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JVI Accepts, published online ahead of print on 12 December 2007
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J. Virol. doi:10.1128/JVI.02093-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

HEPATITIS C VIRUS GENOTYPE 1a GROWTH AND INDUCTION OF AUTOPHAGY

Malika Ait-Goughoulte, Tatsuo Kanda, Keith Meyer, Jan S. Ryerse, Ratna B. Ray, and Ranjit Ray*

Departments of Internal Medicine, Pathology, and Molecular Microbiology & Immunology, Saint Louis University, Missouri

* To whom correspondence should be addressed. Email: rayr{at}slu.edu.


  Abstract

We have previously reported that immortalized human hepatocytes (IHH) support the generation of infectious hepatitis C virus (HCV) genotype 1a (clone H77). In the present study, we have investigated the growth of HCV genotype 1a (clone H77) through serial passage, and accompanying changes in IHH in response to infection. Eleven serial passage of HCV genotype 1a (clone H77) in IHH have been completed. Virus replication was ascertained from the presence of HCV specific sequences, detection of core antigen, virus genome copy number, and virus titer in IHH culture fluid. Electron microscopy suggested that HCV infection induces autophagic vacuole formation in IHH. Fluorescence microscopy displayed localization of autophagic markers, microtubule associated protein-1 light chain-3 (LC3) and Apg5, on the vacuoles of HCV infected hepatocytes. Taken together, our results suggested that HCV genotype 1a (clone H77) can be serially passaged in IHH, and HCV infection induces an autophagic response in hepatocytes.




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