Neuroimmune modulation across the BBB: Regulation of ILS trafficking by TNF

跨 BBB 的神经免疫调节：TNF 对 ILS 运输的调节

• 批准号: 8133699
• 负责人: Weihong Pan
• 金额: $ 31.51万
• 依托单位: LSU PENNINGTON BIOMEDICAL RESEARCH CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8133699
• 关键词: AIDS neuropathy; Acute Transverse Myelitis; Address; Age of Onset; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Autoimmune Diseases; Bacterial Infections; Behavior; Biological Assay; Blood; Blood - brain barrier anatomy; Brain; Brain Pathology; CNS autoimmune disease; Cell Culture Techniques; Central Nervous System Diseases; Cerebrum; Clinical; Cytokine Signaling; Data; Diabetes Mellitus; Diagnosis; Disease; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Etiology; Exhibits; Experimental Autoimmune Encephalomyelitis; Feasibility Studies; Feline Panleukopenia; Gliosis; Goals; Health; Hypoxia; IL2RA gene; Immunohistochemistry; In Situ; Inflammatory; Interleukin-15; Intervention; Knock-out; Knockout Mice; Ligands; Lipopolysaccharides; Literature; Mediating; Mediator of activation protein; Metabolic syndrome; Minor; Modeling; Multiple Sclerosis; Mus; Neurodegenerative Disorders; Organelles; Outcome; Parkinson Disease; Pathway interactions; Perfusion; Peripheral; Permeability; Physicians; Play; Production; Prostaglandins; Protein Biosynthesis; Proteins; Regulation; Resolution; Role; Signal Transduction; Sorting - Cell Movement; Stroke; Surface; Symptoms; System; TNF gene; Testing; Time; Transcriptional Activation; Translations; Treatment Efficacy; Tumor Necrosis Factor-alpha; United States; Up-Regulation; Viral; Work; burden of illness; cyclooxygenase 2; cytokine; encephalomyelopathy; experience; neuroinflammation; new therapeutic target; novel; protein degradation; receptor; trafficking; transcytosis; tumor

DESCRIPTION (provided by applicant): The goals of the proposed study are to determine the mechanisms by which TNF, an important component of neuroinflammation, upregulates IL15 production in the endothelial cells of the blood-brain barrier (BBB), the transport of IL15 across the BBB, and the resulting pathophysiological implications for neuroinflammatory and autoimmune disorders. The proinflammatory cytokine TNF not only crosses the BBB to exert its multipotent effects, but it also modulates the endothelial signaling and permeation of other cytokines. IL15 and its receptors show specific and robust upregulation in cerebral microvessel endothelial cells challenged by TNF or lipopolysaccharide, shown by microarray, qPCR and immunohistochemistry in our preliminary results. We will address the overall hypothesis that IL15 potentiates the effects of TNF in the CNS with four aims. (1) Test the hypothesis that TNF upregulates IL15 transport. We will identify the roles of IL15 receptors in transporting IL15 by use of in-situ brain perfusion studies in receptor knockout and wildtype mice, determine the effects of TNF on the transcellular permeability of IL15 in primary cerebral microvessel endothelial cells cultured in the Transwell system, and determine the differential roles and interactions of IL15R1, IL2R2, and IL2R3 receptors in TNF-facilitated IL15 transcytosis by immunofluorescent trafficking studies. (2) Test the hypothesis that TNF stimulates protein synthesis and basolateral secretion of IL15 from the endothelial cells of the BBB. This will be achieved by protein translation and turnover assays, confocal analyses of time-dependent colocalization of IL15, its receptors, and intracellular organelles, and quantification by enzyme-linked immunosorbent assay. We will also test the mediating effect of prostaglandins in the TNF-induced increase of the basolateral secretion of IL15. (3) Test the hypothesis that an upregulated endothelial IL15 system enhances TNF-induced neuroinflammation, shown by fever, sickness behavior, reactive gliosis, and cerebral production of proinflammatory cytokines. Apart from studies on mice to test the effects of soluble IL15 receptors in blocking these effects, the inflammatory changes in cultured cerebral microvessel endothelial cells obtained from knockout and wildtype mice will also be identified. (4) Test the hypothesis that an upregulated IL15 system exacerbates experimental autoimmune encephalomyelitis (EAE), a disorder of neuroimmune modulation and a model for multiple sclerosis. The regulatory changes of IL15 permeation across the BBB of the EAE mice will be determined, and the effects of soluble receptors in reducing the symptoms and disease burden of EAE will be determined. The feasibility of the studies is supported by preliminary results, our past experience, and the literature. By completion of the proposed studies, we will acquire novel and essential understanding of IL15 and IL15 receptor trafficking in cerebral endothelial cells, and identify new therapeutic targets for intervention in neuroinflammation and CNS autoimmune diseases. PUBLIC HEALTH RELEVANCE Neuroinflammation is a common etiology in many CNS disorders, including infection (bacterial, fungal, viral, and parasitic), neuroAIDS, stroke, global hypoxia, autoimmune diseases (e.g., multiple sclerosis, transverse myelitis, acute disseminating encephalomyelopathy), tumor, a variety of neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis), and even diabetes and the metabolic syndrome. Multiple sclerosis (MS) is a devastating and progressive disorder with an age of onset typically between 20 and 40. There are about 250,000 - 350,000 people in the United States with MS diagnosed by a physician. This estimate suggests that about 200 new cases are diagnosed each week. The proinflammatory cytokines TNF and IL15 play important roles in the initiation, progression, and resolution of neuroinflammation and autoimmune disorders of the CNS. Their interactions with the blood-brain barrier largely determine the final outcome of these blood-borne cytokines.

