article  CAS  Google Scholar 

  2. Huo Y, Wan X, Ling T, Wu J, Wang W, Shen S (2018) Expression and purification of norovirus virus like particles in Escherichia coli and their immunogenicity in mice. Mol Immunol 93:278–284

    Article  CAS  Google Scholar 

  3. Prasad BV, Rothnagel R, Jiang X, Estes MK (1994) Three-dimensional structure of baculovirus-expressed Norwalk virus capsids. J Virol 68(8):5117–5125

    Article  CAS  Google Scholar 

  4. Prasad BV, Hardy ME, Dokland T, Bella J, Rossmann MG, Estes MK (1999) X-ray crystallographic structure of the Norwalk virus capsid. Science 286(5438):287–290

    Article  CAS  Google Scholar 

  5. Tan M, Fang P, Chachiyo T, Xia M, Huang P, Fang Z et al (2008) Noroviral P particle: structure, function and applications in virus-host interaction. Virology 382(1):115–123

    Article  CAS  Google Scholar 

  6. Tan M, Jiang X (2012) Norovirus P particle: a subviral nanoparticle for vaccine development against norovirus, rotavirus and influenza virus. Nanomedicine (Lond) 7(6):889–897

    Article  CAS  Google Scholar 

  7. Ming T, Ming X, Huang P, Wang L, Zhong W, Mcneal M et al (2011) Norovirus P particle as a platform for antigen presentation. Procedia Vaccinol 4:19–26

    Article  Google Scholar 

  8. Zheng L, Wang W, Liu J, Huo Y, Qin C, Wang M et al (2017) Comprehensive characterization of a major capsid protein derived from a documented GII.6 norovirus strain. Arch Virol 162(12):3863–3868

    Article  CAS  Google Scholar 

  9. Huo Y, Wang W, Zheng L, Chen X, Shen S, Wang M (2017) Enzymatic cleavage promotes disassembly of GII.3 norovirus virus like particles and its binding to salivary histo-blood group antigens. Virus Res 240:18–24

    Article  CAS  Google Scholar 

  10. Qiu S, Zheng L, Qin C, Yin X, Ma J, Liu J et al (2020) Production and characterization of monoclonal antibodies against GII.6 norovirus virus-like particles. Microb Pathog 142:104100

    Article  CAS  Google Scholar 

  11. Foo DG, Alonso S, Phoon MC, Ramachandran NP, Chow VT, Poh CL (2007) Identification of neutralizing linear epitopes from the VP1 capsid protein of Enterovirus 71 using synthetic peptides. Virus Res 125(1):61–68

    Article  CAS  Google Scholar 

  12. Zhu R, Liu J, Chen C, Ye X, Xu L, Wang W et al (2016) A highly conserved epitope-vaccine candidate against varicella-zoster virus induces neutralizing antibodies in mice. Vaccine 34(13):1589–1596

    Article  CAS  Google Scholar 

  13. Zhang G, Wang J, Liu J, Zheng L, Wang W, Huo Y et al (2019) The surface-exposed loop region of norovirus GII.3 VP1 plays an essential role in binding histo-blood group antigens. Arch Virol 164(6):1629–1638

    Article  CAS  Google Scholar 

  14. Zheng L, Wang W, Liu J, Chen X, Li S, Wang Q et al (2018) Characterization of a Norovirus-specific monoclonal antibody that exhibits wide spectrum binding activities. J Med Virol 90(4):671–676

    Article  CAS  Google Scholar 

  15. Huo Y, Wan X, Ling T, Shen S (2016) Biological and immunological characterization of norovirus major capsid proteins from three different genotypes. Microb Pathog 90:78–83

    Article  CAS  Google Scholar 

  16. Huo Y, Wan X, Wang Z, Meng S, Shen S (2015) Production of Norovirus VLPs to size homogeneity. Virus Res 204:1–5

    Article  CAS  Google Scholar 

  17. White LJ, Hardy ME, Estes MK (1997) Biochemical characterization of a smaller form of recombinant Norwalk virus capsids assembled in insect cells. J Virol 71(10):8066–8072

    Article  CAS  Google Scholar 

  18. Debbink K, Lindesmith LC, Donaldson EF, Swanstrom J, Baric RS (2014) Chimeric GII.4 norovirus virus-like-particle-based vaccines induce broadly blocking immune responses. J Virol 88(13):7256–7266

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank all the members of the laboratory for their contributions.

Funding

This research was supported by the Chinese Federation of Public Health Foundation (GWLM202003).

Author information

Authors and Affiliations

  1. The Sixth People’s Hospital of Zhengzhou, Zhengzhou, 450000, China

    Yuqi Huo, Jie Ma, Lijun Zheng, Jinjin Liu, Zhaojie Yang, Chao Wang & Qingxia Zhao

Authors
  1. Yuqi Huo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jie Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lijun Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jinjin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhaojie Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Qingxia Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YH and QZ conceived and designed the experiments; JM, LZ, JL, ZY, and CW performed the experiments. JM and LZ analyzed the data; YH prepared the manuscript.

Corresponding authors

Correspondence to Yuqi Huo or Qingxia Zhao.

Ethics declarations

Conflict of interest

The authors declare no conflicts of interest.

Ethical approval

Animal experiments were carried out at Wuhan Dia-an, following the guidelines of the Chinese Council on Animal Care.

Additional information

Handling Editor: Tim Skern.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PPTX 63 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huo, Y., Ma, J., Zheng, L. et al. Expression of chimeric proteins based on a backbone of the GII.4 norovirus VP1 and their application in the study of a GII.6 norovirus-specific blockade epitope. Arch Virol 167, 819–827 (2022). https://doi.org/10.1007/s00705-022-05362-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-022-05362-5

Search

Navigation