Different Requirements for Scavenger Receptor Class B Type I in Hepatitis C Virus Cell-Free versus Cell-to-Cell Transmission

Catanese, Maria Teresa; Loureiro, Joana; Jones, Christopher T.; Dorner, Marcus; Hahn, Thomas von; Rice, Charles M.

doi:10.1128/jvi.01102-13

Cited by 77 publications

(66 citation statements)

References 58 publications

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“…AR4A potently neutralizes the virus used in this study and displays one of the highest inhibitory effects among a panel of α-HCV antibodies screened (16,17), arguing against insufficient binding properties (Fig. 1).…”

Section: Resultsmentioning

confidence: 93%

Ultrastructural analysis of hepatitis C virus particles

Catanese

Uryu

Kopp

et al. 2013

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

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Hepatitis C virus (HCV) is a major cause of chronic liver disease, with an estimated 170 million people infected worldwide. Low yields, poor stability, and inefficient binding to conventional EM grids have posed significant challenges to the purification and structural analysis of HCV. In this report, we generated an infectious HCV genome with an affinity tag fused to the E2 envelope glycoprotein. Using affinity grids, previously described to isolate proteins and macromolecular complexes for single-particle EM, we were able to purify enveloped particles directly from cell culture media. This approach allowed for rapid in situ purification of virions and increased particle density that were instrumental for cryo-EM and cryoelectron tomography (cryo-ET). Moreover, it enabled ultrastructural analysis of virions produced by primary human hepatocytes. HCV appears to be the most structurally irregular member of the Flaviviridae family. Particles are spherical, with spike-like projections, and heterogeneous in size ranging from 40 to 100 nm in diameter. Exosomes, although isolated from unfractionated culture media, were absent in highly infectious, purified virus preparations. Cryo-ET studies provided low-resolution 3D structural information of highly infectious virions. In addition to apolipoprotein (apo)E, HCV particles also incorporate apoB and apoA-I. In general, host apolipoproteins were more readily accessible to antibody labeling than HCV glycoproteins, suggesting either lower abundance or masking by host proteins.enveloped virus | hepacivirus | lipoviral particle | virus structure | virus assembly H epatitis C virus (HCV) is an important human pathogen that infects the liver and establishes chronic infection in the majority of cases, leading to cirrhosis and hepatocellular carcinoma (HCC) over the course of many years. More than 170 million people, ∼3% of the world's population, have been infected with HCV. Each year, 4-5% of patients with HCVinduced cirrhosis develop HCC, making HCV infection the leading indicator for liver transplantation in many areas of the world (1). Surgery, however, does not provide a cure because the donor organ universally becomes reinfected. A prophylactic vaccine is not available and despite the recent addition of HCV-specific protease inhibitors to the pegylated (peg)-IFN and ribavirin regimen, which has increased the cure rate, better therapies are still needed to solve the emergence of resistant variants, severe side effects and suboptimal response rates in cirrhotic patients (2).HCV is a single-stranded, positive-sense RNA virus in the family Flaviviridae. The HCV genome is ∼9.6 kb in length and encodes a long polyprotein of more than 3000 amino acids that is proteolytically processed to generate 10 mature viral proteins. Viral structural proteins are encoded by the first third of the polyprotein and include core or capsid protein (C) and the envelope glycoproteins E1 and E2. p7 (a viroporin) and nonstructural proteins, encoded by the C-terminal two-thirds of the polyprotein, pla...

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Section: Resultsmentioning

confidence: 93%

Ultrastructural analysis of hepatitis C virus particles

Catanese

Uryu

Kopp

et al. 2013

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

Self Cite

250

238

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“…In contrast, HCV cell-to-cell transmission is less well defined due to controversial observations. Besides the disagreement on the role of CD81 in HCV cell-to-cell transmission (45,46), the requirements for complete HCV assembly machinery (from both virus and host sides) are under debate.…”

Section: Discussionmentioning

confidence: 99%

Visualizing the Essential Role of Complete Virion Assembly Machinery in Efficient Hepatitis C Virus Cell-to-Cell Transmission by a Viral Infection-Activated Split-Intein-Mediated Reporter System

