Development and application of an in vitro model for screening anti-hepatitis B virus therapeutics

✍ Scribed by Pietro Lampertico; James S. Malter; Michael A. Gerber

• Publisher: John Wiley and Sons
• Year: 1991
• Tongue: English
• Weight: 592 KB
• Volume: 13
• Category: Article
• ISSN: 0270-9139
• DOI: 10.1002/hep.1840130307

✦ Synopsis

The development of effective anti-hepatitis B virus agents has been hampered by the lack of reliable in uitro systems for the screening of new therapeutics. In an effort to circumvent this problem, we have developed an in uitro system for screening antihepatitis B virus drugs using hepatitis B virus DNA-transfectedHep G2 cells. The cell line designated 2.2.15 produces replicative viral DNA intermediates, mature Dane particles and high levels of viral antigens. Subconfluent 2.2.15 cells were treated with a variety of commonly used anti-hepatitis B virus therapeutics, and their efficacy was determined by analyzing changes in the replicative cellular or extracellular hepatitis B virus DNA content by Southern blotting or slot-blot hybridization. The slot-blot method was sensitive, reproducible and rapid and correlated well with Southern blotting. Analysis of the media for hepatitis B virus DNA was indicative of changes in intracellular, replicative hepatitis B virus DNA, permitting sampling of the media. Therefore 2.2.15 cells may provide a valuable method for identifying and monitoring effective anti-hepatitis B virus therapeutics. Using this system to test various agents, we confirm that 2'-deoxyguanosine strongly inhibited viral replication, whereas others tested were less effective. Correlation with in uiuo systems is now needed. (HEPATOLOGY 1991; 13~422-426.) Chronic infection with HBV is often associated with severe, persistent liver disease that ranges from CAH to cirrhosis and HCC (1-4). Despite the worldwide magnitude of chronic HBV infection, effective antiviral agents remain unavailable. A variety of drugs, including adenine arabinoside (ARA-A), adenine arabinoside monophosphate (ARA-AMP) and interferons (IFNs), have displayed only partial or transient anti-HBV activity, often with deleterious side effects (5-9).

The development and study of new anti-HBV drugs have been hampered. by the lack of reliable in vitro models that support HBV replication. Long-term cul-