Expression of the Hepatitis C Virus core-NS3 Fusion Protein on the Surface of  Bacterial Ghosts: Prospects for Vaccine Production

Minoosadat  Tayebinia; Sedigheh  Sharifzadeh; Gholamreza  Rafiei Dehbidi; Farahnaz  Zare; Reza  Ranjbaran; Amir  Rahimi; Mohammad Reza  Miri; Mehdi  Mirzakhani; Abbas  Behzad-Behbahania

doi:10.18502/ajmb.v15i3.12927

Minoosadat Tayebinia Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Sedigheh Sharifzadeh Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Gholamreza Rafiei Dehbidi Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Farahnaz Zare Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Reza Ranjbaran Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Amir Rahimi Bioinfirmatic and Computational Biology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Mohammad Reza Miri The Persian Gulf Marine Biotechnology Research Center, the Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
Mehdi Mirzakhani Fars Blood Transfusion Organization, Shiraz, Iran
Abbas Behzad-Behbahania Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

DOI: https://doi.org/10.18502/ajmb.v15i3.12927

Keywords: Antigen presentation, Epitopes, Flow cytometry, Hepatitis C, Plasmids

Abstract

Background: Antigen presentation using bacterial surface display systems, on one hand, has the benefits of bacterial carriers, including low-cost production and ease of manipulation. On the other hand, the bacteria can help in stimulating the immune system as an adjuvant. For example, using bacterial surface display technology, we developed a hepatitis C virus (HCV) multiple antigens displaying bacteria's surface and then turned it into a bacterial ghost.

Methods: The HCV core and NS3 proteins' conserved epitopes were cloned into the AIDA gene plasmid as an auto transporter. The recombinant plasmid was then transformed into Escherichia coli (E. coli) Bl21 (DE3). Recombinant bacteria were then turned into a bacterial ghost, an empty cell envelope. Whole-cell ELISA, flow cytometry, and Western blot techniques were used for monitoring the expression of proteins on the surface of bacteria.

Results: A fusion protein of HCV core-NS3-AIDA was successfully expressed on the E. coli Bl21 (DE3) surface and confirmed by western blotting, Enzyme-Linked Immunosorbent Assay (ELISA), and flow cytometry detection techniques.

Conclusion: The presence of HCV antigens on non-pathogen bacteria surfaces holds promise for developing safe and cost-benefit-accessible vaccines with optimal intrinsic adjuvant effects and exposure of heterologous antigens to the immune system.