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A Cell Culture Model for Persistent HCV Infection

  • Victoria Castro
  • Ginés Ávila-Pérez
  • Lidia Mingorance
  • Pablo GastaminzaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1911)

Abstract

Chronic hepatitis C virus (HCV) infection affects millions of humans throughout the globe, causing liver disease and hepatocellular carcinoma. Persistence of the virus in the infected host can last for decades as a result of a faulty immune response that fails to clear the virus while constituting a major player in viral pathogenesis. In addition to evading immune responses, HCV has evolved intracellular survival strategies that enable persistent replication without directly killing the host cell.

After the generation of cell culture infection models for HCV, the knowledge about this virus and host-virus interactions acquired in the last decade has been greatly increased. Interestingly, persistent infection can also be established in cell culture. This model recapitulates persistent HCV RNA replication and viral protein expression as well as infectious progeny virus assembly and secretion and may be used to study not only these aspects of the virus replication cycle but also to study host-virus interactions in a model of prolonged HCV infection. In this chapter, we describe a methodology to generate persistently HCV-infected cultures and to monitor viral load and progeny virus production. Also, we provide generic protocols to study the impact of chemical compounds and host-targeting shRNAs to illustrate the applications of this model in the study of HCV infection in cell culture.

Key words

Hepatitis C Persistent infection Cell culture Antiviral Host-virus interaction 

Notes

Acknowledgment

This protocol is based on work initially developed at Dr. Francis V. Chisari’s laboratory at The Scripps Research Institute (La Jolla, CA) in collaboration with Dr. Takaji Wakita’s group at the Tokyo Metropolitan Institute of Medical Science (Tokyo, Japan) and was originally generated and improved with the essential contribution of Jin Zhong (Pasteur Institute in Shanghai), Sharookh Kapadia (Genentech), Guofeng Cheng (Gilead), Susan Uprichard (University of Illinois, Chicago), and specially Stefan Wieland (University Hospital of Basel). We would like to thank Gema Calvo and Martina Friesland (CNB-CSIC) for excellent technical assistance and Dr. Urtzi Garaigorta for critically reviewing the manuscript. This work was supported by grant SAF2014-52400-R from the Spanish Ministry of Economy.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Victoria Castro
    • 1
  • Ginés Ávila-Pérez
    • 1
  • Lidia Mingorance
    • 1
  • Pablo Gastaminza
    • 1
    Email author
  1. 1.Department of Molecular and Cellular BiologyCentro Nacional De Biotecnología-Consejo Superior de Investigaciones Científicas (CNB-CSIC)MadridSpain

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