Department of Infectious Diseases, Molecular Virology, Heidelberg University, Heidelberg, Germany; Division of Virus-Associated Carcinogenesis (F170), German Cancer Research Center (DKFZ), Heidelberg, Germany; HBIGS graduate school, Heidelberg, Germany.
Department of Infectious Diseases, Molecular Virology, Heidelberg University, Heidelberg, Germany.
Gastroenterology. 2018 May;154(6):1791-1804.e22. doi: 10.1053/j.gastro.2018.01.044. Epub 2018 Feb 1.
BACKGROUND & AIMS: Hepatitis C virus (HCV) infection is sensitive to interferon (IFN)-based therapy, whereas hepatitis B virus (HBV) infection is not. It is unclear whether HBV escapes detection by the IFN-mediated immune response or actively suppresses it. Moreover, little is known on how HBV and HCV influence each other in coinfected cells. We investigated interactions between HBV and the IFN-mediated immune response using HepaRG cells and primary human hepatocytes (PHHs). We analyzed the effects of HBV on HCV replication, and vice versa, at the single-cell level.
PHHs were isolated from liver resection tissues from HBV-, HCV-, and human immunodeficiency virus-negative patients. Differentiated HepaRG cells overexpressing the HBV receptor sodium taurocholate cotransporting polypeptide (dHepaRGNTCP) and PHHs were infected with HBV. Huh7.5 cells were transfected with circular HBV DNA genomes resembling viral covalently closed circular DNA (cccDNA), and subsequently infected with HCV; this served as a model of HBV and HCV coinfection. Cells were incubated with IFN inducers, or IFNs, and antiviral response and viral replication were analyzed by immune fluorescence, reverse-transcription quantitative polymerase chain reaction, enzyme-linked immunosorbent assays, and flow cytometry.
HBV infection of dHepaRGNTCP cells and PHHs neither activated nor inhibited signaling via pattern recognition receptors. Incubation of dHepaRGNTCP cells and PHHs with IFN had little effect on HBV replication or levels of cccDNA. HBV infection of these cells did not inhibit JAK-STAT signaling or up-regulation of IFN-stimulated genes. In coinfected cells, HBV did not prevent IFN-induced suppression of HCV replication.
In dHepaRGNTCP cells and PHHs, HBV evades the induction of IFN and IFN-induced antiviral effects. HBV infection does not rescue HCV from the IFN-mediated response.
丙型肝炎病毒（HCV）感染对干扰素（IFN）为基础的治疗敏感，而乙型肝炎病毒（HBV）感染则不然。尚不清楚 HBV 是否通过 IFN 介导的免疫反应逃避检测，还是主动抑制它。此外，对于 HBV 和 HCV 在共感染细胞中如何相互影响知之甚少。我们使用 HepaRG 细胞和原代人肝细胞（PHH）研究了 HBV 与 IFN 介导的免疫反应之间的相互作用。我们分析了 HBV 对 HCV 复制的影响，反之亦然，在单细胞水平上。
从 HBV、HCV 和人类免疫缺陷病毒阴性患者的肝切除组织中分离 PHH。表达 HBV 受体牛磺胆酸钠共转运蛋白（dHepaRGNTCP）的分化 HepaRG 细胞和 PHH 被 HBV 感染。Huh7.5 细胞用类似于病毒共价闭合环状 DNA（cccDNA）的环形 HBV DNA 基因组转染，并随后感染 HCV；这作为 HBV 和 HCV 共感染的模型。用 IFN 诱导剂或 IFN 孵育细胞，并通过免疫荧光、逆转录定量聚合酶链反应、酶联免疫吸附测定和流式细胞术分析抗病毒反应和病毒复制。
HBV 感染 dHepaRGNTCP 细胞和 PHH 既没有激活也没有抑制模式识别受体的信号转导。IFN 孵育对 dHepaRGNTCP 细胞和 PHH 的 HBV 复制或 cccDNA 水平几乎没有影响。HBV 感染这些细胞不会抑制 JAK-STAT 信号转导或 IFN 刺激基因的上调。在共感染细胞中，HBV 不能阻止 IFN 诱导的 HCV 复制抑制。
在 dHepaRGNTCP 细胞和 PHH 中，HBV 逃避 IFN 的诱导和 IFN 诱导的抗病毒作用。HBV 感染不能使 HCV 免受 IFN 介导的反应。
