Novel model of HBV infection and a technology for antiviral agent evaluation
Project is carried out within the European Regional Development Fund (ERDF) activity "Support for science and research"Project identification number: 2010/0233/2DP/188.8.131.52.0/10/APIA/VIAA/075 Project activity period: 36 months (01.01.2011.-31.12.2013.) Principle investigator: Dr. biol. T. Kozlovska
Hepatitis B virus (HBV) is a hepatotrophic virus with a narrow host range, which infects in vitro only primary hepatocytes of humans, chimpanzees and Northern tree shrews (Tupaia belangeri). Availability of those hepatocytes is limited, and the quality of hepatocytes is heterogenous. Therefore easily available and effective HBV infection system is required, which could be used for in vitro screening of antivirals in presence of HBV productive replication and evaluation of HBV neutralizing potential of novel vaccine tools.
Stem cells obtained from adipose tissue (adipose tissue derved stem cells - ADSC) proliferate efficiently, and may give rise to hepatocyte-like cells. The aim of the study is to develop an efficient HBV infection system based on hepatocyte-like cells obtained from ADSC. The hepatocyte-like cells could serve as an efficient HBV infection system for in vitro screening of antivirals and novel vaccines. We have already shown that adipose tissue-derived stem cells which have differentiated to hepatocyte-like cells support infection with HBV.
During antiviral therapy of chronic HBV patients resistant HBV variants emerge, which are also resistant to HBV neutralizing antibodies induced by the current HBV vaccine. The prospective HBV infection model might offer a possibility to evaluate replication efficiency of HBV isolated from chronic HBV patients in presence of antivirals or inteferons, and to develop an individual treatment strategy. This would involve cloning of patient-derived HBV genomes in alphavirus vectors followed by production of recombinant alphaviruses. Infection of hepatocyte-like cells with recombinant alphaviruses would deliver the patient-derived HBV genome variants into hepatocyte-like cells and would allow to monitor the replication of these HBV genome variants. Alphavirus vector-driven initiation of HBV replication has been shown in BHK-21 and HepG2 cells.