Hepatitis B and C virus coinfection: A novel model system reveals the absence of direct viral interference

Bellecave, Pantxika; Gouttenoire, Jérôme; Gajer, M.; Brass, Volker; Koutsoudakis, George; Blum, Hubert E.; Bartenschlager, Ralf; Nassal, Michael; Moradpour, Darius

doi:10.1002/hep.22951

Cited by 142 publications

(135 citation statements)

References 36 publications

(57 reference statements)

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“…S9C) replication. These data indicate that HBV and HCV replicate in the same cells without evidence of direct interference in vitro (27). Superinfection exclusion generally is restricted to homologous viruses (28), whereas nonrelated viruses are able to replicate normally.…”

Section: Resultsmentioning

confidence: 76%

Complete replication of hepatitis B virus and hepatitis C virus in a newly developed hepatoma cell line

Yang

Zuo

Wang

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The absence of a robust cell culture system for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection has limited the analysis of the virus lifecycle and drug discovery. We have established a hepatoma cell line, HLCZ01, the first cell line, to the authors' knowledge, supporting the entire lifecycle of both HBV and HCV. HBV surface antigen (HBsAg)-positive particles can be observed in the supernatant and the lumen of the endoplasmic reticulum of the cells via electron microscopy. Interestingly, HBV and HCV clinical isolates propagate in HLCZ01 cells. Both viruses replicate in the cells without evidence of overt interference. HBV and HCV entry are blocked by antibodies against HBsAg and human CD81, respectively, and the replication of HBV and HCV is inhibited by antivirals. HLCZ01 cells mount an innate immune response to virus infection. The cell line provides a powerful tool for exploring the mechanisms of virus entry and replication and the interaction between host and virus, facilitating the development of novel antiviral agents and vaccines.cell culture model | primary human hepatocytes | cccDNA | interferon | ISGs M ore than 500 million people worldwide are persistently infected with hepatitis B virus (HBV) and/or hepatitis B virus (HCV) and are at risk of developing chronic liver diseases (1). There is no vaccine against HCV, and many patients who are persistently infected by HBV or HCV do not respond to currently available therapies (2, 3). Improved understanding of the biology and pathogenesis of these infections is required for the development of vaccine and antiviral drugs (4). The inability to grow HBV and HCV efficiently in cell culture has presented a major obstacle to understanding the virus lifecycle and pathogenesis and to developing improved therapeutics.HBV is a member of the hepadnavirus families, and its genome is a relaxed circular, partially double-stranded DNA molecule. The negative strand has an invariable length of ∼3.2 kb, and the positive strand is 50-100% of this length. Several key issues about the biology of HBV remain to be explored, including the identification of the cellular receptors, the role of the X gene, and the mechanisms by which the viral minichromosome is formed. Covalently closed circular DNA (cccDNA) is responsible for the establishment of viral infection and persistence. Understanding the mechanisms underlying cccDNA formation and regulation is critical for understanding the HBV pathogenesis and finding a cure for hepatitis B. HepG2.2.15 cells derived from the hepatoma cell line HepG2 transfected with the full genome of HBV have been used to study HBV replication (5). Primary human hepatocytes (PHH) are susceptible to HBV infection (6, 7), but the use of this model is hampered by the limited availability and unpredictable variability of human liver. Several human hepatoma cell lines support HBV replication after HBV DNA transfection, and overexpression of sodium-taurocholate cotransporting polypeptide (NTCP) in HepG2 and Huh7 cells can render these cells able t...

