An in vifro System for Infection with Human Hepatitis B Virus Using Tissue Culture System

Park, Sung Soo

Abstract

Hepatits B virus[HBV] is a world wide-pathogen that causes hepatitis. Infection with HBV may be chronic and is associated with subsequent development of hepatocellular carcinoma. In korea the prevalence rate of B hepatitis is estimated to be 7-10%. Despite the severiority of B hepatitis its pathogenesis, chronicity mechanism, prevention and treatment mode have not been developed. The most vexing technical problem in the biology of HBV has been lack of a practical in vitro system that allows infection and replication of this virus under controlled experimental condition. This study was carried out to develop in vitro model of HBV using liver organ culture system. Infection was carried out co-cultivation of 260 micrometer thick-rat liver slices with HBV obtaind from liver biopsy of patient with HBs antigenemia. Aliquots of culture medium were taken in at 6 hour intervals. Hepatitis surface antigen of culture medium in 12 hour incubation increased ab about two folds in comparing with 6 hour incubation. Dot blot analysis of extrachromosomal HBV DNA in cultured rat liver slices using whole HBV DNA as a probe. Fold increment of viral DNA after 12 hours incubation versus 6 hours was measured as 1.7-fold. The presence of viral antigens and Dane particles were observed with immunohistochemical and electron microscopic techniques. In coclusion I present evidece of development for in vitro model of HBV using organ culture system. This system will be useful for understanding the pathogenesis.