Intracellular delivery of serum-derived hepatitis C virus.
A robust and reliable cell culture system for serum-derived HCV (HCVser) has not been established yet because of the presence of neutralizing antibody and tropism for infection. To overcome this obstacle, we employed a lipid-mediated protein intracellular delivery reagent (PIDR) that permits internalization of proteins into cells. Although entry of HCVcc was not enhanced by the treatment with PIDR, entry of HCVser into hepatoma cell lines (Huh7 and HepG2) and immortalized primary hepatocytes (Hc and HuS/E2) was significantly enhanced by the PIDR treatment. The entry of HCVser into Huh7 cells in the presence of PIDR was resistant to the neutralization by an anti-hCD81 antibody, suggesting that PIDR is capable of internalizing HCVser in a receptor-independent manner. Interestingly, the PIDR-mediated entry of HCVser and HCVcc was enhanced by the addition of sera from chronic hepatitis C patients but not from healthy donors. In addition, neutralization of HCVcc infection by anti-E2 antibody was canceled by the treatment with PIDR. In conclusion, the PIDR is a valuable tool to get over the obstacle of neutralizing antibodies to internalize HCV into cells and might be useful for the establishment of in vitro propagation HCVser.
Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.
Takasuke
T
Fukuhara
Hideki
H
Tani
Mai
M
Shiokawa
Yukinori
Y
Goto
Takayuki
T
Abe
Akinobu
A
Taketomi
Ken
K
Shirabe
Yoshihiko
Y
Maehara
Yoshiharu
Y
Matsuura
D057134:Antibodies, Neutralizing / Q000097:blood / Q000378:metabolism; D000914:Antibodies, Viral / Q000097:blood / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D002460:Cell Line; D057809:HEK293 Cells; D056945:Hep G2 Cells; D016174:Hepacivirus / Q000276:immunology / Q000502:physiology*; D006494:Heparin Antagonists / Q000494:pharmacology; D019698:Hepatitis C, Chronic / Q000276:immunology*; D022781:Hepatocytes / Q000187:drug effects / Q000473:pathology / Q000821:virology; D006583:Hexadimethrine Bromide / Q000494:pharmacology; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism / Q000821:virology*; D021381:Protein Transport; D044967:Serum / Q000821:virology*; D060225:Tetraspanin 28; D053586:Virus Internalization / Q000187:drug effects
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D015703:Antigens, CD; C554798:CD81 protein, human; D006494:Heparin Antagonists; D060225:Tetraspanin 28; D006583:Hexadimethrine Bromide
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