TY - JOUR
T1 - Intracellular delivery of serum-derived hepatitis C virus
AU - Fukuhara, Takasuke
AU - Tani, Hideki
AU - Shiokawa, Mai
AU - Goto, Yukinori
AU - Abe, Takayuki
AU - Taketomi, Akinobu
AU - Shirabe, Ken
AU - Maehara, Yoshihiko
AU - Matsuura, Yoshiharu
N1 - Funding Information:
This research was supported in part by grants-in-aid from the Ministry of Health, Labor, and Welfare , and the Ministry of Education, Culture, Sports, Science, and Technology, Japan .
PY - 2011/4
Y1 - 2011/4
N2 - 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.
AB - 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.
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U2 - 10.1016/j.micinf.2011.01.005
DO - 10.1016/j.micinf.2011.01.005
M3 - Article
C2 - 21262370
AN - SCOPUS:79952453062
SN - 1286-4579
VL - 13
SP - 405
EP - 412
JO - Microbes and Infection
JF - Microbes and Infection
IS - 4
ER -
