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: 7924922
• 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/7924922
• 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; 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）： 项目摘要介绍（IL）-4在免疫体内稳态和各种免疫介导的疾病中起着核心作用，包括但不限于哮喘，过敏，结核病和自身免疫性相关的间质肺肺疾病（Sclerodersial，Sclerodersima，Scleroderma，Rheamumapoid Arthrthiation，Poly-Poly-somyossisision）。我们最近描述了IL-4，所谓的IL-442的剪接变体。 IL-442的生物学似乎与IL-4的生物学有很大不同，但是该变体的功能仍然很少了解。我们发现，包括哮喘在内的各种疾病中，IL-442的表达水平急剧增加到与IL-4相似或超过IL-4的水平。此外，我们最近发现IL-442改变了人T细胞中许多基因的表达水平，并且IL-442诱导的表达谱的变化与IL-4诱导的表达谱的变化不同。与IL-4不同，IL-442不会在广泛的测试浓度中刺激STAT6的磷酸化。与IL-4相似，IL-442以时间和剂量依赖性的方式刺激T细胞中JAK1，JAK3和TYK2的磷酸化。 IL-442至小鼠肺的基因递送会导致肺部环境的促炎变化，诱导TNF-1，IL-11，IFN-3，IL-3，IL-12P40和MCP-1的水平显着超过了IL-4基因递送引起的IL-4诱导的水平。在哮喘患者的支气管肺泡灌洗和血液T细胞中，在许多情况下，IL-442的相对表达水平高于IL-4。我们还发现，IL-442在人类T细胞中有效刺激MCP-1的产生，甚至更有效地（数百倍）以自分泌方式上调IL-442的产生。基于我们的初步数据，这项研究的特定假设是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细胞内信号传导；以与IL-4引起的方式不同的方式改变基因表达，特别是以自分泌方式刺激其自身的生产；并最终刺激了许多促炎和Th1分子的产生。此外，该假设是IL-442在体内引起肺部的显着促炎性变化，并且与人类患者更严重的哮喘有关。在特定的目标1中，IL-442对原代人淋巴细胞的影响以及其在分子水平的产生的调节将在细胞培养中进行研究。在特定的目标2中，将在正常小鼠以及卵蛋白敏感的哮喘模型中研究体内IL-442基因递送的作用。在特定的目标3中，将研究IL-442与哮喘患者疾病严重程度的关联。这项研究与退伍军人医疗保健直接相关。许多IL-4介导的疾病没有已知的治疗方法，并且在普通人群中，尤其是在退伍军人中广泛传播。这些疾病是慢性，严重，使人衰弱甚至致命的。这项调查的优势在于它以机械方式解决了一个新的话题。它结合了细胞培养，实验动物和人类患者的机械研究；并且它涉及与退伍军人普遍存在的许多疾病机制有关的分子。预计这项研究将产生有关IL-442的大量新信息，作为潜在的诊断标记，以及对退伍军人多种疾病的未来治疗调节的重要目标。 公共卫生相关性： 可以升高称为白介素4（IL-4）的人体分子的产生，并在肺，肝，皮肤和其他器官中驱动疾病过程。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展，从而使它们在退伍军人人口中普遍存在。我们最近发现，IL-4以两种形式制成，传统的IL-4和一种称为IL-4-DELTA-2（IL-442）的形式。我们发现这种另一种形式在患者中产生过多，并且与人体细胞上的传统IL-4的作用不同。拟议的研究将研究IL-442对从人类分离的细胞的影响，研究由小鼠肺中过量IL-442引起的疾病，并确定人类中过量的IL-442与疾病严重程度有关。结果将构成针对患者，尤其是退伍军人的IL-442的新疗法开发的基础，其目的是减弱或治愈退伍军人的疾病，从而改善退伍军人的健康。