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Characterization of Coxsackievirus B4 virus-like particles VLP produced by the recombinant baculovirus-insect cell system expressing the major capsid protein

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Abstract

Coxsackievirus B4 (CV-B4) is suspected to be an environmental factor that has the intrinsic capacity to damage the pancreatic beta cells and therefore causes insulitis and type 1 diabetes (T1D). Although vaccination against CV-B4 could reduce the incidence of this chronic auto-immune disease, there is currently no therapeutic reagent or vaccine in clinical use. By the employment of the Bac-to-Bac® vector system to express the major viral capsid protein, we contributed towards the development of a CV-B4 vaccine by producing CV-B4 virus-like particles (VLPs) from recombinant baculovirus in infected insect cells. In fact Western blot and Immunofluorescence analysis detected the viral protein 1 (VP1) in the cells resulting from the construction of a recombinant bacmid DNA carrying the key immunogenic protein then transfected in the insect cells. Sucrose gradient ultracentrifugation fractions of the infected cell lysates contained the recombinant protein and the electron microscopy demonstrated the presence of VLPs in these sucrose fractions. This study clearly shows for the first time the expression of CVB4 VP1 structure protein alone can form VLPs in the baculovirus-infected insect cell keeping conserved both characteristics and morphology.

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Acknowledgements

Authors are thankful for the skillful technical assistance provided by Antonio Varas, Dolores Rodríguez, Ana Oña and Cristina Patiño from the CNB Confocal and Electron Microscopy services. Authors acknowledge the Tunisian Ministry of Higher Education and Scientific Research (UR17ES30) and the Deanship of Scientific Research at King Faisal University for the financial support under Nasher Track (Grant No. 186390).

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Authors and Affiliations

  1. Unité de Recherche UR17ES30 “Génomique, Biotechnologie et Stratégies Antivirales”, Institut Supérieur de Biotechnologie, Université de Monastir, BP74, Avenue Tahar Hadded, 5000, Monastir, Tunisia

    Ikbel Hadj Hassine, Jawhar Gharbi, Bechr Hamrita & Manel Ben M’hadheb

  2. Department of Molecular and Cellular Biology, Centro Nacional de Biotecnología-CSIC, Cantoblanco, 28049, Madrid, Spain

    Ikbel Hadj Hassine & José Francisco Rodríguez

  3. Department of Biological Sciences, College of Science, King Faisal University, P.O. Box 380, Al-Ahsa, 31982, Saudi Arabia

    Jawhar Gharbi & Mohammed A. Almalki

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  1. Ikbel Hadj Hassine
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Correspondence to Jawhar Gharbi.

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Hassine, I.H., Gharbi, J., Hamrita, B. et al. Characterization of Coxsackievirus B4 virus-like particles VLP produced by the recombinant baculovirus-insect cell system expressing the major capsid protein. Mol Biol Rep 47, 2835–2843 (2020). https://doi.org/10.1007/s11033-020-05333-6

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