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

• Published in: Molecular biology reports Vol. 47; no. 4; pp. 2835 - 2843
• Main Authors: Hassine, Ikbel Hadj, Gharbi, Jawhar, Hamrita, Bechr, Almalki, Mohammed A., Rodríguez, José Francisco, Ben M’hadheb, Manel
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.04.2020; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Animals; Autoimmune diseases; Baculoviridae - genetics; Beta cells; Biomedical and Life Sciences; Capsid protein; Capsid Proteins - genetics; Capsid Proteins - metabolism; Cell Line; Coxsackievirus; Coxsackieviruses; Cytology; Diabetes mellitus (insulin dependent); Electron microscopy; Enterovirus B, Human - genetics; Enterovirus B, Human - metabolism; Enterovirus B, Human - pathogenicity; Environmental factors; Histology; Humans; Immunofluorescence; Immunogenicity; Immunological diseases; Insect cells; Insecta; Insulitis; Life Sciences; Lysates; Morphology; Original Article; Physical characteristics; Protein Engineering - methods; Protein structure; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sucrose; Ultracentrifugation; Vaccination; Vaccines; Virion - genetics; Virus-like particles; VP1 protein; Baculovirus expression system; Recombinant VP1; Virus-like particles; Coxsackievirus B4
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-020-05333-6

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.