Expression of E1E2 on Hepatitis C RNA-Containing Particles Released from Primary Cultured Human Hepatocytes Derived from Infected Cirrhotic Livers

Ndongo, Ndiémé; Selliah, Subjini; Berthillon, Pascale; Raymond, Valérie–Ann; Trépo, Christian; Bilodeau, Marc; Petit, Marie-Anne

doi:10.1159/000318886

Cited by 2 publications

(1 citation statement)

References 41 publications

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“…The E1E2-expressing HCV pseudo particles were shown to be excellent HCV models to study viral binding at the cell surface [ 12 ]. HepaRG and liver primary cells have been described as positive models to bind, incorporate and replicate HCV [ 25 , 26 , 36 , 38 ]. The characteristics of several mosquito cell surface receptors have been described, but they remain poorly understood.…”

Section: Discussionmentioning

confidence: 99%

New insights into HCV replication in original cells from Aedes mosquitoes

Fallecker

Caporossi

Rechoum

et al. 2017

Virol J

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BackgroundThe existing literature about HCV association with, and replication in mosquitoes is extremely poor. To fill this gap, we performed cellular investigations aimed at exploring (i) the capacity of HCV E1E2 glycoproteins to bind on Aedes mosquito cells and (ii) the ability of HCV serum particles (HCVsp) to replicate in these cell lines.MethodsFirst, we used purified E1E2 expressing baculovirus-derived HCV pseudo particles (bacHCVpp) so we could investigate their association with mosquito cell lines from Aedes aegypti (Aag-2) and Aedes albopictus (C6/36). We initiated a series of infections of both mosquito cells (Ae aegypti and Ae albopictus) with the HCVsp (Lat strain - genotype 3) and we observed the evolution dynamics of viral populations within cells over the course of infection via next-generation sequencing (NGS) experiments.ResultsOur binding assays revealed bacHCVpp an association with the mosquito cells, at comparable levels obtained with human hepatocytes (HepaRG cells) used as a control. In our infection experiments, the HCV RNA (+) were detectable by RT-PCR in the cells between 21 and 28 days post-infection (p.i.). In human hepatocytes HepaRG and Ae aegypti insect cells, NGS experiments revealed an increase of global viral diversity with a selection for a quasi-species, suggesting a structuration of the population with elimination of deleterious mutations. The evolutionary pattern in Ae albopictus insect cells is different (stability of viral diversity and polymorphism).ConclusionsThese results demonstrate for the first time that natural HCV could really replicate within Aedes mosquitoes, a discovery which may have major consequences for public health as well as in vaccine development.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-017-0828-z) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

New insights into HCV replication in original cells from Aedes mosquitoes

Fallecker

Caporossi

Rechoum

et al. 2017

Virol J

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Long-term propagation of serum hepatitis C virus (HCV) with production of enveloped HCV particles in human HepaRG hepatocytes

Ndongo-Thiam

Berthillon

Errazuriz³

et al. 2011

Hepatology

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HepaRG human liver progenitor cells exhibit morphology and functionality of adult hepatocytes. We investigated the susceptibility of HepaRG hepatocytes to in vitro infection with serum-derived hepatitis C virus (HCV) particles (HCVsp) and the potential neutralizing activity of the E1E2-specific monoclonal antibody (mAb) D32.10. The infection was performed using HCVsp when the cells actively divided at day 3 postplating. HCV RNA, E1E2, and core antigens were quantified in HCV particles recovered from culture supernatants of differentiated cells for up to 66 days. The density distributions of particles were analyzed on iodixanol or sucrose gradients. Electron microscopy (EM) and immune-EM studies were performed for ultrastructural analysis of cells and localization of HCV E1E2 proteins in thin sections. HCV infection of HepaRG cells was documented by increasing production of E1E2-core-RNA(1) HCV particles from day 21 to day 63. Infectious particles sedimented between 1.06 and 1.12 g/mL in iodixanol gradients. E1E2 and core antigens were expressed in 50% of HCV-infected cells at day 31. The D32.10 mAb strongly inhibited HCV RNA production in HepaRG culture supernatants. Infected HepaRG cells frozen at day 56 were reseeded at low density. After only 1-3 subcultures and induction of a cell differentiation process the HepaRG cells produced high titer HCV RNA and thus showed to be sustainably infected. Apolipoprotein B-associated empty E1E2 and complete HCV particles were secreted. Characteristic virus-induced intracellular membrane changes and E1E2 protein-association to vesicles were observed. Conclusion: HepaRG progenitor cells permit HCVsp infection. Differentiated HepaRG cells support longterm production of infectious lipoprotein-associated enveloped HCV particles. The E1E2-specific D32.10 mAb neutralizes the infection and this cellular model could be used as a surrogate infection system for the screening of entry inhibitors. (HEPATOLOGY 2011;54:406-417) H epatitis C virus (HCV) infection is a major health problem worldwide because at least 70% of infections persist and cause chronic hepatitis, which may progress to liver cirrhosis and hepatocellular carcinoma. 1 The lack of robust cell culture and small animal models remain stumbling blocks to

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Expression of E1E2 on Hepatitis C RNA-Containing Particles Released from Primary Cultured Human Hepatocytes Derived from Infected Cirrhotic Livers

Cited by 2 publications

References 41 publications

New insights into HCV replication in original cells from Aedes mosquitoes

New insights into HCV replication in original cells from Aedes mosquitoes

Long-term propagation of serum hepatitis C virus (HCV) with production of enveloped HCV particles in human HepaRG hepatocytes

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