Hepatitis C virus infection of human hepatoma cell line 7721 in vitro

Song, Zhiqiang; Fei, Hao; Feng, Ming; Ma, Qiaoyu; Liu, Guodong

doi:10.3748/wjg.7.5.685

Cited by 5 publications

(5 citation statements)

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“…Further, the use of RT-PCR for detection of viral RNA in these studies could not rigorously rule out possible contamination by the virus from the serum. Several laboratories have also shown that HCV can infect B-cell (30), T-cell (18,32,39), and hepatoma cell (14,41) lines in culture, but the infection is usually transient and inefficient. Nevertheless, these studies suggested that B or T cells could support HCV replication, albeit inefficiently, at least in vitro.…”

mentioning

confidence: 99%

Establishment of B-Cell Lymphoma Cell Lines Persistently Infected with Hepatitis C Virus In Vivo and In Vitro: the Apoptotic Effects of Virus Infection

Sung

Shimodaira²,

Doughty

et al. 2003

J Virol

237

181

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Hepatitis C virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Studies of HCV replication and pathogenesis have so far been hampered by the lack of an efficient tissue culture system for propagating HCV in vitro. Although HCV is primarily a hepatotropic virus, an increasing body of evidence suggests that HCV also replicates in extrahepatic tissues in natural infection. In this study, we established a B-cell line (SB) from an HCV-infected non-Hodgkin's B-cell lymphoma. HCV RNA and proteins were detectable by RNase protection assay and immunoblotting. The cell line continuously produces infectious HCV virions in culture. The virus particles produced from the culture had a buoyant density of 1.13 to 1.15 g/ml in sucrose and could infect primary human hepatocytes, peripheral blood mononuclear cells Hepatitis C virus (HCV) has been the major etiological agent of posttransfusion non-A, non-B hepatitis and currently afflicts Ͼ100 million people worldwide. Acute HCV infection is usually subclinical without obvious symptoms. About 15 to 20% of patients can mount a successful immune response to clear the virus in the acute phase; however, 80 to 85% of patients become chronic carriers, and these patients are at high risk of developing liver cirrhosis and/or hepatocellular carcinoma.Besides causing liver pathology, HCV infection is frequently associated with mixed cryoglobulinemia, non-Hodgkin's B-cell lymphoma, and Sjögren's syndrome, all of which involve B-cell proliferation (8, 10, 27, 37, 49; P. Pioltelli, G. Zehender, G. Minti, A. Monteverde, and M. Galli, Letter, Lancet 347:624-625, 1996), suggesting that HCV may infect B cells or affect B-cell functions in natural infection. Negative-strand HCV RNA has been detected by reverse transcriptase (RT) PCR in the peripheral lymphocytes, bone marrow, lymph nodes, and central nervous system of some HCV patients (23,30,34). Analysis of positive-strand HCV RNA sequences and quasispecies patterns suggested that HCV RNAs in these cells are different from those in the serum (22). However, the possibility that HCV replicates in extrahepatic cells remains controversial because of the lack of isolation and characterization of viruses from the infected cells. Further, the use of RT-PCR for detection of viral RNA in these studies could not rigorously rule out possible contamination by the virus from the serum. Several laboratories have also shown that HCV can infect B-cell (30), T-cell (18, 32, 39), and hepatoma cell (14, 41) lines in culture, but the infection is usually transient and inefficient. Nevertheless, these studies suggested that B or T cells could support HCV replication, albeit inefficiently, at least in vitro.The molecular cloning of the HCV genome has made possible the delineation of the gene functions and the potential mechanism of pathogenesis of this virus. Recently, establishment of self-replicating HCV subgenomic (2, 25) and genomic (13, 33) replicons in Huh-7 cells has also provided an important new tool for the study...

