Letter | Published:

SEC14L2 enables pan-genotype HCV replication in cell culture

Nature volume 524, pages 471475 (27 August 2015) | Download Citation


Since its discovery in 1989, efforts to grow clinical isolates of the hepatitis C virus (HCV) in cell culture have met with limited success. Only the JFH-1 isolate has the capacity to replicate efficiently in cultured hepatoma cells without cell culture-adaptive mutations1,2,3. We hypothesized that cultured cells lack one or more factors required for the replication of clinical isolates. To identify the missing factors, we transduced Huh-7.5 human hepatoma cells with a pooled lentivirus-based human complementary DNA (cDNA) library, transfected the cells with HCV subgenomic replicons lacking adaptive mutations, and selected for stable replicon colonies. This led to the identification of a single cDNA, SEC14L2, that enabled RNA replication of diverse HCV genotypes in several hepatoma cell lines. This effect was dose-dependent, and required the continuous presence of SEC14L2. Full-length HCV genomes also replicated and produced low levels of infectious virus. Remarkably, SEC14L2-expressing Huh-7.5 cells also supported HCV replication following inoculation with patient sera. Mechanistic studies suggest that SEC14L2 promotes HCV infection by enhancing vitamin E-mediated protection against lipid peroxidation. This provides a foundation for development of in vitro replication systems for all HCV isolates, creating a useful platform to dissect the mechanisms by which cell culture-adaptive mutations act.

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We thank Y. Matsuura for HEP3B/miR-122 cells, J. Bukh for pJ6CF and pTNcc/GLuc, S. M. Lemon and D. Yamane for pH77S.3/GLuc, pH77S.3, and pH77D/GLuc, and D. Manor for [14C]RRR-α-tocopherol. We also thank E. Castillo and A. Webson for laboratory assistance, H. H. Hoffman and L. Andrus for technical input, and W. M. Schneider, M. M. Li and M. R. MacDonald for critical reading of the manuscript. This work was supported in part by the National Institutes of Health, NCI grant R01CA057973, NIAID grants R01AI072613 and R01AI099284 (to C.M.R.), NIDDK grant R01DK090317 and NIDA grant DA031095 (to A.D.B.), by a Helmsley Postdoctoral Fellowship for Basic and Translational Research on Disorders of the Digestive System at The Rockefeller University (to M.S.), and by a Liver Scholar Award from American Association for the Study of Liver Diseases (U.A.). The Greenberg Medical Research Institute, the Starr Foundation, the Ronald A. Shellow, M.D. Memorial Fund and anonymous donors provided the additional funding (to C.M.R.).

Author information


  1. Center for the Study of Hepatitis C, Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York 10065, USA

    • Mohsan Saeed
    • , Ursula Andreo
    • , Hyo-Young Chung
    • , Christine Espiritu
    •  & Charles M. Rice
  2. Division of Liver Diseases, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA

    • Andrea D. Branch
  3. Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA

    • Jose M. Silva


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M.S. and C.M.R. designed the project, analysed results, and wrote the manuscript. M.S., U.A., H.-Y.C. and C.E. performed the experimental work. A.D.B. and J.M.S. contributed reagents and advice.

Competing interests

C.M.R. has equity in Apath, LLC, which holds commercial licenses for the Huh-7.5 cell line and certain HCV cell culture systems.

Corresponding author

Correspondence to Charles M. Rice.

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