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Section: Resultsmentioning

confidence: 76%

Complete replication of hepatitis B virus and hepatitis C virus in a newly developed hepatoma cell line

Yang

Zuo

Wang

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…RNA replication assay using a JFH1 subgenomic replicon harboring the firefly luciferase was performed as described. 18,21 Jc1 cell culture-derived HCV (HCVcc) was produced as described. 19 The 50% tissue culture infective dose (TCID 50 ) was determined as described.…”

Section: Methodsmentioning

confidence: 99%

Quantitative proteomics identifies the membrane-associated peroxidase GPx8 as a cellular substrate of the hepatitis C virus NS3-4A protease

et al. 2013

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The hepatitis C virus (HCV) NS3-4A protease is not only an essential component of the viral replication complex and a prime target for antiviral intervention but also a key player in the persistence and pathogenesis of HCV. It cleaves and thereby inactivates two crucial adaptor proteins in viral RNA sensing and innate immunity, mitochondrial antiviral signaling protein (MAVS) and TRIF, a phosphatase involved in growth factor signaling, T-cell protein tyrosine phosphatase (TC-PTP), and the E3 ubiquitin ligase component UV-damaged DNA-binding protein 1 (DDB1). Here we explored quantitative proteomics to identify novel cellular substrates of the NS3-4A protease. Cell lines inducibly expressing the NS3-4A protease were analyzed by stable isotopic labeling using amino acids in cell culture (SILAC) coupled with protein separation and mass spectrometry. This approach identified the membrane-associated peroxidase GPx8 as a bona fide cellular substrate of the HCV NS3-4A protease. Cleavage by NS3-4A occurs at Cys 11, removing the cytosolic tip of GPx8, and was observed in different experimental systems as well as in liver biopsies from patients with chronic HCV. Overexpression and RNA silencing studies revealed that GPx8 is involved in viral particle production but not in HCV entry or RNA replication. Conclusion: We provide proof-of-concept for the use of quantitative proteomics to identify cellular substrates of a viral protease and describe GPx8 as a novel proviral host factor targeted by the HCV NS3-4A protease. (HEPATOLOGY 2014;59:423-433)

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“…Using a novel in vitro system, a recent study well demonstrated the interaction between HBV and HCV [42]. In this model, stable Huh-7 cell lines inducibly replicating HBV were transfected with selectable HCV replicons or infected with cell culture-derived HCV.…”

Section: Viral Interactions Between Hbv and Hcv And Their Impact On Tmentioning

confidence: 99%

Dual chronic hepatitis B virus and hepatitis C virus infection

Liu

Chen

2009

Hepatol Int

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Dual hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are common in HBV or HCV endemic areas. However, several clinical and pathogenetic issues remain unresolved. First, clinical and in vitro studies suggest the interactions between two viruses. The dynamics of the interaction in untreated setting versus treated setting and its influence on the long-term outcomes await further studies. A key issue regarding viral interactions is whether modulation of infection occurs in the same dually infected individual hepatocyte of the liver. Clarifying this issue may help to understand the reciprocal interference between HCV and HBV and provide clues for future immunopathogenetic studies. Second, the prevalence and clinical significance of coexisting occult HBV infection in patients with chronic HCV infection need further investigations. Third, combination therapy of peginterferon alfa2a and ribavirin appears to be just as effective and safe for the treatment of hepatitis B surface antigen (HBsAg)-positive patients chronically infected with active chronic hepatitis C as it is in patients with HCV monoinfection. Nevertheless, one-third of dually infected patients with nondetectable serum HBV DNA-level pretreatment developed HBV reactivation posttreatment. How to prevent and treat this reactivation should be clarified. Furthermore, about 10% of the dually infected patients lost HBsAg. Underlying mechanisms await further investigations. Finally, the optimal treatment strategies for dually infected patients with hepatitis B e antigen-positive chronic hepatitis B should be identified in future clinical trials.

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Hepatitis B and C virus coinfection: A novel model system reveals the absence of direct viral interference

Cited by 142 publications

References 36 publications

Complete replication of hepatitis B virus and hepatitis C virus in a newly developed hepatoma cell line

Complete replication of hepatitis B virus and hepatitis C virus in a newly developed hepatoma cell line

Quantitative proteomics identifies the membrane-associated peroxidase GPx8 as a cellular substrate of the hepatitis C virus NS3-4A protease

Dual chronic hepatitis B virus and hepatitis C virus infection

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