Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells.
Productive HBV infection of well-differentiated, hNTCP-expressing human hepatoma-derived (Huh7) cells.
复制标题
高度分化、表达 hNTCP 的人肝癌来源 (Huh7) 细胞的生产性 HBV 感染
DOI：
10.1007/s12250-017-3983-x
复制
发表时间：
2017-12
影响因子：
5.5
通讯作者：
Wu C
中科院分区：
文献类型：
--
作者：
Zhou M;Zhao K;Yao Y;Yuan Y;Pei R;Wang Y;Chen J;Hu X;Zhou Y;Chen X;Wu C
研究B型肝炎病毒（HBV）的整个生命周期，包括病毒进入的早期步骤，需要可行和有效的细胞模型。对二甲亚砜/聚乙二醇（DMSO/PEG）的抗性、hNTCP表达和分化状态是成功HBV感染模型的限制因素。在本研究中，我们使用了肝癌细胞系（Huh 7 DhNTCP），以克服这些限制因素，使其表现出良好的易感性HBV感染。为了实现这一目标，用2.5%DMSO/4%PEG 8000测试不同的肝癌细胞系，并且使用一种抗性细胞系（Huh 7 D）使用重组慢病毒系统构建稳定的hNTCP表达细胞系（Huh 7 DhNTCP）。然后，对DMSO处理前后的Huh 7 DhNTCP细胞进行形态学特征和分化分子标志物的鉴定。最后，评估Huh 7 DhNTCP细胞对HBV感染的易感性。结果表明，Huh 7 D细胞对2.5%DMSO/4%PEG 8000具有耐药性，而其他细胞则无耐药性。建立Huh 7 DhNTCP细胞以表达与肝提取物相比高水平的hNTCP，并且Huh 7 DhNTCP细胞在DMSO处理下迅速转化成非分裂的、分化良好的极化表型。Huh 7 DhNTCP细胞完全支持HBV感染的整个生命周期。这种细胞培养系统将有助于分析宿主-病毒相互作用，这将有助于发现抗病毒药物和疫苗。可通过10.1007/s12250-017-3983-x获取本文的补充材料，授权用户可访问。
Feasible and effective cell models for hepatitis B virus (HBV) infection are required for investigating the complete lifecycle of this virus, including the early steps of viral entry. Resistance to dimethyl sulfoxide/polyethylene glycol (DMSO/PEG), hNTCP expression, and a differentiated state are the limiting factors for successful HBV infection models. In the present study, we used a hepatoma cell line (Huh7DhNTCP) to overcome these limiting factors so that it exhibits excellent susceptibility to HBV infection. To achieve this goal, different hepatoma cell lines were tested with 2.5% DMSO / 4% PEG8000, and one resistant cell line (Huh7D) was used to construct a stable hNTCP-expressing cell line (Huh7DhNTCP) using a recombinant lentivirus system. Then, the morphological characteristics and differentiation molecular markers of Huh7DhNTCP cells with or without DMSO treatment were characterized. Finally, the susceptibility of Huh7DhNTCP cells to HBV infection was assessed. Our results showed that Huh7D cells were resistant to 2.5% DMSO / 4% PEG8000, whereas the others were not. Huh7DhNTCP cells were established to express a high level of hNTCP compared to liver extracts, and Huh7DhNTCP cells rapidly transformed into a non-dividing, well-differentiated polarized phenotype under DMSO treatment. Huh7DhNTCP cells fully supported the entire lifecycle of HBV infection. This cell culture system will be useful for the analysis of host-virus interactions, which should facilitate the discovery of antiviral drugs and vaccines. Supplementary material is available for this article at 10.1007/s12250-017-3983-x and is accessible for authorized users.
登录
查看更多内容
影响因子：
5.4
作者：
Köck, J;Nassal, M;von Weizsäcker, F
通讯作者：
von Weizsäcker, F
影响因子：
25.7
作者：
Shimura S;Watashi K;Fukano K;Peel M;Sluder A;Kawai F;Iwamoto M;Tsukuda S;Takeuchi JS;Miyake T;Sugiyama M;Ogasawara Y;Park SY;Tanaka Y;Kusuhara H;Mizokami M;Sureau C;Wakita T
通讯作者：
Wakita T
影响因子：
5.4
作者：
GRIPON, P;DIOT, C;GUGUENGUILLOUZO, C
通讯作者：
GUGUENGUILLOUZO, C
影响因子：
13.5
作者：
Kawai, HF;Kaneko, S;Kobayashi, K
通讯作者：
Kobayashi, K
DOI：
10.1007/978-1-4939-6700-1_2
发表时间：
2017-01-01
期刊：
HEPATITIS B VIRUS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ni, Yi;Urban, Stephan
通讯作者：
Urban, Stephan