Pathogenese der HCV-Infektion: In-vitro-Replikation des Hepatitis-C-Virus und seine genetische Variabilitaet Schlussbericht

1. In spite of the rapid progress in molecular biology of hepatitis C virus (HCV), a major causative agent of chronic liver diseases, the biological characteristics of HCV and the pathogenesis of persistent infection remain obscure. The development of an in vitro system for propagation of HCV is impereative for further investigations into the mechanism of replication, the virus receptor, the morphology of virus particle or the function of the different virus proteins. Considerable genetic heterogeneity has been reported among isolates from different individuals and within the same individual. All known HCV sequences can be classified into at least six major genotypes with a series of subtypes. 2. It is the aim of the present research project to examine cell culture systems for their suitability for HCV replication. Consideration will be given to the existing variety of HCV variants (genotypes and subtypes). The extent of antigen variation due to mutation or selection from a subpopulation of HCV variants was determined. 3. To investigate viral replication system and genomic heterogeneity of HCV we tested different cell lines for their ability to support HCV replication in vitro in connection with PCR and sequence analysis. 4. We have shown - that the human lymphoid cell line H9 and the porcine embryonal kidney cell line POEK supported partial replication of HCV, - that HCV contaminated anti-D immunoglobulin or HCV positive sera could be used to infect culture cells, - that significant sequence variations including geonotype selection detected during propagation of HCV suggest replication of HCV. 5. Provision of a stable in-vitro system for the replication of HCV on a cell culture basis would mean an essential milestone on the route towards elucidation of the pathogenesis of HCV infection. In the present time all cell culture systems, tested are insufficiently with regard of HCV virus replication (world wide). (orig.)SIGLEAvailable from TIB Hannover: DtF QN1(58,59) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Bildung, Wissenschaft, Forschung und Technologie, Bonn (Germany)DEGerman