Abstract
In 2005, the first robust hepatitis C virus (HCV) infectious cell culture system was developed based on the HCV genotype 2a JFH-1 molecular clone and the human-derived hepatoma cell line Huh7. Although much effort has been made to dissect and expand the repertoire of JFH-1-derived clones, less attention has been given to the host cell despite the intriguing facts that thus far only Huh7 cells have been found to be highly permissive for HCV infection and furthermore only a limited number of Huh7 cell lines/stocks appear to be fully permissive. As such, we compiled a panel of Huh7 lines from disparate sources and evaluated their permissiveness for HCV infection. We found that although Huh7 lines from different laboratories do vary in morphology and cell growth, the majority (8 out of 9) were highly permissive for infection, as demonstrated by robust HCV RNA and de novo infectious virion production following infection. While HCV RNA levels achieved in the 8 permissive cell lines were relatively equivalent, three Huh7 lines demonstrated higher infectious virion production suggesting these cell lines more efficiently support post-replication event(s) in the viral life cycle. Consistent with previous studies, the single Huh7 line found to be relatively resistant to infection demonstrated a block in HCV entry. These studies not only suggest that the majority of Huh7 cell lines in different laboratories are in fact highly permissive for HCV infection, but also identify phenotypically distinct Huh7 lines, which may facilitate studies investigating the cellular determinants of HCV infection.
Highlights
Worldwide,170 million individuals are infected with hepatitis C virus (HCV), an enveloped positive-strand RNA virus that infects and replicates in the hepatocytes of the liver
Morphological examination and growth kinetics Since being isolated in 1982 from a Japanese male with a welldifferentiated hepatocellular carcinoma [16], Huh7 cells have become widely used for the study of liver-related diseases and were fundamental to the development of the HCV infectious cell culture system [4,5,6]
We evaluated eight Huh7 lines obtained from disparate sources including the Huh7/scr cells, as these cells have been shown to be highly permissive for HCVcc infection [8,9]
Summary
Worldwide, ,170 million individuals are infected with hepatitis C virus (HCV), an enveloped positive-strand RNA virus that infects and replicates in the hepatocytes of the liver. In 2003 Wakita and colleagues cloned a HCV consensus genotype 2a genome (JFH-1) from a Japanese Fulminant Hepatitis patient [3], which was subsequently found to produce infectious HCV in Huh cell cultures [4,5,6]. This breakthrough was the result of two culminating factors: (1) identification of a HCV clone capable of replicating and assembling infectious particles in cell culture and (2) discovery that Huh cells are permissive for HCV infection. We assembled a panel of Huh cell lines from disparate sources and assessed their permissiveness for HCV infection
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