Hepatitis C Virus interaction with the innate immune system in HepG2-HFL cells

丙型肝炎病毒与 HepG2-HFL 细胞中先天免疫系统的相互作用

• 批准号: 8512570
• 负责人: Benjamin Goldman-Israelow
• 金额: $ 4.72万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8512570
• 关键词: Acute Liver Failure; Adverse effects; Affect; Animal Model; Antiviral Agents; Antiviral Response; Biological Models; CD81 gene; Cell Communication; Cell Culture Techniques; Cell Line; Cells; Chronic; Chronic Hepatitis C; Cirrhosis; Cleaved cell; Development; Disease; Employee Strikes; Exhibits; Genes; Goals; HCV Vaccine; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Immune; Immune response; Immune system; Impairment; Infection; Inflammation; Inflammatory; Interferons; Life; Life Cycle Stages; Link; Liver; Liver diseases; Malignant neoplasm of liver; MicroRNAs; Mitochondria; Modeling; Molecular; Molecular Biology Techniques; Nature; Outcome; Output; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Play; Population; Primary carcinoma of the liver cells; Process; Production; RNA Helicase; Reaction; Research; Role; Side; Signal Transduction; Solutions; Staging; Stimulus; System; Testing; Therapeutic; Time; Tretinoin; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; adaptive immunity; base; combat; cytokine; human TLR3 protein; improved; in vivo; insight; liver injury; liver transplantation; melanoma; novel; novel therapeutics; pathogen; prevent; public health relevance; receptor; research study; response; sensor; viral RNA; virus host interaction; virus pathogenesis

DESCRIPTION (provided by applicant): Hepatitis C virus is the leading cause of liver disease in the Western hemisphere. The current HCV therapy is inadequate, as it is associated with severe side effects and often not effective. The majority of HCV infections are persistent, life-long chronic infections that slowly induce pathogenesis. Furthermore, HCV does not appear to directly induce liver disease, rather it does so indirectly by stimulating cytokine production and inflammation, which suggests that its impairment of innate immune pathways is not absolute. The virus-host interactions and molecular mechanisms that support persistent HCV replication and chronic infection, and those that eventually induce liver disease are not understood. We hypothesize that innate intracellular immune defenses play an essential role in regulating HCV replication and that viral persistence is linked to HCV control of innate defense processes. Nearly all the studies of this topic have been conducted in Huh-7 cells or derivatives, which exhibit severely impaired innate immune responses. We recently showed that hepatoma-derived HepG2 cells efficiently support the entire HCV life cycle if the liver-specific microRNA miR-122 is expressed along with the missing entry factor CD-81 (HepG2-HFL cells). While HepG2 cells containing these two factors can replicate the entire life cycle of HCV, we found that there was a striking difference in these cells' ability to support infection and spread when compared to the previously established Huh-7.5 cell line. At the same time, we found that the innate immune response in these cells was more active in response to HCV infection. Therefore, we hypothesize that HepG2-HFL cells will faithfully recapitulate the innate immune responses present in hepatocytes, and thus provide a better model through which to understand the interactions between HCV and the innate immune system. Our ultimate goal is to better understand how HCV can subvert the antiviral response while allowing for activation of inflammatory pathways. We propose experiments to compare innate immune pathways in HepG2-HFL cells with those of primary human hepatocytes and Huh-7 cells, with a particular focus on the responses induced by the cellular viral RNA sensors RIG-I, MDA5, and TLR3. We will next examine the interaction between HCV and innate immune pathways in HepG2-HFL cells to determine whether HCV activates innate immune responses that can effectively combat infection as well as inhibit such responses through the action of viral proteins. We anticipate tha the results of these studies will provide novel insights into the interaction between HCV and the innate immune system, which remains poorly understood due to a lack of robust model systems, despite its importance in the pathogenesis of associated disease.

描述（由申请人提供）：丙型肝炎病毒是西半球肝脏疾病的主要原因。目前的HCV治疗是不够的，因为它与严重的副作用有关，而且往往无效。大多数HCV感染是持续的、终身的慢性感染，缓慢地诱导发病。此外，HCV似乎不直接诱导肝脏疾病，而是通过刺激细胞因子产生和炎症间接诱导，这表明其对先天免疫途径的损害不是绝对的。支持HCV持续复制和慢性感染的病毒-宿主相互作用和分子机制，以及最终诱导肝脏疾病的机制尚不清楚。我们假设先天性细胞内免疫防御在调节HCV复制中起重要作用，并且病毒持续存在与HCV控制先天防御过程有关。几乎所有关于这一主题的研究都是在Huh-7细胞或衍生物中进行的，这些细胞或衍生物表现出严重受损的先天免疫反应。我们最近表明，如果肝脏特异性微小RNA miR-122与缺失的进入因子CD-81一起沿着表达（HepG 2-HFL细胞），肝癌衍生的HepG 2细胞可以有效支持整个HCV生命周期。虽然含有这两种因子的HepG 2细胞可以复制HCV的整个生命周期，但我们发现，与先前建立的Huh-7.5细胞系相比，这些细胞支持感染和传播的能力存在显著差异。与此同时，我们发现这些细胞中的先天免疫反应在对HCV感染的反应中更加活跃。因此，我们假设HepG 2-HFL细胞将忠实地再现肝细胞中存在的先天免疫应答，从而提供一个更好的模型，通过该模型来了解HCV和先天免疫系统之间的相互作用。我们的最终目标是更好地了解HCV如何破坏抗病毒反应，同时允许激活炎症通路。我们提出了实验，比较先天免疫途径在HepG 2-HFL细胞与原代人肝细胞和Huh-7细胞，特别关注的细胞病毒RNA传感器RIG-I，MDA 5和TLR 3诱导的反应。接下来，我们将研究HCV和HepG 2-HFL细胞中先天免疫途径之间的相互作用，以确定HCV是否激活先天免疫应答，从而有效地对抗感染，并通过病毒蛋白的作用抑制这种应答。我们预计，这些研究的结果将提供新的见解HCV和先天免疫系统之间的相互作用，这仍然是知之甚少，由于缺乏强大的模型系统，尽管其在相关疾病的发病机制的重要性。