Unique immune regulation by alternatively spliced interleukin-4

通过选择性剪接的 IL-4 实现独特的免疫调节

• 批准号: 8196305
• 负责人: Sergei P. Atamas
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2014-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196305
• 关键词: 1-Phosphatidylinositol 3-Kinase; Address; Affect; Aging; Allergic; Allergic inflammation; Alveolar wall; Amino Acids; Animal Model; Animals; Architecture; Asthma; Attenuated; Autoimmune Diseases; Autoimmunity; Binding; Biology; Blood; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; CCL2 gene; Cell Count; Cell Culture Techniques; Cell Surface Receptors; Cells; Characteristics; Chronic; Chronic Disease; Chronic Obstructive Airway Disease; Collagen; Complex; DNA Binding; Data; Deposition; Dermatomyositis; Development; Diagnostic; Disease; Dose; Exons; Extrinsic asthma; Functional disorder; Future; Gene Delivery; Gene Expression; General Population; Genes; Goals; Health; Healthcare; Homeostasis; Human; Hypersensitivity; IRS1 gene; IRS2 gene; Immune; Immunity; In Vitro; Inflammation; Inflammatory; Interferon Type II; Interferons; Interleukin 2 Receptor Gamma; Interleukin 4 Receptor; Interleukin-1 alpha; Interleukin-11; Interleukin-13; Interleukin-2; Interleukin-3; Interleukin-4; Interleukins; Interstitial Lung Diseases; Investigation; Leucocytic infiltrate; Liquid substance; Liver; Lung; Lymphocyte; Mediating; Messenger RNA; Military Personnel; Modeling; Molecular; Molecular Profiling; Mus; Organ; Ovalbumin; Pathway interactions; Patients; Pattern; Phenotype; Phosphorylation; Physiology; Play; Population; Predisposition; Process; Production; Proteins; Pulmonary Pathology; RNA Splicing; Regulation; Relative (related person); Research; Rheumatoid Arthritis; Role; STAT6 gene; Scleroderma; Severities; Severity of illness; Signal Pathway; Signal Transduction; Signaling Molecule; Skin; Steroid therapy; Structure of parenchyma of lung; System; T-Lymphocyte; TNF gene; Testing; Therapeutic; Time; Tuberculosis; Tumor Necrosis Factor-alpha; Variant; Veterans; Widespread Disease; airway remodeling; autocrine; base; cytokine; human TNF protein; improved; in vivo; mRNA Expression; mouse model; neuronal cell body; novel; novel strategies; overexpression; peripheral blood; protein expression; public health relevance; receptor; research study; socioeconomics; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): Project Summary Interleukin (IL)-4 plays a central role in the regulation of immune homeostasis and in various immune-mediated diseases including but not limited to asthma, allergies, tuberculosis, and autoimmunity-associated interstitial lung disease (scleroderma, rheumatoid arthritis, poly- and dermatomyositis). We recently described a splice variant of IL-4, so-called IL-442. The biology of IL-442 appears to be very different from that of IL-4, however the functions of this variant are still poorly understood. We found that the expression levels of IL-442 are dramatically increased in various diseases, including in asthma, to levels that are similar to or exceed those of IL-4. Furthermore, we recently found that IL-442 changes expression levels of numerous genes in human T cells, and that the IL-442-induced changes in the expression profile differ from those induced by IL-4. Unlike IL- 4, IL-442 does not stimulate phosphorylation of STAT6 in a broad range of tested concentrations. Similar to IL- 4, IL-442 stimulates phosphorylation of Jak1, Jak3, and Tyk2 in T cells in a time- and dose-dependent fashion. Gene delivery of IL-442 to mouse lungs causes proinflammatory changes in pulmonary milieu, with the induced levels of TNF-1, IL-11, IFN-3, IL-12p40, and MCP-1 significantly exceeding those induced by gene delivery of IL-4. In bronchoalveolar lavage and blood T cells of patients with asthma, relative expression levels of IL-442 were in many cases higher than those of IL-4. We also discovered that IL-442 potently stimulates production of MCP-1 in human T cells and even more potently (by several hundred fold) upregulates IL-442 production in autocrine fashion. Based on our preliminary data, the Specific Hypothesis of this study is that IL-442 binds to a specific cell surface receptor on T cells; activates characteristic intracellular signaling that is different from signaling induced by IL-4; changes gene expression in a fashion different from that induced by IL-4, particularly stimulating its own production in autocrine fashion; and ultimately stimulates production of numerous proinflammatory and Th1 molecules. Furthermore, the hypothesis is that IL-442 causes significant proinflammatory changes in the lungs in vivo, and is associated with more severe asthma in human patients. In Specific Aim 1, the effects of IL-442 on primary human lymphocytes, as well the regulation of its production at the molecular level, will be investigated in cell culture. In Specific Aim 2, effects of IL-442 gene delivery in vivo will be investigated in normal mice as well as in an ovalbumin-sensitized mouse model of asthma. In Specific Aim 3, the association of IL-442 with disease severity in asthma patients will be studied. This study is directly relevant to Veterans Healthcare. Many of the IL-4-mediated diseases have no known cures and are widely spread in the general population and particularly in veterans. These diseases are chronic, severe, debilitating, and even deadly. The strengths of this investigation are that it addresses a novel topic in a mechanistic way; that it combines mechanistic studies in cell culture, in experimental animals, and in human patients; and that it deals with a molecule that is involved in the mechanisms of numerous diseases that are prevalent in veterans. This study is expected to generate a wealth of novel information on IL-442 as a potential diagnostic marker as well as important target for future therapeutic modulation of numerous diseases in veterans. PUBLIC HEALTH RELEVANCE: Production of a body molecule called interleukin-4 (IL-4) can be elevated and drive disease processes in the lungs, liver, skin, and other organs. All of these IL-4-dependent chronic diseases may develop with aging or as a consequence of military exposures, thus making them prevalent in the veteran population. We recently discovered that IL-4 is made in two forms, traditional IL-4 and a form called IL-4-delta-2 (IL-442). We discovered that this other form is excessively produced in patients and acts differently from the traditional IL-4 on the cells of the body. The proposed research will look at the effects of IL-442 on the cells isolated from humans, investigate the disease caused by excess IL-442 in mouse lungs, and determine how excess IL-442 in humans is associated with disease severity. The results will form the basis for development of new therapies that target IL-442 in patients, particularly in veterans, with the goal of attenuating or curing diseases in veterans, and thus improving veterans' health.

