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

Identifieur interne : 000D12 ( Main/Exploration ); précédent : 000D11; suivant : 000D13

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

Auteurs : Yanfeng Zhao [République populaire de Chine] ; Haijing Ben [République populaire de Chine] ; Su Qu [République populaire de Chine] ; Xinwen Zhou [République populaire de Chine] ; Liang Yan [République populaire de Chine] ; Bin Xu [République populaire de Chine] ; Shuangcheng Zhou [République populaire de Chine] ; Qiang Lou [République populaire de Chine] ; Rong Ye [République populaire de Chine] ; Tianlun Zhou [République populaire de Chine] ; Pengyuan Yang [République populaire de Chine] ; Di Qu [République populaire de Chine]

Source :

RBID : PMC:2904733

Abstract

Background

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.


Url:
DOI: 10.1186/1477-5956-8-28
PubMed: 20529248
PubMed Central: 2904733


Affiliations:


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Le document en format XML

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<name sortKey="Lou, Qiang" sort="Lou, Qiang" uniqKey="Lou Q" first="Qiang" last="Lou">Qiang Lou</name>
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<wicri:regionArea>Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, Institute of Medical Microbiology and Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai</wicri:regionArea>
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<name sortKey="Ye, Rong" sort="Ye, Rong" uniqKey="Ye R" first="Rong" last="Ye">Rong Ye</name>
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<name sortKey="Yang, Pengyuan" sort="Yang, Pengyuan" uniqKey="Yang P" first="Pengyuan" last="Yang">Pengyuan Yang</name>
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<title>Background</title>
<p>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
<italic>in vitro</italic>
. 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).</p>
</sec>
<sec>
<title>Results</title>
<p>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
<italic>Gallus gallus</italic>
,
<italic>Anas platyrhynchos </italic>
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.</p>
</sec>
<sec>
<title>Conclusions</title>
<p>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.</p>
</sec>
</div>
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