Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells, and Primary Human Hepatocytes
Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells, and Primary Human Hepatocytes
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DOI：
10.1007/978-1-4939-6700-1_2
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发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Urban, Stephan
中科院分区：
文献类型：
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作者：
Ni, Yi;Urban, Stephan
病毒-宿主相互作用的研究依赖于有效支持病毒感染的合适的体外细胞培养系统。这样的系统应该理想地提供类似于天然宿主细胞的条件，例如，细胞类型特异性信号传导和代谢途径。对于HBV感染，原代人肝细胞（PHH）是满足这些要求的最可靠的系统，但获得这些细胞是有限的。此外，实验结果的再现性取决于许多因素，包括制备方法或供体的可变性。分化后的人肝细胞系HepaRG在许多肝细胞特异性标志物方面类似于PHH，包括细胞色素P450酶、肝特异性转录因子和转运蛋白如HBV特异性受体、牛磺胆酸钠共转运多肽（NTCP）的表达。HepaRG细胞也被证明表达先天免疫系统的关键分子。到目前为止，HepaRG细胞系是唯一一个允许研究HBV/HDV感染和肝脏特异性药物毒性和代谢的细胞系。与PHH相比，HepaRG细胞相对较低的易感性取决于各种因素，并且可以通过受体NTCP的组成型表达部分克服，从而允许感染而不完全分化。异位NTCP表达不影响DMSO诱导的细胞分化能力。在这里，我们详细描述了如何在技术上进行HBV感染体外与这些细胞。该方法可用于探索HBV感染的机制，并建立适合于在代谢上接近原代人肝细胞的细胞中评价药物疗效的抗病毒筛选平台。
Investigations of virus-host interactions rely on suitable in vitro cell culture systems that efficiently support virus infection. Such systems should ideally provide conditions that resemble those of natural host cells, e.g., the cell-type specific signaling and metabolic pathways. For HBV infection, primary human hepatocytes (PHHs) are the most faithful system fulfilling these requirements but access to these cells is limited. Moreover, the reproducibility of experimental results depends on many factors including the preparation method or variability of the donors. The human liver cell line HepaRG, after differentiation, resembles PHHs with respect to many hepatocyte-specific markers including the expression of cytochrome P450 enzymes, liver-specific transcription factors, and transporter proteins like the HBV-specific receptor, sodium taurocholate co-transporting polypeptide (NTCP). HepaRG cells have also been shown to express key molecules of the innate immune system. So far, the HepaRG cell line is the only one allowing both studies on HBV/HDV infection and liver-specific drug toxicity and metabolism. The relative low susceptibility of HepaRG cells when compared with PHHs depends on various factors and can partially be overcome by constitutive expression of the receptor NTCP, allowing infection without full differentiation. Ectopic NTCP expression does not interfere with the ability of cell differentiation induced by DMSO. Here, we describe in detail how to technically perform HBV infection in vitro with these cells. The methods can be used to explore the mechanism of HBV infection and to build an antiviral screening platform suitable for evaluation of drug efficacy in cells that are metabolically close to primary human hepatocytes.