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Abstract
The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enhanced by treating cells with polyethylene glycol 8000 (PEG) during infection. However, this alone does not promote virus spread. Here we show that maintaining PEG in culture medium increases the rate of infection by at least one order of magnitude, and, most importantly, promotes virus spread. To demonstrate the utility of this system, we show that two interferon-stimulated genes (ISGs), ISG20 and tetherin, restrict HBV spread in NTCP-expressing hepatoma cells. Thus, this protocol can be easily applied to existing cell culture systems to study the complete HBV life cycle, including virus spread.
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Details
1 Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, USA
2 Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, USA; Department of Microbiology and Center of Infectious Disease, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China
3 Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA
4 Institute for Medical Engineering and Science, David H. Koch Institute for Integrative Cancer Research, Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA, USA; Howard Hughes Medical Institute, Cambridge, MA, USA
5 Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, USA; Division of Gastroenterology and Hepatology, Weill Cornell Medical College, New York, NY, USA
6 Department of Medicine D and the Liver Institute, Rabin Medical Center, Belinson Hospital, Petach-Tikva and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel