Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon

Kato, Takanobu; Date, Tomoko; Miyamoto, Michiko; Furusaka, Akihiro; Tokushige, Katsutoshi; Mizokami, Masashi; Wakita, Takaji

doi:10.1053/j.gastro.2003.09.023

Cited by 537 publications

(498 citation statements)

References 24 publications

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“…In contrast to the subgenomic replicons, the JFH-1 HCV consensus genome is capable of high-level replication without requiring adaptive mutations. 27 More importantly, JFH-1 genomes were shown to produce HCV viral particles (HCVcc) capable of infection in cell culture and animals (chimpanzee and mice with humanized livers). Infectivity of the JFH-1 replicon was significantly increased by creating virus chimeras by replacing the core to NS2 region with that of the J6 genotype 2a isolate.…”

Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

“…14,25 A different situation emerged when the first genotype 2a consensus genome was established. 26,27 A subgenomic replicon constructed from a clone called JFH-1, isolated from a Japanese patient with fulminant hepatitis C, replicated up to 20-fold higher in Huh7 cells as compared to Con1 replicons, and did not require adaptive mutations for efficient replication in vitro. 27 Transfection of Huh7 and Huh7.5.1 cells with the in vitro-transcribed fulllength JFH-1 genome or a recombinant chimeric genome with another genotype 2a isolate, J6, resulted in the secretion of viral particles that were infectious in cultured cells, in chimeric mice, and in chimpanzees.…”

Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

“…26,27 A subgenomic replicon constructed from a clone called JFH-1, isolated from a Japanese patient with fulminant hepatitis C, replicated up to 20-fold higher in Huh7 cells as compared to Con1 replicons, and did not require adaptive mutations for efficient replication in vitro. 27 Transfection of Huh7 and Huh7.5.1 cells with the in vitro-transcribed fulllength JFH-1 genome or a recombinant chimeric genome with another genotype 2a isolate, J6, resulted in the secretion of viral particles that were infectious in cultured cells, in chimeric mice, and in chimpanzees. [28][29][30][31] The infectivity of cells could be neutralized with antibodies against the HCV entry receptor CD81, antibodies against E2, or immunoglobulins from chronically infected patients.…”

Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

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Experimental models for hepatitis C viral infection

Boonstra¹,

Laan²,

Vanwolleghem³

et al. 2009

Hepatology

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Hepatitis C virus (HCV) infection is a leading cause of chronic liver disease. The majority of infected individuals develop a persistent infection, which is associated with a high risk of liver cirrhosis and hepatocellular carcinoma. Since its discovery 20 years ago, progress in our understanding of this virus has been suboptimal due to the lack of good model systems. However, in the past decade this has greatly accelerated with the development of various in vitro cell culture systems and in vivo small-animal models. These systems have made a major impact on the field of HCV research, and have provided important breakthroughs in our understanding of HCV infection and replication. Importantly, the in vitro cell culture systems and the small-animal models have allowed preclinical testing of numerous novel antiviral compounds for the treatment of chronic HCV infection. In this article, we give an overview of current models, discuss their limitations, and provide future perspectives for research directed at the prevention and cure of hepatitis C. (HEPATOLOGY 2009;50:1646-1655.) Hepatitis C Virus Pathology and BiologyThe hepatitis C virus (HCV) is considered a successful pathogen in establishing persistent infections by evading the immune system. Only a fraction of acutely infected individuals are able to clear the infection spontaneously, whereas about 80% of the infected individuals develop a chronic infection. 1,2 The symptoms are initially mild in these persistently infected patients, and it may take decades before the serious consequences of chronic HCV infection become apparent. Patients with chronic HCV are at increased risk for developing liver fibrosis, cirrhosis, and/or hepatocellular carcinoma. Currently, these longterm complications of chronic HCV infection are the leading indication for liver transplantation. 3,4 Because of the high incidence of new infections by blood transfusions in the 1980s before discovery of the virus, and because morbidity associated with chronic HCV infections generally takes decades to develop, it is expected that the burden of disease in the near future will rise dramatically.HCV is an enveloped flavivirus, with a positivestranded RNA genome of approximately 9600 nucleotides and is composed of a single open reading frame, which encodes a polyprotein precursor of approximately 3000 amino acids. The coding region is flanked by 5Ј and 3Ј noncoding regions, which are important for the regulation of genomic duplication as well as initiation of translation. The single polyprotein is cleaved by host and viral proteases into individual structural and nonstructural (NS) proteins (Fig. 1A). Replication of the HCV genome involves the synthesis of a full-length negative-stranded RNA intermediate, which in turn provides a template for the de novo production of positive-stranded RNA. Both these synthesis steps are mediated by the viral RNA-dependent RNA polymerase NS5B. 5-7 NS5B lacks proofreading abilities, and this leads to a high mutation rate and the generation of numerous quasispecies. ...

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Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

Section: Cell-based In Vitro Hcv Systemsmentioning

confidence: 99%

See 1 more Smart Citation

Experimental models for hepatitis C viral infection

Boonstra¹,

Laan²,

Vanwolleghem³

et al. 2009

Hepatology

View full text Add to dashboard Cite

show abstract

“…Moreover, the replicon system has been exploited for analyses of the effect of cytokines on HCV RNA replication 46,47 and was instrumental in a recent series of elegant studies looking at host proteins involved in HCV RNA replication as well as mechanisms of evasion from innate immune responses. [48][49][50][51][52][53] Since the original reports of functional genotype 1b replicons, replicons for genotype 1a 54 and 2a 55 as well as derivatives expressing easily quantifiable marker enzymes in a separate cistron have been made to facilitate genetic studies as well as drug screening and evaluation. [56][57][58] In addition, full-length replicons and HCV genomes efficiently replicating in tissue culture have been developed, 44,59,60 and the spectrum of permissive host cells has been expanded.…”

Section: The Replicon Systemmentioning

confidence: 99%

Hepatitis C virus comes full circle: Production of recombinant infectious virus in tissue culture

Berke

Moradpour

2005

Hepatology

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“…Continued search on HCV isolates with high in vitro replication efficiency yielded the most famous HCV cDNA clone, JFH-1 (genotype 2a). JFH-1 was isolated from a fulminant hepatitis patient by Takaji Wakita (Tokyo Metropolitan Institute of Neuroscience) in 2001 (Kato et al, 2001), exhibiting high efficient replication capability without additional adaptive mutations (Kato et al, 2003). In 2005, studies from Rice and Wakita were simultaneously published to demonstrate the recombinant HCV virus derived from JFH-1 or chimeric genomes based on JFH-1 backbone were infectious in Huh7 cells (Lindenbach et al, 2005;Wakita et al, 2005).…”

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confidence: 99%