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，最近的报道表明，慢性肝炎患者血清中ALAMIN(IL-33)及其诱骗受体Sst2浓度升高。它们的水平与肝损伤、病毒感染和抗病毒治疗的临床结果有关。我们假设IL-33可以直接参与肝脏的天然免疫系统和获得性免疫系统，也可以诱导新的第2组天然淋巴样细胞(ILC2)的分化。通过这些作用，IL-33可以调节免疫反应，在炎症过程中保护肝脏。本文提出了两种互补的方法来检验这些假设。1)检测IL-33在急性肝炎和持续性病毒感染中是否对肝脏有保护作用。为此，我们将检测静脉注射后肝脏和血清中IL-33及其受体ST2的内源性水平。重组腺病毒(Rad)的传递。为了研究IL-33/ST2途径在持续病毒感染中的作用，我们还将检测淋巴细胞性脉络膜脑膜炎病毒克隆13(LCVMCL13)诱导的肝炎以及这些动物的抗病毒免疫反应。IL-33的功能将在IL-33-/-小鼠中进行评估，并在野生型(Wt)动物中通过抗ST2单抗进行评估。相反，我们将用rIL-33治疗感染的动物，以评估其保护功能。2)分析IL-33诱导的免疫应答增强和肝损伤减轻的免疫机制。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来发挥作用。我们将测试IL-33不仅增强T细胞活力的可能性，而且 利用重组的LCMV TCR转基因小鼠(P14)模型扩大其抗原广度。在初步研究中，我们发现IL-33可以增强干扰素？但抑制肿瘤坏死因子-？制作。我们假设，未经处理的核蛋白Pro-IL-33可以作为转录因子来调节T细胞上效应细胞因子的表达。为了检测IL-33与DNA的相互作用，我们将使用两步芯片法来确定IL-33是否结合和隔离了肿瘤坏死因子-β的核因子-B信号。表情。我们预计，在基因敲除的动物中，IL-33的缺失将导致核因子-β信号转导和肿瘤坏死因子-？调节失调，导致肝脏损伤加剧。这项研究的新证据可能解释临床观察到的损伤相关分子，如IL-33及其受体与慢性肝炎和药物肝毒性患者的T反应和临床预后相关。这样的发现可能会改变范式。虽然这项研究是探索性的，但它有很大的潜力发展成更详细的研究，涉及未来的治疗候选人和感染肝炎病毒的人类受试者。