Permissivity of primary hepatocytes and hepatoma cell lines to support hepatitis C virus infection

Wilson, Garrick K.; Farquhar, Michelle J.; Meredith, Luke W.; Dhawan, Anil; Mitry, Ragai R.; Balfe, Peter; McKeating, Jane A.

doi:10.1099/vir.0.000085

Cited by 3 publications

(3 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While some groups have reported successful culture of patient-derived HCV, these methods generally exhibit poor replication efficiency and therefore do not represent a tractable means of predicting neutralization sensitivity in vivo ; instead, surrogate models, such as the HCVpp system or chimeric HCVcc, have proven to be more robust tools (reviewed in reference 42 ). Previous studies on antibody neutralization have used the cell line Huh7.5 ( 40 ) or primary hepatocytes ( 43 ); we chose the former, as that cell line is more amenable and the HCV entry process is comparable to that observed in primary hepatocytes ( 44 – 46 ).…”

Section: Discussionmentioning

confidence: 99%

A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance

Urbanowicz

McClure

Brown

et al. 2016

J Virol

100

View full text Add to dashboard Cite

Despite significant advances in the treatment of hepatitis C virus (HCV) infection, the need to develop preventative vaccines remains. Identification of the best vaccine candidates and evaluation of their performance in preclinical and clinical development will require appropriate neutralization assays utilizing diverse HCV isolates. We aimed to generate and characterize a panel of HCV E1E2 glycoproteins suitable for subsequent use in vaccine and therapeutic antibody testing. Full-length E1E2 clones were PCR amplified from patient-derived serum samples, cloned into an expression vector, and used to generate viral pseudoparticles (HCVpp). In addition, some of these clones were used to generate cell culture infectious (HCVcc) clones. The infectivity and neutralization sensitivity of these viruses were then determined. Bioinformatic and HCVpp infectivity screening of approximately 900 E1E2 clones resulted in the assembly of a panel of 78 functional E1E2 proteins representing distinct HCV genotypes and different stages of infection. These HCV glycoproteins differed markedly in their sensitivity to neutralizing antibodies. We used this panel to predict antibody efficacy against circulating HCV strains, highlighting the likely reason why some monoclonal antibodies failed in previous clinical trials. This study provides the first objective categorization of cross-genotype patient-derived HCV E1E2 clones according to their sensitivity to antibody neutralization. It has shown that HCV isolates have clearly distinguishable neutralization-sensitive, -resistant, or -intermediate phenotypes, which are independent of genotype. The panel provides a systematic means for characterization of the neutralizing response elicited by candidate vaccines and for defining the therapeutic potential of monoclonal antibodies.IMPORTANCE Hepatitis C virus (HCV) has a global burden of more than 170 million people, many of whom cannot attain the new, expensive, direct-acting antiviral therapies. A safe and effective vaccine that generates both T cell responses and neutralizing antibodies is required to eradicate the disease. Regions within the HCV surface glycoproteins E1 and E2 are essential for virus entry and are targets for neutralizing antibodies. Screening of vaccine candidates requires suitable panels of glycoproteins that represent the breadth of neutralization resistance. Use of a standard reference panel for vaccine studies will ensure comparability of data sets, as has become routine for HIV-1. Here, we describe a large panel of patient-derived HCV glycoproteins with an assessment of their neutralization sensitivity to defined monoclonal antibodies, which has enabled us to predict their likely efficacy in the wider HCV-infected population. The panel could also be important for future selection of additional therapeutic antibodies and for vaccine design.

show abstract

Section: Discussionmentioning

confidence: 99%

A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance

Urbanowicz

McClure

Brown

et al. 2016

J Virol

100

View full text Add to dashboard Cite

show abstract

“…Our understanding of the parameters defining infectious HCV in humans is incomplete due to the limited replication of primary HCV strains in any in vitro model system ( 56 , 57 ). Previous studies have reported on the infectivity of patient-derived sera in chimpanzees ( 34 ) and humanized uPA-SCID mice ( 42 , 54 , 58 ), however, these studies were limited by the small number of clinical samples.…”

Section: Discussionmentioning

confidence: 99%

A Role for B Cells to Transmit Hepatitis C Virus Infection

et al. 2021

View full text Add to dashboard Cite

Hepatitis C virus (HCV) is highly variable and transmits through infected blood to establish a chronic liver infection in the majority of patients. Our knowledge on the infectivity of clinical HCV strains is hampered by the lack of in vitro cell culture systems that support efficient viral replication. We and others have reported that HCV can associate with and infect immune cells and may thereby evade host immune surveillance and elimination. To evaluate whether B cells play a role in HCV transmission, we assessed the ability of B cells and sera from recent (<2 years) or chronic (≥ 2 years) HCV patients to infect humanized liver chimeric mice. HCV was transmitted by B cells from chronic infected patients whereas the sera were non-infectious. In contrast, B cells from recently infected patients failed to transmit HCV to the mice, whereas all serum samples were infectious. We observed an association between circulating anti-glycoprotein E1E2 antibodies and B cell HCV transmission. Taken together, our studies provide evidence for HCV transmission by B cells, findings that have clinical implications for prophylactic and therapeutic antibody-based vaccine design.

show abstract

“…Our understanding of the parameters de ning infectious HCV in man is incomplete due to the limited replication of clinical or primary HCV strains in in vitro model systems (50,51). Previous studies have reported on the infectivity of patient-derived sera in chimpanzees (32) and humanized uPA-SCID mice (39,44,52), however, these studies were limited by the small number of clinical samples studied.…”

Section: Discussionmentioning

confidence: 99%

A role for B cells to transmit hepatitis C virus infection

Desombere

Houtte

Farhoudi

et al. 2020

Preprint

View full text Add to dashboard Cite

Hepatitis C virus (HCV) is highly variable and transmits through infected blood to establish a chronic liver infection in the majority of patients. Our knowledge of the infectivity of clinical HCV strains is hampered by the lack of in vitro cell culture systems that support efficient viral replication. We previously reported that laboratory strains of HCV associated with non-permissive B cells could trans-infect hepatocytes and thereby evade host neutralizing antibody responses, suggesting a role for B cells in HCV transmission. To evaluate this hypothesis, we assessed the ability of B cells and sera from recent (<2 years) or chronic (≥ 2 years) infections to infect humanized liver chimeric mice. HCV was efficiently transmitted by B cells from chronically infected patients whereas the sera were non-infectious. In contrast, we noted that B cells from recently infected patients failed to transmit HCV to the mice, whereas all serum samples were infectious. Only patients with circulating anti-glycoprotein antibodies harbored genomic HCV-RNA in B cells. Taken together, our studies provide direct in vivo evidence for HCV transmission by B cells and these findings may have clinical implications for prophylactic and therapeutic antibody-based vaccine design.

show abstract

Permissivity of primary hepatocytes and hepatoma cell lines to support hepatitis C virus infection

Cited by 3 publications

References 25 publications

A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance

A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance

A Role for B Cells to Transmit Hepatitis C Virus Infection

A role for B cells to transmit hepatitis C virus infection

Contact Info

Product

Resources

About