Affiliation:
1. Avid Therapeutics, Inc., Philadelphia, Pennsylvania 19104, USA.
Abstract
We report the development and isolation of a cell line, termed HepAD38, that replicates human hepatitis B virus (HBV) under conditions that can be regulated with tetracycline. In the presence of the antibiotic, this cell line is free of virus due to the repression of pregenomic (pg) RNA synthesis. Upon removal of tetracycline from the culture medium, the cells express viral pg RNA, accumulate subviral particles in the cytoplasm that contain DNA intermediates characteristic of viral replication, and secrete virus-like particles into the supernatant. Since the HepAD38 cell line can produce high levels of HBV DNA, it should be useful for analyses of the viral replication cycle that depend upon viral DNA synthesis in a synchronized fashion. In addition, this cell line has been formatted into a high-throughput, cell-based assay that permits the large-scale screening of diverse compound libraries for new classes of inhibitors of HBV replication.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Pharmacology (medical),Pharmacology
Reference34 articles.
1. Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees;Acs G.;Proc. Natl. Acad. Sci. USA,1987
2. Beasley R. P. and L. Y. Hwang. 1984. Epidemiology of hepatocellular carcinoma p. 209-224. In G. N. Vynas J. L. Deinstag and J. H. Hoofnagle (ed.) Viral hepatitis and liver disease. Grune & Stratton New York N.Y.
3. Amplification of expression of hepatitis B surface antigen in 3T3 cells co-transfected with a dominant-acting gene and cloned viral DNA;Christman J. K.;Proc. Natl. Acad. Sci. USA,1982
4. Transactivation of human immunodeficiency virus occurs via a biomodal mechanism;Cullen B. R.;Cell,1986
5. Inhibition of hepatitis B virus in tissue culture by alpha interferon;Davis M. G.;Antimicrob. Agents Chemother.,1994