Zhao

Deng

et al. 2017

J Virol

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Hepatitis C virus (HCV) infects 2 to 3% of the world population and is a leading cause of liver diseases such as fibrosis, cirrhosis, and hepatocellular carcinoma. Many aspects of HCV study, ranging from molecular virology and antiviral drug development to drug resistance profiling, were supported by straightforward assays of HCV replication and infection. Among these assays, the HCV-dependent fluorescence relocalization (HDFR) system allowed live-cell visualization of infection without modifying the viral genome, but this strategy required careful recognition of the fluorescence relocalization pattern for its high fluorescence background in the cytoplasm. In this study, to achieve background-free visualization of HCV infection, a viral infection-activated split-inteinmediated reporter system (VISI) was devised. Uninfected Huh7.5.1-VISI cells show no background signal, while HCV infection specifically illuminates the nuclei of infected Huh7.5.1-VISI cells with either green fluorescent protein (GFP) or mCherry. Combining VISI-GFP and VISI-mCherry systems, we revisited HCV cell-to-cell transmission with clearcut distinction of donor and recipient cells in a live-cell manner. Independently of virion assembly, exosomes have been reported to transfer HCV subgenomic RNA to initiate replication in uninfected cells, which suggested an assembly-free pathway. However, our data demonstrated that HCV structural genes and the p7 gene were essential for not only cell-free infectivity but also cell-to-cell transmission. Additionally, depletion of apolipoprotein E (ApoE) from donor cells but not from recipient cells significantly reduced HCV cell-to-cell transmission efficiency. In summary, we developed a background-free cell-based reporter system for convenient live-cell visualization of HCV infection, and our data indicate that complete HCV virion assembly machinery is essential for both cell-free and cell-to-cell transmission. IMPORTANCE Hepatitis C virus (HCV) infects hepatocytes via two pathways: cell-free infection and cell-to-cell transmission. Structural modules of the HCV genome are required for production of infectious cell-free virions; however, the role of specific genes within the structural module in cell-to-cell transmission is not clearly defined. Our data demonstrate that deletion of core, E1E2, and p7 genes individually results in no HCV cell-to-cell transmission and that ApoE knockdown from donor cells causes less-efficient cell-to-cell transmission. Thus, this work indicates that the complete HCV assembly machinery is required for HCV cell-to-cell transmission. At last, this work presents an optimized viral infection-activated split-intein-mediated reporter system for easy live-cell monitoring of HCV infection.

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“…Apical secretion of measles virus particles is inefficient (Leonard et al, 2008), but infectious centers with many dying cells that detach from the epithelium may also be expulsed by coughing and sneezing. in absence of cell-free virus particles mainly relies on claudin-1 (Timpe et al, 2008) through a mechanism that might require -yet unknown -co-factors (Catanese et al, 2013). The recent development of primary liver cultures should help to answer important questions regarding the cell-to-cell spread of HCV in relevant model systems (Andrus et al, 2011;Sourisseau et al, 2013).…”

Section: Tjs -Components and Interactionsmentioning

confidence: 99%

Connections matter − how viruses use cell–cell adhesion components

Mateo

Generous

Sinn

et al. 2015

Journal of Cell Science

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The epithelium is a highly organized type of animal tissue. Except for blood and lymph vessels, epithelial cells cover the body, line its cavities in single or stratified layers and support exchange between compartments. In addition, epithelia offer to the body a barrier to pathogen invasion. To transit through or to replicate in epithelia, viruses have to face several obstacles, starting from cilia and glycocalyx where they can be neutralized by secreted immunoglobulins. Tight junctions and adherens junctions also prevent viruses to cross the epithelial barrier. However, viruses have developed multiple strategies to blaze their path through the epithelium by utilizing components of cell–cell adhesion structures as receptors. In this Commentary, we discuss how viruses take advantage of the apical junction complex to spread. Whereas some viruses quickly disrupt epithelium integrity, others carefully preserve it and use cell adhesion proteins and their cytoskeletal connections to rapidly spread laterally. This is exemplified by the hidden transmission of enveloped viruses that use nectins as receptors. Finally, several viruses that replicate preferentially in cancer cells are currently used as experimental cancer therapeutics. Remarkably, these viruses use cell adhesion molecules as receptors, probably because – to reach tumors and metastases – oncolytic viruses must efficiently traverse or break epithelia.

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Different Requirements for Scavenger Receptor Class B Type I in Hepatitis C Virus Cell-Free versus Cell-to-Cell Transmission

Cited by 77 publications

References 58 publications

Ultrastructural analysis of hepatitis C virus particles

Ultrastructural analysis of hepatitis C virus particles

Visualizing the Essential Role of Complete Virion Assembly Machinery in Efficient Hepatitis C Virus Cell-to-Cell Transmission by a Viral Infection-Activated Split-Intein-Mediated Reporter System

Connections matter − how viruses use cell–cell adhesion components

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