Journal
MICROBES AND INFECTION
Volume 13, Issue 4, Pages 405-412Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.micinf.2011.01.005
Keywords
Hepatitis C virus; Protein intracellular delivery; Serum-derived virus
Categories
Funding
- Ministry of Health, Labor, and Welfare
- Ministry of Education, Culture, Sports, Science, and Technology, Japan
- Grants-in-Aid for Scientific Research [21390138, 22659246, 21249072, 23790502] Funding Source: KAKEN
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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 (He 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. (C) 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.
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