Damage-associated Alarmin Key to Immune Responses in Viral Hepatitis

损伤相关警报蛋白是病毒性肝炎免疫反应的关键

• 批准号: 8771220
• 负责人: JIAREN SUN
• 金额: $ 23.23万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8771220
• 关键词: Acute; Acute Hepatitis; Adenoviruses; Animal Model; Animals; Antiviral Agents; Antiviral Therapy; Binding; Biological Assay; CD8B1 gene; Cell Communication; Cell physiology; Characteristics; Chemicals; Chronic; Chronic Hepatitis; Chronic Hepatitis B; Clinical; DNA; Development; Disease; Disease Outcome; Dose; Effector Cell; Event; Flow Cytometry; Future; Goals; HMGB Proteins; Hepatic; Hepatitis; Hepatitis C; Hepatitis Viruses; Hepatitis, Chronic, Drug-Induced; Hepatotoxicity; IL7R gene; Immune; Immune System Diseases; Immune response; Immune system; Inflammation; Inflammatory; Injection of therapeutic agent; Interferon Type II; Interferons; Interleukin-13; Interleukin-5; Ischemia; Lead; Liver; Liver Failure; Liver diseases; Lymphocytic choriomeningitis virus; Lymphoid Cell; Malignant Neoplasms; Measures; Mediating; Modeling; Molecular; Mus; NF-kappa B; Nature; Nucleoproteins; Outcome; Pathogenesis; Pathway interactions; Patients; Pattern; Phenotype; Prevention; Process; Production; Property; Proto-Oncogene Protein c-kit; Recombinants; Reperfusion Therapy; Reporting; Research; Resolution; Role; Serum; Signal Pathway; Signal Transduction; Source; System; T cell response; T-Lymphocyte; TNF gene; Testing; Therapeutic; Time; Tissues; Transgenic Mice; Viral; Viral hepatitis; Virus; Virus Diseases; Work; chromatin immunoprecipitation; cytokine; human subject; improved; innovation; insight; intrahepatic; knockout animal; liver inflammation; liver injury; novel; outcome forecast; public health relevance; receptor; reconstitution; response

DESCRIPTION (provided by applicant): The critical molecular events that regulate immune control vs. disease pathogenesis in viral hepatitis remain elusive. However, recent reports suggested that patients with chronic hepatitis displayed elevated concentrations of alarmin (IL-33) and its decoy receptor sST2 in the serum. Their levels were associated with hepatic damage, viral and clinical outcomes in response to antiviral therapy. We hypothesize that IL-33 can directly engage innate and adaptive immune systems in the liver and also induce differentiation of novel group 2 innate lymphoid cells (ILC2). Through these actions, IL-33 can modulate immune responses and protect the liver during the inflammatory process. Two complementary approaches are proposed to test these hypotheses. 1) To examine if IL-33 protects the liver in acute hepatitis and persistent viral infection. In this Aim, we will examine he endogenous levels of IL-33 and its receptor ST2 in the liver and serum following i.v. delivery of recombinant adenovirus (rAd). To investigate the IL-33/ST2 pathway in persistent viral infection, we will also examine hepatitis induced by lymphocytic choriomeningitis virus clone 13 (LCVM CL13) and antiviral immune responses in these animals. The function of IL-33 will be assessed in IL-33-/- mice as well as by anti-ST2 mAb in wild-type (wt) animals. Conversely, we will treat infected animals with rIL-33 to evaluate its protective functions. 2) To analyze IL-33-induced immune mechanisms responsible for enhanced immune responses and ameliorated liver injury. Our working hypothesis is that IL-33 can directly act on T effector cells or through inducing ILC2s in the liver. We will test the possibility that IL-33 not only enhances T cell vigor but also expands its antigenic breadth using a reconstituted LCMV TCR transgenic mouse (P14) model. In preliminary studies, we found that IL-33 can enhance IFN-? but inhibit TNF-? production. We hypothesize that pro-IL-33, an unprocessed nucleoprotein, can act as a transcriptional factor to modulate the effector cytokines expression on T cells. To examine the IL-33 and DNA interaction, we will use a two-step ChIP assay to determine whether IL-33 binds and sequesters NF-?B signal for TNF-? expression. We anticipate that lack of IL-33 in the knockout animals will lead NF-?B signaling and TNF-? dysregulation, leading to exacerbated liver injury. The new evidence from this study may explain clinical observations that damage-associated molecules like IL-33 and its receptor correlates to T responses and clinical prognosis in patients with chronic hepatitis and drug-induced hepatotoxicity. Such findings would be potentially paradigm-shifting. Although this research is exploratory in nature, it has strong potential to be developed to more detailed studies involving future therapeutic candidates and human subjects infected with hepatitis virus.

描述（由申请人提供）：调节病毒性肝炎中免疫控制与疾病发病机制的关键分子事件仍然难以捉摸。然而，最近的报告表明，慢性肝炎患者血清中警报素（IL-33）及其诱饵受体 sST2 浓度升高。它们的水平与肝损伤、病毒和抗病毒治疗的临床结果相关。我们假设 IL-33 可以直接参与肝脏中的先天性和适应性免疫系统，并且还诱导新的第 2 组先天性淋巴细胞 (ILC2) 的分化。通过这些作用，IL-33 可以调节免疫反应并在炎症过程中保护肝脏。提出了两种互补的方法来检验这些假设。 1) 检查IL-33在急性肝炎和持续性病毒感染中是否具有保护肝脏的作用。在这个目标中，我们将检查静脉注射后肝脏和血清中 IL-33 及其受体 ST2 的内源水平。重组腺病毒（rAd）的递送。为了研究持续性病毒感染中的 IL-33/ST2 途径，我们还将检查淋巴细胞性脉络丛脑膜炎病毒克隆 13 (LCVM CL13) 诱导的肝炎和这些动物的抗病毒免疫反应。 IL-33 的功能将在 IL-33-/- 小鼠中以及野生型 (wt) 动物中通过抗 ST2 mAb 进行评估。相反，我们将用rIL-33治疗受感染的动物以评估其保护功能。 2) 分析 IL-33 诱导的免疫机制，增强免疫反应并改善肝损伤。我们的工作假设是 IL-33 可以直接作用于 T 效应细胞或通过诱导肝脏中的 ILC2。我们将测试 IL-33 不仅增强 T 细胞活力，而且还增强 T 细胞活力的可能性。 使用重建的 LCMV TCR 转基因小鼠 (P14) 模型扩大了其抗原宽度。在初步研究中，我们发现IL-33可以增强IFN-α。但抑制TNF-α生产。我们假设 pro-IL-33（一种未加工的核蛋白）可以作为转录因子来调节 T 细胞上效应细胞因子的表达。为了检查 IL-33 和 DNA 相互作用，我们将使用两步 ChIP 测定来确定 IL-33 是否结合并隔离 TNF-α 的 NF-κB 信号。表达。我们预计敲除动物中IL-33的缺乏将导致NF-κB信号传导和TNF-α信号传导。失调，导致肝损伤加剧。这项研究的新证据可以解释临床观察结果，即 IL-33 及其受体等损伤相关分子与慢性肝炎和药物引起的肝毒性患者的 T 反应和临床预后相关。这些发现可能会改变范式。尽管这项研究本质上是探索性的，但它有很大的潜力发展为涉及未来治疗候选者和感染肝炎病毒的人类受试者的更详细的研究。