Hepatitis B virus is inhibited by RNA interference in cell culture and in mice

• Published in: Antiviral research 2007-01, Vol.73 (1), p.24-30
• Main Authors: Ying, Ruo-Su, Zhu, Cai, Fan, Xue-Gong, Li, Ning, Tian, Xue-Fei, Liu, Hong-Bo, Zhang, Bao-Xin
• Format: Article
• Language: English
• Subjects: Animal model; Animals; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Biological and medical sciences; Cell Line, Tumor; DNA, Viral - metabolism; Female; HBV; Hepatitis B - virology; Hepatitis B Antigens - genetics; Hepatitis B Antigens - metabolism; Hepatitis B virus; Hepatitis B virus - drug effects; HepG2.2.15; Human viral diseases; Humans; Hydrodynamics; Infectious diseases; Medical sciences; Mice; Mice, Inbred BALB C; Pharmacology. Drug treatments; RNA Interference; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; RNA, Viral - genetics; RNA, Viral - metabolism; RNAi; Transfection; Viral diseases; Viral hepatitis; Virus Replication
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2006.05.022

For chronic hepatitis B virus (HBV) infection the effects of current therapies are limited. Recently, RNA interference (RNAi) of virus-specific genes has emerged as a potential antiviral mechanism. Here we studied the effects of HBV-specific 21-bp short hairpin RNAs (shRNAs) targeted to the surface antigen (HBsAg) region and the core antigen (HBcAg) region both in a cell culture system and in a mouse model for HBV replication. HBsAg and hepatitis B e antigen (HBeAg) in the media of the cells and in the sera of the mice were analyzed by time-resolved immunofluorometric assay, intracellular HBcAg by immunofluorescence assay, HBsAg and HBcAg in the livers of the mice by immunohistochemical assay, HBV DNA by fluorogenic quantitative polymerase chain reaction (FQ-PCR) and HBV mRNA by semi-quantitative reverse transcriptase PCR (RT-PCR). Transfection with the shRNAs induced an RNAi response. Secreted HBsAg was reduced by >80% in cell culture and >90% in mouse serum, and HBeAg was also significantly inhibited. Immunofluorescence detection of intracellular HBcAg revealed 76% reduction. In the liver tissues by immunohistochemical detection, there were no HBsAg-positive cells and >70% reduction of HBcAg-positive cells for shRNA-1. And for shRNA-2 the detection of HBsAg and HBcAg also revealed substantial reduction. The shRNAs caused a significant inhibition in the levels of viral mRNA relative to the controls. HBV DNA was reduced by >40% for shRNA-1 and >60% for shRNA-2. RNAi is capable of inhibiting HBV replication and expression in vitro and in vivo and thus may constitute a new therapeutic strategy for HBV infection.