Entry and Release of Hepatitis C Virus in Polarized Human Hepatocytes
Entry and Release of Hepatitis C Virus in Polarized Human Hepatocytes
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DOI：
10.1128/jvi.00478-17
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发表时间：
2017-09-01
影响因子：
5.4
通讯作者：
Dubuisson, Jean
中科院分区：
文献类型：
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作者：
Belouzard, Sandrine;Danneels, Adeline;Dubuisson, Jean
丙型肝炎病毒（HCV）主要感染肝细胞，肝细胞是高度极化的细胞。细胞极性在HCV生命周期中的相关性仅在远亲模型中得到解决，并且仍然知之甚少。虽然极化上皮细胞有一个相当简单的形态与基底外侧和顶端域，肝细胞表现出复杂的极化结构。然而，据报道，一些选定的极化HepG 2细胞克隆可以表现出典型的柱状极化上皮细胞的紧密连接蛋白的蜂窝状分布模式，这可以用作一个简单的模型来研究细胞极化在病毒感染肝细胞中的作用。为了获得类似的克隆，使用表达CD 81的HepG 2细胞（HepG 2-CD 81），并通过有限稀释分离克隆。两个克隆表现出一个简单的柱状极化能力时，生长在半渗透性支持分离和表征。为了测试HCV进入和释放的极性，用细胞培养物衍生的HCV感染我们的极化HepG 2-CD 81克隆。我们的数据表明，HCV与细胞两侧的结合量相同，但主要当病毒添加到基底外侧区时，会发生有效感染。此外，我们还观察到HCV病毒体从细胞的基底外侧域释放。最后，当用油酸和MEK抑制剂U 0126处理极化细胞以促进脂蛋白分泌时，分泌出更高比例的低密度感染性病毒颗粒。这种细胞培养系统为研究细胞极化对HCV生命周期的影响提供了一个极好的模型。重要信息丙型肝炎是一种主要的健康负担，全世界约有1.7亿人感染。丙型肝炎病毒（HCV）主要感染肝细胞，肝细胞是具有复杂组织的高度极化细胞。细胞极性在HCV生命周期中的相关性已经在远亲模型中得到了解决，但仍不清楚。肝细胞组织结构复杂，具有多个顶侧和基底侧表面。开发了一种表达CD 81的HepG 2细胞的简单培养模型来研究HCV感染，所述细胞能够与独特的顶侧和基底侧结构域结合。利用该模型，我们证明了HCV通过基底外侧域进入和离开肝细胞。此外，在促进脂蛋白分泌的条件下产生低密度病毒颗粒。这种细胞培养系统为研究细胞极化对HCV感染的影响提供了一个有用的模型。
Hepatitis C virus (HCV) primarily infects hepatocytes, which are highly polarized cells. The relevance of cell polarity in the HCV life cycle has been addressed only in distantly related models and remains poorly understood. Although polarized epithelial cells have a rather simple morphology with a basolateral and an apical domain, hepatocytes exhibit complex polarization structures. However, it has been reported that some selected polarized HepG2 cell clones can exhibit a honeycomb pattern of distribution of the tight-junction proteins typical of columnar polarized epithelia, which can be used as a simple model to study the role of cell polarization in viral infection of hepatocytes. To obtain similar clones, HepG2 cells expressing CD81 (HepG2-CD81) were used, and clones were isolated by limiting dilutions. Two clones exhibiting a simple columnar polarization capacity when grown on a semipermeable support were isolated and characterized. To test the polarity of HCV entry and release, our polarized HepG2-CD81 clones were infected with cell culture-derived HCV. Our data indicate that HCV binds equally to both sides of the cells, but productive infection occurs mainly when the virus is added at the basolateral domain. Furthermore, we also observed that HCV virions are released from the basolateral domain of the cells. Finally, when polarized cells were treated with oleic acid and U0126, a MEK inhibitor, to promote lipoprotein secretion, a higher proportion of infectious viral particles of lower density were secreted. This cell culture system provides an excellent model to investigate the influence of cell polarization on the HCV life cycle.IMPORTANCE Hepatitis C is a major health burden, with approximately 170 million persons infected worldwide. Hepatitis C virus (HCV) primarily infects hepatocytes, which are highly polarized cells with a complex organization. The relevance of cell polarity in the HCV life cycle has been addressed in distantly related models and remains unclear. Hepatocyte organization is complex, with multiple apical and basolateral surfaces. A simple culture model of HepG2 cells expressing CD81 that are able to polarize with unique apical and basolateral domains was developed to study HCV infection. With this model, we demonstrated that HCV enters and exits hepatocytes by the basolateral domain. Furthermore, lower-density viral particles were produced under conditions that promote lipoprotein secretion. This cell culture system provides a useful model to study the influence of cell polarization on HCV infection.