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mentioning

confidence: 99%

Establishment of B-Cell Lymphoma Cell Lines Persistently Infected with Hepatitis C Virus In Vivo and In Vitro: the Apoptotic Effects of Virus Infection

Sung

Shimodaira²,

Doughty

et al. 2003

J Virol

237

181

View full text Add to dashboard Cite

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“…Infected Huh7.5 cells were able to shed virus into culture medium, where the later could efficiently infect naïve Huh7.5 cells (results not shown). Infection of cells in vitro with HCV from a variety of sources can at least demonstrate the infectivity of the virus making these systems useful in evaluating drugs for antiviral activity or inhibition of HCV replication [19,22,32]. Transfection with synthetic full-length or near-full-length HCV RNA has the drawback that the RNA used for transfection is homogenously synthesized containing well defined sequence whereas in cases of natural infection with the whole viral particles several quasispecies of different magnitudes of infectivity could be coexisting, thus mimicking the in vivo infections of HCV.…”

Section: Discussionmentioning

confidence: 99%

“…Huh7.5 cells were used for HCV infection experiments and assessment of viral replication. Huh7.50 cells were cultured and infected with HCV particles according to protocols described by Seipp et al, [21] and Song et al,[22]. Assessment of the successful viral infection in Huh7.5 cells throughout the culture duration was confirmed at the transcriptional level by Reverse Transcription Polymerase Chain Reaction (RT-PCR) amplification of HCV RNA and at the translational level by SDS-PAGE of cell lysates followed by Western blotting to determine the de novo synthesis of non structural proteins e.g NS5A.…”

Section: Methodsmentioning

confidence: 99%

Neutralizing activities of caprine antibodies towards conserved regions of the HCV envelope glycoprotein E2

Tabll

Din

et al. 2011

Virol J

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Anti HCV vaccine is not currently available and the present antiviral therapies fail to cure approximately half of the treated HCV patients. This study was designed to assess the immunogenic properties of genetically conserved peptides derived from the C-terminal region of HVR-1 and test their neutralizing activities in a step towards developing therapeutic and/or prophylactic immunogens against HCV infection. Antibodies were generated by vaccination of goats with synthetic peptides derived from HCV E2. Viral neutralizing capacity of the generated anti E2 antibodies was tested using in vitro assays. Goats immunized with E2 synthetic peptides termed p412 [a.a 412-419], p430 [a.a 430-447] and p517 [a.a 517-531] generated high titers of antibody responses 2 to 4.5 fold higher than comparable titers of antibodies to the same epitopes in chronic HCV patients. In post infection experiments of native HCV into cultured Huh7.5 cells anti p412 and anti p 517 were proven to be neutralizing to HCV genotype 4a from patients' sera (87.5% and 75% respectively). On the contrary anti p430 exhibited weak viral neutralization capacity on the same samples (31.25%). Furthermore Ab mixes containing anti p430 exhibited reduced viral neutralization properties. From these experiments one could predict that neutralization by Abs towards different E2-epitopes varies considerably and success in the enrichment of neutralization epitope-specific antibodies may be accompanied by favorable results in combating HCV infection. Also, E2 conserved peptides p517 and p412 represent potential components of a candidate peptide vaccine against HCV infection.

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“…HCV RNA is detected for at least three months following infection. This result also suggested that if the HCV-infected hepatoma cells were co culture with PBMCs, they were able to transfer the virus into PBMCs [16]. …”

Section: Hcv Infection Systemmentioning

confidence: 99%

In-vitro model systems to study Hepatitis C Virus

Ashfaq

Khan

Nawaz

et al. 2011

Genet Vaccines Ther

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Hepatitis C virus (HCV) is a major cause of chronic liver diseases including steatosis, cirrhosis and hepatocellular carcinoma. Currently, there is no vaccine available for prevention of HCV infection due to high degree of strain variation. The current treatment of care, Pegylated interferon α in combination with ribavirin is costly, has significant side effects and fails to cure about half of all infections. The development of in-vitro models such as HCV infection system, HCV sub-genomic replicon, HCV producing pseudoparticles (HCVpp) and infectious HCV virion provide an important tool to develop new antiviral drugs of different targets against HCV. These models also play an important role to study virus lifecycle such as virus entry, endocytosis, replication, release and HCV induced pathogenesis. This review summarizes the most important in-vitro models currently used to study future HCV research as well as drug design.

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Hepatitis C virus infection of human hepatoma cell line 7721 in vitro

Cited by 5 publications

References 0 publications

Establishment of B-Cell Lymphoma Cell Lines Persistently Infected with Hepatitis C Virus In Vivo and In Vitro: the Apoptotic Effects of Virus Infection

Establishment of B-Cell Lymphoma Cell Lines Persistently Infected with Hepatitis C Virus In Vivo and In Vitro: the Apoptotic Effects of Virus Infection

Neutralizing activities of caprine antibodies towards conserved regions of the HCV envelope glycoprotein E2

In-vitro model systems to study Hepatitis C Virus

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