Exosome-associated hepatitis C virus in cell cultures and patient plasma

Liu, Ziqing; Zhang, Xiugen; Yu, Qigui; He, Johnny J.

doi:10.1016/j.bbrc.2014.10.146

Cited by 87 publications

(75 citation statements)

References 27 publications

(32 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A link between exosomes and HCV was first suggested by electron microscopy analysis of HCV virion samples for the similarity in morphology of exosomes and observed virion structure (7). Exosomes prepared by ultracentrifugation were further reported to transmit HCV infection productively (49). However, these observations are not conclusive as morphological and biophysical characteristics of exosomes are very comparable to those of cell-free infectious HCV LVP.…”

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

View full text Add to dashboard Cite

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.

show abstract

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

View full text Add to dashboard Cite

show abstract

“…Previous studies on serum samples showed that exosomes contained virus-encoded miRNAs (EBV, KSHV, JCV, and HCV) [15, 23–25]. However, the detailed profiles of virus-encoded miRNAs in both exosomes and intracellularly have not been reported yet.…”

Section: Discussionmentioning

confidence: 99%

Profile of Exosomal and Intracellular microRNA in Gamma-Herpesvirus-Infected Lymphoma Cell Lines

et al. 2016

View full text Add to dashboard Cite

Exosomes are small vesicles released from cells, into which microRNAs (miRNA) are specifically sorted and accumulated. Two gamma-herpesviruses, Kaposi sarcoma-associated herpesvirus (KSHV) and Epstein—Barr virus (EBV), encode miRNAs in their genomes and express virus-encoded miRNAs in cells and exosomes. However, there is little information about the detailed distribution of virus-encoded miRNAs in cells and exosomes. In this study, we thus identified virus- and host-encoded miRNAs in exosomes released from KSHV- or EBV-infected lymphoma cell lines and compared them with intracellular miRNAs using a next-generation sequencer. Sequencing analysis demonstrated that 48% of the annotated miRNAs in the exosomes from KSHV-infected cells originated from KSHV. Human mir-10b-5p and mir-143-3p were much more highly concentrated in exosomes than in cells. Exosomes contained more nonexact mature miRNAs that did not exactly match those in miRBase than cells. Among the KSHV-encoded miRNAs, miRK12-3-5p was the most abundant exact mature miRNA in both cells and exosomes that exactly matched those in miRBase. Recently identified EXOmotifs, nucleotide motifs that control the loading of miRNAs into exosomes were frequently found within the sequences of KSHV-encoded miRNAs, and the presence of the EXOmotif CCCT or CCCG was associated with the localization of miRNA in exosomes in KSHV-infected cells. These observations suggest that specific virus-encoded miRNAs are sorted by EXOmotifs and accumulate in exosomes in virus-infected cells.

show abstract

“…Extracellular vesicles (EVs), including microvesicles (MVs), or exosomes, have been shown to serve as vehicles for intercellular communication and transfer of genetic material . Several studies suggest the potential role of extracellular vesicles and their biogenesis pathways in HCV and HBV infection, via exosomes . Evidence suggests that hepatitis B virus (HBV) infects its target cells in the form of cell‐free viruses or through cell‐cell contact.…”

Section: Introductionmentioning

confidence: 99%

Extracellular vesicles from hepatitis B patients serve as reservoir of hepatitis B virus DNA

Sukriti

Choudhary

Maras

et al. 2018

Journal of Viral Hepatitis

View full text Add to dashboard Cite

Exosome-associated hepatitis C virus in cell cultures and patient plasma

Cited by 87 publications

References 27 publications

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

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

Profile of Exosomal and Intracellular microRNA in Gamma-Herpesvirus-Infected Lymphoma Cell Lines

Extracellular vesicles from hepatitis B patients serve as reservoir of hepatitis B virus DNA

Contact Info

Product

Resources

About