Virus-like particles based on rotavarus A recombinant VP2/VP6 proteins for assessment the antibody immune response by ELISA

Cover Page

Cite item

Full Text

Abstract

Introduction. Rotavirus infection is one of the main concerns in infectious pathology in humans, mammals and birds. Newborn piglets or rodents are usually being used as a laboratory model for the evaluation of immunogenicity and efficacy for all types of vaccines against rotavirus A (RVA), and the use of ELISA for the detection of virus-specific antibodies of specific isotype is an essential step of this evaluation.

Objective. Development of indirect solid-phase ELISA with VP2/VP6 rotavirus VLP as an antigen to detect and assess the distribution of RVA-specific IgG, IgM and IgA in the immune response to rotavirus A.

Materials and methods. VP2/VP6 rotavirus VLP production and purification, electron microscopy, PAGE, immunoblotting, ELISA, virus neutralization assay.

Results. The study presents the results of development of a recombinant baculovirus with RVA genes VP2-eGFP/VP6, assessment of its infectious activity and using it for VLP production. The morphology of the VP2/VP6 rotavirus VLPs was assessed, the structural composition was determined, and the high antigenic activity of the VLP was established. VLP-based ELISA assay was developed and here we report results for RVA-specific antibody detection in sera of different animals.

Conclusion. The developed ELISA based on VP2/VP6 rotavirus VLP as a universal antigen makes it possible to detect separately IgG, IgM and IgA antibodies to rotavirus A, outlining its scientific and practical importance for the evaluation of immunogenicity and efficacy of traditional vaccines against rotavirus A and those under development.

About the authors

Ilya E. Filatov

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Author for correspondence.
Email: filat69rus@yandex.ru
ORCID iD: 0000-0001-5274-224X

Junior Researcher, Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

Valery V. Tsibezov

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: tsibezov@yandex.ru
ORCID iD: 0000-0003-2150-5764

PhD, Leading Researcher

Russian Federation, 123098, Moscow

Marina V. Balandina

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: mbalandina77@mail.ru
ORCID iD: 0009-0002-8179-1379

Senior Researcher, Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

Svetlana N. Norkina

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: snork9@list.ru
ORCID iD: 0009-0006-9608-4713

Senior Researcher

Russian Federation, 123098, Moscow

Oleg E. Latyshev

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: oleglat80@mail.ru
ORCID iD: 0000-0002-5757-3809

PhD, senior researcher at the Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

Olesia V. Eliseeva

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: olesenka80@mail.ru
ORCID iD: 0000-0002-0723-9749

Senior Researcher at the Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

Stanislav A. Cherepushkin

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: cherepushkin1@gmail.com
ORCID iD: 0000-0002-1734-5369

Researcher, Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

Oleg A. Verkhovsky

Diagnostic and Prevention Research Institute for human and animal diseases

Email: info@dpri.com
ORCID iD: 0000-0003-0784-9341

Professor

Russian Federation, 123098, Moscow

Tatyana V. Grebennikova

National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya of the Ministry of Health of the Russian Federation

Email: t_grebennikova@mail.ru
ORCID iD: 0000-0002-6141-9361

Doctor of Biological Sciences, Professor, Corresponding Member RAS, Head Laboratory of Molecular Diagnostics