描述（由申请人提供）：拟议的研究的目标是确定TNF（神经炎症的重要组成部分）上调的机制，上调了血脑屏障（BBB）内皮细胞中IL15的产生（BBB），IL15的运输，BBB跨BBB的运输，以及导致的病理生理学症状和自动源自病原体。促炎细胞因子TNF不仅越过BBB以发挥其多能效应，而且还调节了其他细胞因子的内皮信号传导和渗透。 IL15及其受体显示出受TNF或脂多糖挑战的脑微血管内皮细胞的特异性和强大上调，在我们的初步结果中，微阵列，QPCR和免疫组织化学显示了。我们将解决以下总体假设，即IL15以四个目标增强了TNF在中枢神经系统中的影响。 （1）检验TNF上调IL15转运的假设。 We will identify the roles of IL15 receptors in transporting IL15 by use of in-situ brain perfusion studies in receptor knockout and wildtype mice, determine the effects of TNF on the transcellular permeability of IL15 in primary cerebral microvessel endothelial cells cultured in the Transwell system, and determine the differential roles and interactions of IL15R1, IL2R2, and通过免疫荧光贩运研究，TNF-纤维化IL15的IL2R3受体。 （2）检验了TNF刺激BBB内皮细胞中IL15的蛋白质​​合成和基底外侧分泌的假设。这将通过蛋白质翻译和周转测定，IL15的时间依赖性共定位，其受体和细胞内细胞器以及通过酶联免疫吸附测定法进行定量来实现。我们还将测试前列腺素在TNF诱导的IL15基底外侧分泌增加中的介导作用。 （3）检验了上调内皮IL15系统可增强TNF诱导的神经炎症的假设，该神经炎症是由发烧，疾病行为，反应性神经病和促炎细胞因子产生的大脑产生的。除了对小鼠的研究测试可溶性IL15受体在阻断这些作用方面的作用外，还将鉴定出从敲除和野生型小鼠获得的培养的脑微血管内皮细胞的炎症变化。 （4）检验以下假设：上调的IL15系统加剧了实验性自身免疫性脑脊髓炎（EAE），神经免疫调节的疾病以及用于多发性硬化症的模型。将确定跨EAE小鼠BBB的IL15渗透的调节变化，并确定可溶性受体在减轻EAE症状和疾病负担中的影响。这些研究的可行性得到了初步结果，我们过去的经验和文献的支持。通过完成拟议的研究，我们将在脑内皮细胞中对IL15和IL15受体运输的新颖和基本理解，并确定用于干预神经炎症和CNS自身免疫性疾病的新的治疗靶标。公共卫生相关性神经炎症是许多中枢神经系统疾病的常见病因，包括感染（细菌，真菌，病毒和寄生虫），神经辅助，中风，全球缺氧，自身免疫性疾病，自身免疫性疾病（例如，多发性硬膜病，横向骨髓炎，急性骨髓炎，急性骨膜化的杂种症，均具有差异化的杂种症，是素质的杂种，敏感性，差异化，促成了差异，这些疾病是变性的。 （例如，阿尔茨海默氏病，帕金森氏病，肌萎缩性侧索硬化症），甚至糖尿病和代谢综合征。多发性硬化症（MS）是一种毁灭性和进行性疾病，发病年龄通常在20到40岁之间。该估计表明，每周诊断出约200例新病例。促炎细胞因子TNF和IL15在CNS神经炎症和自身免疫性疾病的启动，进展和分辨率中起着重要作用。他们与血脑屏障的相互作用在很大程度上决定了这些血源性细胞因子的最终结果。