Abstract
Recently, it has been shown that HCV core proteins (HCcAg) with C-terminal deletions assemble in vitro into virus-like particles (VLPs) in the presence of structured RNA molecules. Results presented in this work showed that a truncated HCcAg variant covering the first 120 aa (HCcAg.120) with a 32 aa N-terminal fusion peptide (6xHistag-XpressTMepitope) interacts with plasmid DNA vaccine. Interestingly, the buoyant density of VLPs containing HCcAg.120 in CsCl gradients changed from 1.15-1,17 g mLˉ1 to 1.30-1.34 g mLˉ1 after addition of plasmid DNA to assembly reactions. In addition, a delay in electrophoretic mobility of HCcAg.120-plasmid samples on agarose gels was observed indicating a direct interaction between VLPs and nucleic acids. Remarkably, addition of either plasmid DNA or tRNA to assembly reactions leaded to heterogeneous and larger VLPs formation than those observed in HCcAg.120 assembly reactions. VLPs containing HCcAg.120 induced a specific IgG antibodies in mice that reacted with hepatocytes from HCV-infected patients. VLPs obtained in this work would be important to elucidate the mechanisms behind the ability of HCcAg to assemble into a nucleocapsid structure. Besides, the capacity of particles containing HCcAg.120 to interact with nucleic acids could be used in the development of DNA vaccines and viral vectors based on these particles.
Highlights
Hepatitis C virus (HCV) is a major worldwide health problem
A previous study had demonstrated that a DNA vaccine containing the structural region of HCV induces a strong humoral and cellular immune response in various animal models[12,13]
Since particulate antigens have been proposed as potential carriers for DNA vaccines[14], the capacity of HCcAg.120 to interact with pIDKE2 plasmid was investigated
Summary
HCV is a member of the Hepacivirus genus of the Flaviviridae family[1]. The genome is a single-stranded positive-sense RNA of approximately 9600 nt and encodes a polyprotein with a single open reading frame (ORF) of 3008–3033 aa which is processed by host and viral proteases[2]. The structural proteins (core, E1, E2 and potentially p7) precede the nonstructural proteins (NS2, 3, 4A, 4B, 5A and 5B) in the polyprotein. The biogenesis of HCV core protein (HCcAg) is dependent on the interaction of the signal sequence of nascent polypeptide with the endoplasmic reticulum (ER) membrane[3]. It has been suggested that HCcAg function to encapsidate the viral genome within a nucleocapsid particle, little is known about the in vivo assembly pathway or structure of the HCV nucleocapsid and virion[4]
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