Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 180, Issue 1-2, Pages 84-90Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2011.12.016
Keywords
Hepatitis B virus; Interferon gamma; Gene regulation; pcDNA3.1; Antiviral
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Interferon-gamma (IFN-gamma) has been reported to have antiviral activity against Hepatitis B virus (HBV) and to suppress HBV replication noncytolytically in vivo. Since systemic administration of IFN-gamma may cause severe adverse effects, studies of the effects of liver-specific IFN-gamma expression from adenoviral vectors in vivo have been investigated. In this study, a novel strategy has been described that drives specific expression of human IFN-gamma in HBsAg-secreting hepatocytes. A bicistronic expression vector has been developed, pcDNA3.1-HBV antisense S gene-HCV core protein gene-HCV internal ribosome entry sites (IRES)-IFN-gamma (pcDNA-SCI gamma), by inserting four DNA fragments into pcDNA3.1. Tight modulation of HCV IRES-dependent translation by the HCV core protein was achieved using an antisense RNA technique with a bicistronic expression vector. HepG2 cells and HepG2.2.15 cells stably expressing HBV were transduced with pcDNA-SCI gamma to test the responsiveness of IFN-gamma to HBsAg expression. Gene transfer resulted in a low background and a 30-fold induction of IFN-gamma expression from pcDNA-SCI gamma in a cell-specific fashion. Hepatocyte-specific IFN-gamma expression controlled effectively HBV replication in HBsAg-secreting HepG2.2.15 cells without cell toxicity. (C) 2012 Elsevier B.V. All rights reserved.
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