MassIVE MSV000080718

Imported Reanalysis Dataset Public PXD004724

Label-free Proteomic Analysis of Exosomes Derived from Inducible Hepatitis B Virus-Replicating HepAD38 Cell Line

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Description

Hepatitis B virus (HBV) infection is a major health problem worldwide. Recent evidence suggests that various viruses can manipulate the infection process by secretion of specific viral and cellular components into exosomes, small nanometer-sized (30-150 nm) vesicles secreted from various cells. However, the impact of HBV replication on hepatocytes produced exosomes has not been fully delineated. In this work, an HBV-inducible cell line HepAD38 was used to directly compare changes in the protein content of exosomes secreted from HepAD38 cells with or without HBV replication. Exosomes were isolated from conditioned medium of HepAD38 cell cultures and the purity of exosomes were confirmed by transmission electron microscopy (TEM) and Western immunoblotting assays. Ion-intensity based label-free LC?MS/MS quantitation technologies were applied to analyze protein content of HBV-exosomes and HBV-free-exosomes. A total of 1412 exosomal proteins were identified, in which the abundance of 35 proteins were significantly altered. Strikingly, 5 subunit proteins from the 26S proteasome complex, including PSMC1, PSMC2, PSMD1, PSMD7 and PSMD14 were consistently enhanced in HBV-exosomes. Bioinformatic analysis of differential exosomal proteins revealed the significant enrichment of components involved in proteasomal catabolic process. Proteasome activity assays further suggested that HBV-exosomes had enhanced proteolytic activity compared to HBV-free-exosomes. Furthermore, Human peripheral monocytes incubated with HBV-exosomes induced a significant lower level of IL-6 secretion compared to HBV-free-exosomes. Irreversible inhibition of proteasomal activity within exosome restored the IL-6 induction in monocytes. These results suggest that transmission of proteasome subunit proteins by HBV-exosomes might modulate the innate sensing of pro-inflammatory molecules in the recipient monocytes. These results revealed the composition and potential function of exosomes produced during HBV replication, thus provide a new perspective on the role of exosomes in HBV-host interaction. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: HBV ; Exosomes ; Hepatocyte ; Proteasome ; Label-free quantification

Contact

Principal Investigators:
(in alphabetical order) 		Zhenghong Yuan, Key Laboratory Medical molecular Virology, MoE/MoH Shanghai Medical College, Fudan University 138 Yi Xue Yuan Road, Shanghai, China, 200032, N/A
Submitting User: ccms

Publications

Jia X, Chen J, Megger DA, Zhang X, Kozlowski M, Zhang L, Fang Z, Li J, Chu Q, Wu M, Li Y, Sitek B, Yuan Z.
Label-free Proteomic Analysis of Exosomes Derived from Inducible Hepatitis B Virus-Replicating HepAD38 Cell Line.
Mol. Cell Proteomics. Epub 2017 Mar 27.

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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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