Russian Federation, 123098, Moscow

References

  1. L’vov D.K., ed. Virology Guide: Human and Animal Viruses and Viral Infections [Rukovodstvo po virusologii: Virusy i virusnye infektsii cheloveka i zhivotnykh]. Moscow: MIA; 2013. (in Russian)
  2. Saif L.J., Fernandez F.M. Group A rotavirus veterinary vaccines. J. Infect. Dis. 1996; 174(Suppl. 1): S98–106. https://doi.org/10.1093/infdis/174.supplement_1.s98
  3. Kumar D., Anderson A.V., Pittman J., Springer N.L., Marthaler D.G., Mwangi W. Antibody response to rotavirus C pre-farrow natural planned exposure to gilts and their piglets. Viruses. 2022; 14(10): 2250. https://doi.org/10.3390/v14102250
  4. Vetter V., Gardner R.C., Debrus S., Benninghoff B., Pereira P. Established and new rotavirus vaccines: a comprehensive review for healthcare professionals. Hum. Vaccin. Immunother. 2022; 18(1): 1870395. https://doi.org/10.1080/21645515.2020.1870395
  5. Folorunso O.S., Sebolai O.M. Overview of the development, impacts, and challenges of live-attenuated oral rotavirus vaccines. Vaccines (Basel). 2020; 8(3): 341. https://doi.org/10.3390/vaccines8030341
  6. Wang Y., Li J., Liu P., Zhu F. The performance of licensed rotavirus vaccines and the development of a new generation of rotavirus vaccines: a review. Hum. Vaccin. Immunother. 2021; 17(3): 880–96. https://doi.org/10.1080/21645515.2020.1801071
  7. Chepngeno J., Diaz A., Paim F.C., Saif L.J., Vlasova A.N. Rotavirus C: prevalence in suckling piglets and development of virus-like particles to assess the influence of maternal immunity on the disease development. Vet. Res. 2019; 50(1): 84. https://doi.org/10.1186/s13567-019-0705-4
  8. Changotra H., Vij A. Rotavirus virus-like particles (RV-VLPs) vaccines: An update. Rev. Med. Virol. 2017; 27(6). https://doi.org/10.1002/rmv.1954
  9. Jere K.C., O’Neill H.G., Potgieter A.C., van Dijk A.A. Chimaeric virus-like particles derived from consensus genome sequences of human rotavirus strains co-circulating in Africa. PLoS One. 2014; 9(9): e105167. https://doi.org/10.1371/journal.pone.0105167
  10. Song J.M. Parenteral, non-live rotavirus vaccine: recent history and future perspective. Clin. Exp. Vaccine Res. 2021; 10(3): 203–10. https://doi.org/10.7774/cevr.2021.10.3.203
  11. Crawford S.E., Estes M.K., Ciarlet M., Barone C., O’Neal C.M., Cohen J., et al. Heterotypic protection and induction of a broad heterotypic neutralization response by rotavirus-like particles. J. Virol. 1999; 73(6): 4813–22. https://doi.org/10.1128/jvi.73.6.4813-4822.1999
  12. Cherepushkin S.A., Tsibezov V.V., Yuzhakov A.G., Latyshev O.E., Alekseev K.P., Altaeva E.G., et al. Synthesis and characterization of human rotavirus A (Reoviridae: Sedoreovirinae: Rotavirus: Rotavirus A) virus-like particles. Voprosy virusologii. 2021; 66(1): 55–64. https://doi.org/10.36233/0507-4088-27 EDN: https://www.elibrary.ru/eersag (in Russian)
  13. Yuan L., Saif L.J. Induction of mucosal immune responses and protection against enteric viruses: rotavirus infection of gnotobiotic pigs as a model. Vet. Immunol. Immunopathol. 2002; 87(3–4): 147–60. https://doi.org/10.1016/s0165-2427(02)00046-6
  14. Azevedo M.P., Vlasova A.N., Saif L.J. Human rotavirus virus-like particle vaccines evaluated in a neonatal gnotobiotic pig model of human rotavirus disease. Expert. Rev. Vaccines. 2013; 12(2): 169–81. https://doi.org/10.1586/erv.13.3
  15. Shoja Z., Jalilvand S., Latifi T., Roohvand F. Rotavirus VP6: involvement in immunogenicity, adjuvant activity, and use as a vector for heterologous peptides, drug delivery, and production of nano-biomaterials. Arch. Virol. 2022; 167(4): 1013–23. https://doi.org/10.1007/s00705-022-05407-9
  16. Latyshev O.E., Eliseeva O.V., Kostina L.V., Alekseev K.P., Khametova K.M., Altaeva E.G., et al. Assessment of immunogenic activity of the cloned human rotavirus A WA strain. Voprosy virusologii. 2019; 64(4): 156–64. https://doi.org/10.36233/0507-4088-2019-64-4-156-164 EDN: https://www.elibrary.ru/sckbyy (in Russian)
  17. Lai C.C., Cheng Y.C., Chen P.W., Lin T.H., Tzeng T.T., Lu C.C., et al. Process development for pandemic influenza VLP vaccine production using a baculovirus expression system. J. Biol. Eng. 2019; 13: 78. https://doi.org/10.1186/s13036-019-0206-z
  18. Belzhelarskaya C.N. Baculovirus exression systems for recombinant protein production in insect and mammalian cells. Molekulyarnaya biologiya. 2011; 45(1): 142–59. EDN: https://www.elibrary.ru/ndjchz (in Russian)
  19. Changotra H., Vij A. Rotavirus virus-like particles (RV-VLPs) vaccines: An update. Rev. Med. Virol. 2017; 27(6). https://doi.org/10.1002/rmv.1954
  20. Shoja Z., Jalilvand S., Latifi T., Roohvand F. Rotavirus VP6: involvement in immunogenicity, adjuvant activity, and use as a vector for heterologous peptides, drug delivery, and production of nano-biomaterials. Arch. Virol. 2022; 167(4): 1013–23. https://doi.org/10.1007/s00705-022-05407-9
  21. Heinimäki S., Tamminen K., Hytönen V.P., Malm M., Blazevic V. Rotavirus inner capsid VP6 acts as an adjuvant in formulations with particulate antigens only. Vaccines (Basel). 2020; 8(3): 365. https://doi.org/10.3390/vaccines8030365
  22. Macpherson A.J., Mccoy K.D., Johansen F.E., Brandtzaeg P. The immune geography of IgA induction and function. Mucosal. Immunol. 2008; 1(1): 11–22. https://doi.org/10.1038/mi.2007.6
  23. Mantis N.J., Rol N., Corthesy B. Secretory IgA’s complex roles in immunity and mucosal homeostasis in the gut. Mucosal. Immunol. 2011; 4(6): 603–11. https://doi.org/10.1038/mi.2011.41

