Production of rabbit specific antibodies against conserved and potentially immunogenic peptides of The HCV envelope glycoprotein E2: Novel candidate peptide vaccine against HCV infection

Hepatitis C Virus (HCV) is a worldwide public health problem that affects more than 70% of the estimated 170 million people with chronic hepatitis. It induces chronic hepatic lesions that lead to severe fibrosis and cirrhosis, hepatic failure, or hepatocellular carcinoma. Envelope (E1 /E2) proteins of HCV may generate neutralizing antibodies. This study aims to evaluate and improve the bifunctional chimeric vaccine expression system based on the HBsAg protein as a site for the introduction of heterologous proteins (chimeric HBsAg). Therefore, it was designed to assess the immunogenic properties of conserved peptides derived from E2 HVR1 regions. Transfection assays were carried out in 25cm2 bottles and 6-well plates containing HuH7 cells (Hepatocellular carcinoma cells) and HEK-293T (Human Embryonic Kidney 293 cells associated with the SV-40 T antigen) in medium with low glucose concentration and high glucose concentration respectively, following specifications of the American Collection for the type of culture. The supernatants and cell lysates were collected and used for evaluation for the presence of HBsAg by immunoenzymatic assay and Western Blot. Samples obtained from transfections were subjected to denaturing polyacrylamide gel electrophoresis. Only two samples of patients reacted with the peptide 412-419 BSA in different dilutions. All peptides are shown to be highly immunogenic and neutralizing with detectable levels of specific antibodies.