Construction of the Vero cell culture system that can produce infectious HCV particles.
The hepatitis C virus is a major cause of chronic liver disease worldwide. Lack of culture system supporting virus production has been one of the major impediments in HCV research and vaccine development. Here, we use a HCV (1b) full-length cDNA clone that replicates and produces integrated and infectious virus particles in cultured Vero cells. Evidence shows that the replication of virus particles is robust, producing over 10(8) copies of positive RNA per milliliter of the culture cells within 48 h. Sucrose density gradient centrifugation of the cell lysate reveals that the HCV virions have a density of about 1.17 g/ml and a spherical morphology with an average diameter of about 55 nm. Secreted virus is infectious for Huh7 cells and can be neutralized by CD81- and E2-specific antibodies. This system establishes a powerful framework for studying the virus life cycle and developing vaccine research.
Guo
Jia
J
College of Life Sciences, Wuhan University, Luojia Mountain, Wuhan, 430072, China.
Yan
Ran
R
Xu
Guodong
G
Li
Weiyun
W
Zheng
Congyi
C
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015870:Gene Expression; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000378:metabolism; D060225:Tetraspanin 28; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D014771:Virion / Q000502:physiology* / Q000648:ultrastructure; D014776:Virus Cultivation / Q000379:methods*
D000906:Antibodies; D015703:Antigens, CD; C554798:CD81 protein, human; D012367:RNA, Viral; D060225:Tetraspanin 28; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C091015:glycoprotein E2, Hepatitis C virus