描述（由申请人提供）： 项目摘要 白细胞介素 (IL)-4 在免疫稳态调节和各种免疫介导的疾病中发挥着核心作用，包括但不限于哮喘、过敏、结核病和自身免疫相关的间质性肺病（硬皮病、类风湿性关节炎、多发性关节炎） - 和皮肌炎）。我们最近描述了 IL-4 的剪接变体，即所谓的 IL-442。 IL-442 的生物学似乎与 IL-4 非常不同，但对该变体的功能仍知之甚少。我们发现，IL-442 的表达水平在各种疾病（包括哮喘）中显着增加，达到与 IL-4 相似或超过的水平。此外，我们最近发现IL-442改变了人类T细胞中许多基因的表达水平，并且IL-442诱导的表达谱变化与IL-4诱导的表达谱变化不同。与IL-4不同，IL-442在广泛的测试浓度范围内不会刺激STAT6的磷酸化。与 IL-4 类似，IL-442 以时间和剂量依赖性方式刺激 T 细胞中 Jak1、Jak3 和 Tyk2 的磷酸化。将 IL-442 基因递送至小鼠肺部会引起肺环境促炎变化，诱导的 TNF-1、IL-11、IFN-3、IL-12p40 和 MCP-1 水平显着超过 IL 基因递送诱导的水平-4。在哮喘患者的支气管肺泡灌洗液和血液T细胞中，IL-442的相对表达水平在许多情况下高于IL-4的相对表达水平。我们还发现 IL-442 有效刺激人类 T 细胞中 MCP-1 的产生，甚至更有效（数百倍）以自分泌方式上调 IL-442 的产生。根据我们的初步数据，本研究的具体假设是 IL-442 与 T 细胞上的特定细胞表面受体结合；激活与 IL-4 诱导的信号传导不同的特征性细胞内信号传导；以不同于 IL-4 诱导的方式改变基因表达，特别是以自分泌方式刺激其自身产生；并最终刺激大量促炎分子和 Th1 分子的产生。此外，假设 IL-442 在体内引起肺部显着的促炎变化，并且与人类患者更严重的哮喘有关。在具体目标 1 中，将在细胞培养中研究 IL-442 对原代人淋巴细胞的影响，以及在分子水平上对其产生的调节。在具体目标 2 中，将在正常小鼠以及卵清蛋白致敏的哮喘小鼠模型中研究 IL-442 基因递送的体内效果。在具体目标 3 中，将研究 IL-442 与哮喘患者疾病严重程度的关系。这项研究与退伍军人医疗保健直接相关。许多 IL-4 介导的疾病尚无已知的治疗方法，并且在普通人群中广泛传播，特别是在退伍军人中。这些疾病是慢性的、严重的、使人衰弱的，甚至是致命的。这项研究的优点在于它以机械的方式解决了一个新颖的话题；它结合了细胞培养、实验动物和人类患者的机制研究；它涉及一种与退伍军人中常见的多种疾病机制有关的分子。这项研究预计将产生大量关于 IL-442 的新信息，作为潜在的诊断标记物以及退伍军人多种疾病的未来治疗调节的重要靶点。 公共卫生相关性： 体内一种称为白细胞介素 4 (IL-4) 的分子的产生量会增加，并导致肺、肝脏、皮肤和其他器官的疾病进程。所有这些 IL-4 依赖性慢性疾病都可能随着衰老或军事暴露而发生，从而使它们在退伍军人群体中普遍存在。我们最近发现 IL-4 有两种形式，传统的 IL-4 和称为 IL-4-delta-2 (IL-442) 的形式。我们发现这种另一种形式在患者体内过量产生，并且对身体细胞的作用与传统的 IL-4 不同。拟议的研究将着眼于 IL-442 对从人体中分离出的细胞的影响，研究小鼠肺部中过量的 IL-442 引起的疾病，并确定人体中过量的 IL-442 与疾病严重程度之间的关系。这些结果将为开发针对患者（特别是退伍军人）的 IL-442 新疗法奠定基础，目标是减轻或治愈退伍军人的疾病，从而改善退伍军人的健康。