Cellular auto-immune mechanisms of narcolepsy

发作性睡病的细胞自身免疫机制

• 批准号: 9404243
• 负责人: Igor J Koralnik
• 金额: $ 27.13万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9404243
• 关键词: Cellular; auto; immune; mechanisms; narcolepsy

Narcolepsy with Cataplexy (N/C) is a common sleep disorder, but very little is understood about the underlying mechanism that causes N/C and why symptoms are severe in some patients but not in others. N/C is caused by selective loss of the hypothalamic neurons that produce the orexin neuropeptides and is strongly associated with the Major Histocompatibility Complex (MHC) class II allele DQB1 *0602. We hypothesize that N/C is caused by CD4+ or CD8+ T cells response directed against the hypothalamic neurons producing the orexin neuropeptides, and that a more aggressive immune response results in increased severity of N/C. This hypothesis is supported by a number of publications suggesting a causal role of the cellular immune response in N/C as well as our own preliminary data. Our long term goal is to understand the cause of N/C and the determinants of disease severity. In doing so, we will also devise a much needed blood test for the early diagnosis of NC and prediction of disease severity. Our objectives are to determine whether N/C patients mount a cellular immune response to orexin peptides and whether this response is associated with more severe symptoms. The rationale is that these proposed studies will enable us to develop a blood test for early diagnosis of N/C and predictor of disease severity. To test these hypotheses, we will pursue the following set of Specific Aims: Aim 1) Correlate disease severity with the cellular immune response to orexin neuropeptides in DQB1 *0602+ Narcolepsy with Cataplexy patients. We will measure narcolepsy symptoms severity using well- validated scales and correlate symptom severity with the reactivity of CD4+ or CD8+ T cells to orexin peptides using three different assays: 1) Intracellular Cytokine Staining (ICS) assay 2) Luminex xMAP bead array on cell culture supernatant and 3) RT-qPCR for cytokine mRNA on cellular RNA . Aim 2) Identify immunodominant orexin epitopes recognized by CD4+ or CD8+ T cells in DQB1 *0602+ Narcolepsy with Cataplexy patients. We will map orexin epitopes in N/C patients using ICS and devise a diagnostic blood test and predictor of disease severity for N/C. The approach is innovative, because it departs significantly from the status quo by focusing on the cellular mechanisms leading to N/C. The proposed research is significant because it will allow us to identify the immunopathogenic mechanisms leading to N/C, develop a blood test for early diagnosis of N/C and predict disease severity. The knowledge gained from these studies will also have far reaching implications in advancing the fields of Sleep Disorders and Neuro-Immunology.

发作性的发作性疾病（N/C）是一种常见的睡眠障碍，但对基础知识几乎没有理解 引起N/C的机制以及某些患者的症状很严重而在其他患者中却不严重。 n/c是引起的 通过选择性丧失的下丘脑神经元产生Orexin neuropypides，并且密切相关 具有主要的组织相容性复合物（MHC）II类等位基因DQB1 *0602。我们假设N/C是 由CD4+或CD8+ T细胞引起的针对下丘脑神经元的反应，产生Orexin 神经肽，并且更具侵略性的免疫反应导致N/C的严重程度增加。这 许多出版物支持假设，表明细胞免疫反应的因果作用 在N/C以及我们自己的初步数据中。我们的长期目标是了解N/C的原因 疾病严重程度的决定因素。这样，我们还将为早期设计急需的血液检查 诊断NC和疾病严重程度的预测。我们的目标是确定N/C患者是否 安装对Orexin肽的细胞免疫反应，以及该反应是否与更多有关 严重的症状。理由是这些拟议的研究将使我们能够早期进行血液检查 N/C的诊断和疾病严重程度的预测指标。 为了检验这些假设，我们将追求以下一组具体目标： 目标1）将疾病的严重程度与DQB1中的细胞免疫反应相关 *0602+与瘫痪的患者的发肠疾病。我们将使用良好的 经过验证的量表并将症状的严重程度与CD4+或CD8+ T细胞的反应性与Orexin肽的反应性相关联 使用三种不同的测定：1）细胞内细胞因子染色（ICS）测定2）luminex xmap珠子阵列在细胞上 培养上清液和3）RT-QPCR用于细胞因子RNA上的细胞因子mRNA。 AIM 2）确定DQB1 *0602+ CD4+或CD8+ T细胞识别的免疫主导OREXIN表位 发作性患者的发肠病毒。我们将使用ICS绘制N/C患者的Orexin Eptopes并设计A 诊断血液检查和N/C疾病严重程度的预测指标。 该方法具有创新性，因为它通过关注细胞来显着脱离现状 导致N/C的机制。拟议的研究很重要，因为它将使我们能够确定 导致N/C的免疫致病机制，开发用于早期诊断的血液测试，并预测 疾病的严重程度。从这些研究中获得的知识也将具有很大的影响 推进睡眠障碍和神经免疫学领域。