Abstract
The envelope protein (E1–E2) of Hepatitis C virus (HCV) is a major component of the viral structure. The glycosylated envelope protein is considered to be important for initiation of infection by binding to cellular receptor(s) and also known as one of the major antigenic targets to host immune response. The present study was aimed at identifying mouse monoclonal antibodies which inhibit binding of virus like particles of HCV to target cells. The first step in this direction was to generate recombinant HCV-like particles (HCV-LPs) specific for genotypes 3a of HCV (prevalent in India) using the genes encoding core, E1 and E2 envelop proteins in a baculovirus expression system. The purified HCV-LPs were characterized by ELISA and electron microscopy and were used to generate monoclonal antibodies (mAbs) in mice. Two monoclonal antibodies (E8G9 and H1H10) specific for the E2 region of envelope protein of HCV genotype 3a, were found to reduce the virus binding to Huh7 cells. However, the mAbs generated against HCV genotype 1b (D2H3, G2C7, E1B11) were not so effective. More importantly, mAb E8G9 showed significant inhibition of the virus entry in HCV JFH1 cell culture system. Finally, the epitopic regions on E2 protein which bind to the mAbs have also been identified. Results suggest a new therapeutic strategy and provide the proof of concept that mAb against HCV-LP could be effective in preventing virus entry into liver cells to block HCV replication.
Highlights
Hepatitis C virus (HCV) is the major etiological agent of non-A, non-B hepatitis that infects almost 200 million people worldwide [1]
70–80% of HCV patients develop chronic hepatitis of which 20–30% leads to liver disease, cirrhosis and hepatocellular carcinoma [2]
The structural proteins of HCV are comprised of the core protein (,21 kDa) and two envelope glycoproteins E1 (,31 kDa) and E2 (,70 kDa) [3,4,5]
Summary
Hepatitis C virus (HCV) is the major etiological agent of non-A, non-B hepatitis that infects almost 200 million people worldwide [1]. HCV is a major cause of post transfusion and communityacquired hepatitis. 70–80% of HCV patients develop chronic hepatitis of which 20–30% leads to liver disease, cirrhosis and hepatocellular carcinoma [2]. The virion contains a positive-sense single stranded RNA genome of approximately 9.6 kb that consists of a highly conserved 59 non coding region followed by a long open reading frame of 9,030 to 9,099 nucleotides (nts). It is translated into a single polyprotein of 3,010 to 3030 amino acids [3,4]. E1 and E2 undergo post translational modifications by extensive N-linked glycosylation and are responsible for cell binding and entry [6,7,8,9,10,11,12,13,14,15]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
