Persistence of HCV in Quiescent Hepatic Cells Under Conditions of an Interferon-Induced Antiviral Response
Persistence of HCV in Quiescent Hepatic Cells Under Conditions of an Interferon-Induced Antiviral Response
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DOI：
10.1053/j.gastro.2012.04.018
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发表时间：
2012-08-01
期刊：
影响因子：
29.4
通讯作者：
Bartenschlager, Ralf
中科院分区：
文献类型：
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作者：
Bauhofer, Oliver;Ruggieri, Alessia;Bartenschlager, Ralf
背景与目的：丙型肝炎病毒（HCV）是慢性肝病的常见原因。很多患者并没有清除病毒感染；对于 HCV 持续存在的机制以及干扰素 (IFN) 经常无法消除 HCV 的机制，人们知之甚少。需要更好的培养系统来研究静止肝细胞中的病毒复制。方法：我们使用人肝癌细胞 (Huh7.5) 和经历增殖停滞和分化的细胞 (Huh7.5(dif)) 来研究细胞暴露于 IFN-α 后 HCV 感染的持续性，并比较 IFN-α 和 IFN-lambda 的抗病毒作用。我们用表达 IFN 诱导荧光团的原代人肝细胞和 Huh7 细胞验证了这些结果。结果：感染Huh7.5(dif)细胞后，HCV持续复制并释放感染性颗粒。细胞长期暴露于 IFN-α 下，HCV 复制减少了近 1000 倍，但并没有消除病毒；停用干扰素后，病毒复制反弹，就像慢性丙型肝炎病毒感染患者的情况一样。 HCV 在对 IFN-α 反应水平较低的细胞中以较高水平复制，但并非完全如此。将细胞与等浓度的 IFN-α 或 IFN-lambda 一起孵育后，Huh7.5(dif) 细胞比未分化的 Huh7.5 细胞或原代人肝细胞表达更广泛的 IFN 刺激基因模式，表明抗病毒反应取决于细胞的分化状态。结论：我们开发了一种使用肝癌细胞的细胞培养系统，以研究 I 型或 III 型 IFN 诱导的抗病毒反应期间的持续 HCV 感染。抗病毒反应的水平和范围与细胞的分化状态相关。我们认为 HCV 利用肝细胞对 IFN 反应的随机性来维持持久性。
BACKGROUND & AIMS: Hepatitis C virus (HCV) is a common cause of chronic liver disease. Many patients do not clear the viral infection; little is known about the mechanisms of HCV persistence or the frequent failure of interferon (IFN) to eliminate it. Better culture systems are needed to study viral replication in quiescent liver cells. METHODS: We used human hepatoma (Huh7.5) cells and those that had undergone proliferation arrest and differentiation (Huh7.5(dif)) to study the persistence of HCV infection following exposure of the cells to IFN-alpha and to compare the antiviral effects of IFN-alpha and IFN-lambda. We validated these results with primary human hepatocytes and Huh7 cells that expressed an IFN-inducible fluorophore. RESULTS: Following infection of Huh7.5(dif) cells, HCV replicated persistently and released infectious particles. Long-term exposure of the cells to IFN-alpha reduced HCV replication similar to 1000-fold but did not eliminate the virus; viral replication rebounded after withdrawal of IFN, as it does in patients with chronic HCV infection. HCV replicated at higher levels, but not exclusively, in cells that had a low level of response to IFN-alpha. Following incubation of cells with equipotent concentrations of IFN-alpha or IFN-lambda, Huh7.5(dif) cells expressed a wider pattern of IFN-stimulated genes than undifferentiated Huh7.5 cells or primary human hepatocytes, indicating that the antiviral response depends on the differentiation status of the cells. CONCLUSIONS: We developed a cell culture system using hepatoma cells to study persistent HCV infection during the type I or type III IFN-induced antiviral response. The level and range of the antiviral responses were associated with the differentiation status of the cells. We propose that HCV exploits the stochastic nature of the response of hepatocytes to IFN to sustain persistence.