Robust in vitro HBV infection systemsregist2.virology-education.com/presentations/2017/... · 2017....
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Robust in vitro HBV infection systems Lefteris Michailidis, Ph.D. Laboratory of Virology and Infectious Disease Charles M. Rice Laboratory The Rockefeller University December 4, 2017
Transcript of Robust in vitro HBV infection systemsregist2.virology-education.com/presentations/2017/... · 2017....
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Robust in vitro HBV infection systems
Lefteris Michailidis, Ph.D.
Laboratory of Virology and Infectious DiseaseCharles M. Rice LaboratoryThe Rockefeller University
December 4, 2017
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NTCP discovery
Overexpression of NTCP in hepatoma cells De novo HBV infection
NTC
PDA
PI
HepG2-NTCP (clone 7)
Yan et al. eLife (2012)Ni et al., Gastroenterology (2014)Michailidis et al. Sci Rep (2017)
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HBV infection of HepG2-NTCP cells
Michailidis et al. Sci Rep (2017)
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Need for more robust systems
High titers for infectionNo virus spread
X
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HepG2-NTCP produce infectious HBV
…but there is no spread Michailidis et al. Sci Rep (2017)
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Continuous addition of PEG results in HBV spread
Michailidis et al. Sci Rep (2017)
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Complete HBV life cycle
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HCV, YFV, WNV, VEEV, HIV-1, AdV, Flu…
Schoggins et al., Nature (2011)
Schoggins et al., Nature (2014)
Pan-viral specificity of IFN-induced genes reveals new roles for cGAS in innate immunity
A serpin shapes the extracellular environment to prevent influenza A virus maturation
Dittmann et al., Cell (2015)
ISG screens in the Rice lab
A diverse range of gene products are effectors of the type I interferon antiviral response.
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Anti-HBV ISG screens
Transduce with lenti-ISG library
Day -2 0 1
Wash/±PEG
7 13
Fix Fix
CMV ISG EMCVIRES RFP
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HBV+MyrB HBV
Anti-HBV ISG screens
DAPIDAPI
RFP
HBcAg
Total number of cells
Transduced cells
HBcAg positive cells
Non-transduced Transduced
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+ PEG
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+ PEG
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+ PEG
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ISG20
Yan et al., JVI (2015)Lv et al., Sci Rep (2015)Leong et al., Oncotarget (2016)Liu et al. PloS Pathogens (2017)Michailidis et al. Sci Rep (2017)
Control
Tetherin
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Continuous PEG treatment facilitates HBV spread in HepG2-NTCP cells
HBV spread system can be efficiently applied in antiviral screens
Summary
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Acknowledgments The Rockefeller UniversityCharles M. RiceYpe de JongJonathan PabonPaul ParkWilliam SchneiderHeinrich HoffmannAlison AshbrookXianfang WuRice lab
AlumniAmir ShlomaiTel Aviv University
Kuanhui Xiang Beijing University
CollaboratorsAmir ShlomaiTel Aviv University
Sangeeta N. BhatiaVyas RamananMIT
ReagentsHaitao GuoIndiana University
Stephan UrbanHeidelberg University
Robertson Therapeutic Development Fund
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