Expression of paramyxovirus V proteins promotes replication and spread of hepatitis C virus in cultures of primary human fetal liver cells.
Expression of paramyxovirus V proteins promotes replication and spread of hepatitis C virus in cultures of primary human fetal liver cells.
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DOI：
10.1002/hep.24557
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发表时间：
2011-12
期刊：
影响因子：
13.5
通讯作者：
Rice, Charles M.
中科院分区：
文献类型：
--
作者：
Andrus, Linda;Marukian, Svetlana;Jones, Christopher T.;Catanese, Maria Teresa;Sheahan, Timothy P.;Schoggins, John W.;Barry, Walter T.;Dustin, Lynn B.;Trehan, Kartik;Ploss, Alexander;Bhatia, Sangeeta N.;Rice, Charles M.
在这里，我们证明了人胎肝细胞(Hflc)的原代培养可靠地支持丙型肝炎病毒(HCV)实验室毒株的感染，尽管不同供体细胞制剂之间的病毒复制水平显著不同，并且经常以一种提示病毒主动清除的方式下降。为了研究干扰素系统对控制人肝细胞癌中丙型肝炎病毒感染的可能作用，我们利用副粘病毒(PMV)V蛋白的特性来对抗干扰素诱导和抗病毒信号。用慢病毒载体将麻疹病毒(MV)和副流感病毒5(PIV5)的V蛋白导入hflc，以显示丙型肝炎病毒感染的细胞。V蛋白转导的hflc支持与未转导或对照载体转导的hflc相比，丙型肝炎病毒感染水平增加(10至100倍)。通过病毒驱动的荧光素酶的测定、感染性丙型肝炎病毒和病毒RNA的分析以及感染丙型肝炎病毒的肝细胞的直接观察来评估感染情况。在感染后48至119小时之间的活细胞成像显示，在没有PMV V蛋白表达的情况下，感染很少或没有扩散。相比之下，V蛋白转导的hflc显示了许多丙型肝炎病毒感染事件。V蛋白的表达可有效拮抗加入干扰素的人肝癌细胞中丙型肝炎病毒的抑制作用。此外，在V蛋白转导的培养物中，急性丙型肝炎病毒感染后诱导的III型干扰素IL29被抑制。结论：这些研究表明，细胞内的干扰素应答在限制原代培养的肝细胞中丙型肝炎病毒感染的传播中起重要作用。旨在抑制这种反应的策略可能是进一步发展强大的丙型肝炎病毒培养系统的关键，从而能够研究病毒的致病性和丙型肝炎病毒在其自然宿主细胞群体中传播的机制。(《肝病》2011；54：1901-1912)
Here we demonstrate that primary cultures of human fetal liver cells (HFLC) reliably support infection with laboratory strains of hepatitis C virus (HCV), although levels of virus replication vary significantly between different donor cell preparations and frequently decline in a manner suggestive of active viral clearance. To investigate possible contributions of the interferon (IFN) system to control HCV infection in HFLC, we exploited the well-characterized ability of paramyxovirus (PMV) V proteins to counteract both IFN induction and antiviral signaling. The V proteins of measles virus (MV) and parainfluenza virus 5 (PIV5) were introduced into HFLC using lentiviral vectors encoding a fluorescent reporter for visualization of HCV-infected cells. V protein-transduced HFLC supported enhanced (10 to 100-fold) levels of HCV infection relative to untransduced or control vector-transduced HFLC. Infection was assessed by measurement of virus-driven luciferase, by assays for infectious HCV and viral RNA, and by direct visualization of HCV-infected hepatocytes. Live cell imaging between 48 and 119 hours postinfection demonstrated little or no spread of infection in the absence of PMV V protein expression. In contrast, V protein-transduced HFLC showed numerous HCV infection events. V protein expression efficiently antagonized the HCV-inhibitory effects of added IFNs in HFLC. In addition, induction of the type III IFN, IL29, following acute HCV infection was inhibited in V protein-transduced cultures. Conclusion: These studies suggest that the cellular IFN response plays a significant role in limiting the spread of HCV infection in primary hepatocyte cultures. Strategies aimed at dampening this response may be key to further development of robust HCV culture systems, enabling studies of virus pathogenicity and the mechanisms by which HCV spreads in its natural host cell population. (Hepatology 2011;54:1901-1912)
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影响因子：
5.4
作者：
Kohaar, Indu;Ploss, Alexander;Prokunina-Olsson, Ludmila
通讯作者：
Prokunina-Olsson, Ludmila
影响因子：
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Horvath, CA
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Horvath, Curt M.
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13.5
作者：
Banaudha, Krishna;Orenstein, Jan M.;Kumar, Ajit
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Kumar, Ajit