Intracellular delivery of serum-derived hepatitis C virus.
Intracellular delivery of serum-derived hepatitis C virus.
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DOI：
10.1016/j.micinf.2011.01.005
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发表时间：
2011-04
影响因子：
5.8
通讯作者：
Matsuura Y
中科院分区：
文献类型：
--
作者：
Fukuhara T;Tani H;Shiokawa M;Goto Y;Abe T;Taketomi A;Shirabe K;Maehara Y;Matsuura Y
由于血清源性HCV（HCVser）存在中和抗体和感染嗜性，尚未建立稳定可靠的细胞培养系统。为了克服这一障碍，我们采用了脂质介导的蛋白质细胞内递送试剂（PIDR），允许蛋白质内化到细胞中。尽管用PIDR处理没有增强HCVser的进入，但PIDR处理显著增强HCVser进入肝癌细胞系（Huh 7和HepG 2）和永生化原代肝细胞（Hc和HuS/E2）。在PIDR存在下，HCVser进入Huh 7细胞对抗hCD 81抗体的中和具有抗性，表明PIDR能够以受体非依赖性方式内化HCVser。有趣的是，PIDR介导的HCVser和HCvser的进入通过添加来自慢性丙型肝炎患者的血清而不是来自健康供体的血清而增强。此外，抗E2抗体对HCV-infection的中和作用被PIDR治疗所取消。因此，PIDR是克服中和抗体将HCV内化到细胞中的障碍的有价值的工具，并且可能用于体外增殖HCVser的建立。
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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