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. The result of transfection of the Sf-9 cell line with bacmid DNA containing the genes of RVA proteins VP6 and VP2-eGFP: а – not infected Sf-9 cell line in transmitted light; b, c – Sf-9 cell line infected with baculovirus first (b) and third (c) passages in fluorescence field.

Download (647KB)
Indexing metadata
3. Fig. 2. Sf-21 cells infected with 10–4 virus dilution: а – in transmitted light; b, c, d – plaques in fluorescence field.

Download (1MB)
Indexing metadata
4. Fig. 3. Electron micrograph of RVA VP2/VP6 VLPs (a) and RVA (b). ×25,000 magnification.

Download (482KB)
Indexing metadata
5. Fig. 4. Protein analysis of RVA VP2/VP6 VLP by SDS-PAAGE. Two bands are visible in 12% gel that correspond to molecular weight of the RVA structural proteins VP2 and VP6 (120 and 45 kDa). No additional protein bands are present.

Download (180KB)
Indexing metadata
6. Fig. 5. Comparative analysis of specific recognition of rec-VP6 (blue columns) and RVA VP2/VP6 VLP (red columns) which were utilized in ELISA as antigen for the detection of IgG (а), IgM (b) и IgA (c) antibodies in mouse serum samples.

Download (39KB)
Indexing metadata
7. Fig. 6. Distribution of IgG (a) and IgA (b) antibodies to RVA in the serum samples of newborn dwarf piglets. RVA VP2/VP6 VLPs were used as antigen in ELISA, serum samples were collected before and after of each vaccination. The geometric mean values for reverse antibody titers are shown.

Download (56KB)
Indexing metadata
8. Fig. 7. Titration curves of guinea pigs serum samples obtained in VLP-based ELISA representing the levels of RVA-specific IgG antibodies. RVA VP2/VP6 VLPs were used as antigen in ELISA, serum samples were collected after first (a), second (b) and third (c) vaccination, respectively. Sera from non-vaccined animals were used as negative controls.

Download (146KB)
Indexing metadata

Copyright (c) 2023 Filatov I.E., Tsibezov V.V., Balandina M.V., Norkina S.N., Latyshev O.E., Eliseeva O.V., Cherepushkin S.A., Verkhovsky O.A., Grebennikova T.V.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies