Production of infectious hepatitis C virus particles in three-dimensional cultures of the cell line carrying the genome-length dicistronic viral RNA of genotype 1b.
Production of infectious hepatitis C virus particles in three-dimensional cultures of the cell line carrying the genome-length dicistronic viral RNA of genotype 1b.
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DOI：
10.1016/j.virol.2006.03.038
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发表时间：
2006-08
期刊：
影响因子：
3.7
通讯作者：
Kyoko Murakami;K. Ishii;Y. Ishihara;Sayaka Yoshizaki;Keiko Tanaka;Y. Gotoh;H. Aizaki;M. Kohara;H. Yoshioka;Y. Mori;N. Manabe;I. Shoji;T. Sata;R. Bartenschlager;Y. Matsuura;T. Miyamura;Tetsuro Suzuki
中科院分区：
文献类型：
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作者：
Kyoko Murakami;K. Ishii;Y. Ishihara;Sayaka Yoshizaki;Keiko Tanaka;Y. Gotoh;H. Aizaki;M. Kohara;H. Yoshioka;Y. Mori;N. Manabe;I. Shoji;T. Sata;R. Bartenschlager;Y. Matsuura;T. Miyamura;Tetsuro Suzuki
我们发现，双顺反子丙型肝炎病毒（HCV）基因组的基因型1b支持生产和分泌的传染性HCV颗粒在两个独立的三维（3D）培养系统，径向流生物反应器和热可逆凝胶聚合物（TGP），但不是在单层培养。在Huh-7细胞球体中的培养基以及内质网膜和扩张的细胞质池中观察到直径为50-60 nm并被膜样结构包围的免疫反应性包膜颗粒。HCV颗粒的感染被干扰E2与人细胞结合的抗E2抗体或患者血清中和。最后，显示了3D-TGP培养系统用于抗病毒药物评价的实用性。我们得出结论，基于复制子的3D培养系统允许生产感染性HCV颗粒。该系统是一个有价值的工具，在自然宿主细胞环境中的HCV形态发生的研究。
We show that a dicistronic hepatitis C virus (HCV) genome of genotype 1b supports the production and secretion of infectious HCV particles in two independent three-dimensional (3D) culture systems, the radial-flow bioreactor and the thermoreversible gelation polymer (TGP), but not in monolayer cultures. Immunoreactive enveloped particles, which are 50–60 nm in diameter and are surrounded by membrane-like structures, are observed in the culture medium as well as at the endoplasmic reticulum membranes and in dilated cytoplasmic cisternae in spheroids of Huh-7 cells. Infection of HCV particles is neutralized by anti-E2 antibody or patient sera that interfere with E2 binding to human cells. Finally, the utility of the 3D-TGP culture system for the evaluation of antiviral drugs is shown. We conclude that the replicon-based 3D culture system allows the production of infectious HCV particles. This system is a valuable tool in studies of HCV morphogenesis in a natural host cell environment.