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Visualisation of the early steps of human hepatitis B virus infection: a colourful approach

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The human Hepatitis B Virus (HBV) is a major human pathogen. Despite the energetic introduction of a vaccine against HBV in the developed world, still worldwide one in twenty people is chronically infected. People living in poverty, particularly in developing countries, are particularly at risk due to lack of access to vaccines and donor organs. Carriers of the human hepatitis B virus are at high risk of developing liver cirrhosis and hepatocellular carcinoma, and worldwide more than one million people die each year of these diseases. Due to the lack of a small animal model for HBV, and the extreme difficulty until now of in vitro culture of HBV, very little is known about the early stages of HBV entry into human hepatocytes. This proposed programme aims to fill this gap in knowledge through visualisation for the first time of HBV binding and uptake. This will be achieved through the use of a newly-developed cell line, HepaRG, which is permissive to HBV infection, in combination with cutting edge cell biological techniques in the immunofluorescence and confocal microscopy fields. New approaches will include the use of "coloured" viral particles - those containing EGFP or DsRed2 for the visualisation of HBV entry as well as interference assays using HBV peptides anchored in the host cell membrane. In this way, not only will valuable information on HBV entry be gained, but also an in vitro system for the analysis of agents interfering with infection will have been established.

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FP6-2002-MOBILITY-7
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UNIVERSITAETSKLINIKUM HEIDELBERG
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