A reporter cell line for rapid and sensitive evaluation of hepatitis C virus infectivity and replication

Iro, M., Witteveldt, J., Angus, A.G.N., Woerz, I., Kaul, A., Bartenschlager, R. and Patel, A.H. (2009) A reporter cell line for rapid and sensitive evaluation of hepatitis C virus infectivity and replication. Antiviral Research, 83(2), pp. 148-155. (doi:10.1016/j.antiviral.2009.04.007) (PMID:19397930)

Full text not currently available from Enlighten.

Abstract

The human pathogen hepatitis C virus (HCV) is associated with chronic liver disease. The recent development of the cell culture infectious HCV (HCVcc) system has opened up avenues for detailed studies on the life cycle of the virus and its interaction with the host cell. Current methods to quantitate virus infectivity in cell culture are time-consuming and labor-intensive. This study describes the generation of a cell-based secreted alkaline phosphatase (SEAP) reporter assay to facilitate in vitro studies of HCV infection and replication. This assay is based on a novel reporter cell line stably expressing the enhanced green fluorescent protein (EGFP) fused in-frame to the secreted alkaline phosphatase (SEAP) via a recognition sequence of the viral NS3/4A serine protease. The SEAP reporter from a similar construct has previously been shown to be released from the fusion protein and be secreted into the extracellular culture medium following cleavage by the viral NS3/4A protease. The reporter cell line enabled rapid and sensitive quantification of HCV infection and viral replication in cell culture. The utility of this system for investigating virus entry, and for high throughput screening of entry inhibitors and other antiviral compounds was demonstrated using several inter- and intra-genotypic chimeras of HCV.

Item Type:Articles
Keywords:Hepatitis C virus; HCV; chimera; reporter cell line; SEAP; NS3/4A protease
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Angus, Dr Allan and Witteveldt, Dr Jeroen and Patel, Professor Arvind
Authors: Iro, M., Witteveldt, J., Angus, A.G.N., Woerz, I., Kaul, A., Bartenschlager, R., and Patel, A.H.
Subjects:Q Science > QR Microbiology > QR355 Virology
College/School:College of Medical Veterinary and Life Sciences
Journal Name:Antiviral Research
Publisher:Elsevier
ISSN:0166-3542
ISSN (Online):1872-9096
Published Online:04 May 2009

University Staff: Request a correction | Enlighten Editors: Update this record