Proteomic analysis of primary duck hepatocytes infected with duck hepatitis B virus

biomed - Zhao Yanfeng , Ben Haijing , Qu Su , Zhou Xinwen , Yan Liang , Xu Bin , Zhou Shuangcheng , Lou Qiang , Ye Rong , Zhou Tianlun , Yang Pengyuan , Qu Di

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16 pages

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Hepatitis B virus (HBV) is a major cause of liver infection in human. Because of the lack of an appropriate cell culture system for supporting HBV infection efficiently, the cellular and molecular mechanisms of hepadnavirus infection remain incompletely understood. Duck heptatitis B virus (DHBV) can naturally infect primary duck hepatocytes (PDHs) that provide valuable model systems for studying hepadnavirus infection in vitro . In this report, we explored global changes in cellular protein expression in DHBV infected PDHs by two-dimension gel electrophoresis (2-DE) combined with MALDI-TOF/TOF tandem mass spectrometry (MS/MS). Results The effects of hepadnavirus infection on hepatocytes were investigated in DHBV infected PDHs by the 2-DE analysis. Proteomic profile of PDHs infected with DHBV were analyzed at 24, 72 and 120 h post-infection by comparing with uninfected PDHs, and 75 differentially expressed protein spots were revealed by 2-DE analysis. Among the selected protein spots, 51 spots were identified corresponding to 42 proteins by MS/MS analysis; most of them were matched to orthologous proteins of Gallus gallus , Anas platyrhynchos or other avian species, including alpha-enolase, lamin A, aconitase 2, cofilin-2 and annexin A2, etc. The down-regulated expression of beta-actin and annexin A2 was confirmed by Western blot analysis, and potential roles of some differentially expressed proteins in the virus-infected cells have been discussed. Conclusions Differentially expressed proteins of DHBV infected PDHs revealed by 2-DE, are involved in carbohydrate metabolism, amino acid metabolism, stress responses and cytoskeleton processes etc, providing the insight to understanding of interactions between hepadnavirus and hepatocytes and molecular mechanisms of hepadnavirus pathogenesis.

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Publié par	biomed
Publié le	01 janvier 2010
Nombre de lectures	15
Langue	English
Poids de l'ouvrage	1 Mo

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Zhao et al. Proteome Science 2010, 8 :28 http://www.proteomesci.com/content/8/1/28

R E S E A R C H Open Access R P es r e o arc t h eomic analysis of primary duck hepatocytes infected with duck hepatitis B virus Yanfeng Zhao †1 , Haijing Ben †1 , Su Qu †1 , Xinwen Zhou 2 , Liang Yan 1 , Bin Xu 1 , Shuangcheng Zhou 1 , Qiang Lou 1 , Rong Ye 1 , Tianlun Zhou 1 , Pengyuan Yang 2 and Di Qu* 1

Introduction study HBV infection [3]. Human primary hepatocytes The HBV, prototype of the Hepadnaviridae family, is a and HepaRG cells can support HBV life cycle, but have noncytopathic hepatotropic DNA virus replicating via limitations in accessibility, reproducibility and low level reverse transcription [1]. More than 350 million individu- of HBV replication, and a large amount of input virus was als are HBV carriers worldwide and over one-third of needed to infect low proportion of cells [4-6]. DHBV and them develop serious liver diseases such as chronic hepa- woodchuck hepatitis B virus (WHBV) are classified into titis, cirrhosis and primary hepatocellular carcinoma [2]. the family of hepadnaviridae. Thus for hepadnavirus Major obstacles in HBV research have been the inability infection primary hepatocytes of ducks (DHBV) and of the virus to infect cells in vitro and lack of adequate woodchucks (WHBV) are still considered as suitable animal models for HBV infection, though primary human models for investigating the viral replication and patho-hepatocytes and HepaRG cell line have been used to genesis [7,8]. The development of proteomic methods has enabled us * Correspondence: dqu@shmu.edu.cn to investigate the changes of cellular protein expression at 1 Key Laboratory of Medical Molecular Vi rology of Ministries of Education and a global scale to reveal virus-host interactions [9-12]. The Health, Institute of Medical Microbiology and Institutes of Biomedical Sciences, effect of hepadnavirus replication on the host cells, such Shanghai Medical College of Fudan University, Shanghai, China the carcinoma derived hepatocyte lines transfected † Contributed equally as Full list of author information is available at the end of the article with the HBV genome, HepaRG cell lines or HBV trans-© 2010 Zhao et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Comm ons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestri cted use, distribution, and reproduction in any medium, provided the original work is properly cited.