ISOCYANATE ANTIGENS AND T CELLS THAT CAUSE ASTHMA

异氰酸酯是一类高活性、用途广泛的低分子化合物 超重化学品，是最常见的职业原因 发达国家的哮喘。然而，异氰酸酯的作用机理 因为哮喘的定义不是很清楚。根据我们的初步数据， 在文献报道中，我们假设异氰酸酯诱发哮喘 依赖于异氰酸酯抗原驱动的T细胞介导的呼吸道 发炎。为了解决这一假设，我们建议调查 异氰酸酯诱导的哮喘依赖于异氰酸酯抗原驱动的T细胞 介导的，呼吸道炎症。为了解决这一假设，我们建议 研究异氰酸酯抗原在体外诱导的T细胞反应 在活体暴露后。我们的调查将确定和 表征异氰酸酯抗原和反应性人类T细胞。我们会 比较初级暴露部位的异氰酸酯抗原反应性T细胞 (皮肤/肺)，与来自血液的那些，以评估潜在的途径 异氰酸酯的敏化和诊断指示剂的确定 敏感性/敏感度。这些研究将在人群中进行 六亚甲基二异氰酸酯(HDI)暴露在车身维修工人手中。有耐心的 样本将通过与正在进行的现场合作来获取 流行病学和临床研究。这项提案的调查结果将 提供证据支持或驳斥我们的假设：异氰酸酯 哮喘是由异氰酸酯抗原特异性T细胞介导的，它还将 作为其他类型职业性哮喘的典范。具体来说，我们 建议：目的1)制备和鉴定HDI抗原，包括； 异氰酸酯代谢物和异氰酸酯偶联的正常人和 外来蛋白质。目的2)评价HDI的T细胞抗原性 抗原，以血液和肺淋巴细胞增殖为基础。评估 基于血液和肺的HDI抗原的T细胞抗原性 淋巴细胞的增殖、细胞因子的产生和表型 鉴定启动呼吸道细胞因子的HDI的分子形式 生产和表型，以确定HDI的分子形式 会引发哮喘患者的呼吸道炎症。目标3)建立T- 高密度脂蛋白哮喘患者皮肤、肺和外周血中的细胞系 并鉴定患者的表型、抗原特异性、细胞因子 异氰酸酯反应性T细胞的产生和TCR的表达 不同的车厢。目的4)比较发现的异氰酸酯反应性T细胞 在皮肤、肺和血液中的含量，并与临床上对 确定与暴露和敏化相关的特征 导致临床哮喘。这些研究应该能够确定 与生物相关的HDI抗原(S)和极大地增强了我们的 了解异氰酸酯如何启动特定的T细胞反应 导致异氰酸酯过敏和哮喘。异氰酸酯的表征 抗原和异氰酸酯反应T细胞将识别靶标 异氰酸酯哮喘的诊断、防治。

Isocyanates are a group of highly reactive widely used low-molecular weight chemicals, and are the most commonly reported cause of occupation asthma in developed countries. Yet, the mechanisms by which isocyanates cause asthma are not well defined. Based on our preliminary data and that reported in the literature, we hypothesize that isocyanate-induced asthma is dependent on isocyanate antigen-driven T-cell mediated, airway inflammation. To address this hypothesis, we propose to investigate isocyanate-induced asthma is dependent on isocyanate antigen-driven T-cell mediated, airway inflammation. To address this hypothesis, we propose to investigate isocyanate antigen-driven T-cell responses in vitro-, following in vivo exposure. Our investigations will identify and characterize isocyanate antigens and reactive human T-cells. We will compare isocyanate antigen-reactive T-cells from primary exposure sites (skin/lung), with those from blood, to evaluate potential routes of sensitization and identify diagnostic indicators of isocyanate sensitivity/susceptibility. The studies will be performed in a population of hexamethylene diisocyanate (HDI) exposed autobody shop workers. Patient samples will be acquired through collaboration with ongoing field epidemiological and clinical studies. The findings of this proposal will provide evidence to support or refute our hypothesis that isocyanate asthma is mediated by isocyanate antigen-specific T-cells and will also serve as a model for other types of occupational asthma. Specifically, we propose to: Aim 1) Generate and characterize HDI antigens including; isocyanate metabolites, and isocyanate conjugated t normal human and foreign proteins. Aim 2) Evaluate the T-cell antigenicity of the HDI antigens, based on blood and lung lymphocyte proliferation. Evaluate the T-cell antigenicity of the HDI antigens, based on blood and lung lymphocyte proliferation, cytokine production, and phenotype in order to identify the molecular form of HDI that initiates airway cytokine production, and phenotype in order to identify the molecular form of HDI that initiates airway inflammation in asthma patients. Aim 3) Establish T- cell lines from the skin, lung and peripheral blood of HDI asthma patients and characterize the phenotype, antigen specificity, cytokine production and TCR expression of isocyanate responsive T-cells in these different compartments. Aim 4) Compare isocyanate responsive T-cells found in the skin, lung and blood and correlate with clinical sensitivity to determine characteristics associated with exposure and sensitization leading to clinical asthma. These studies should enable identification of the biologically relevant HDI antigen(s) and greatly enhance our understanding of how isocyanates initiate specific T-cell responses that lead to isocyanate sensitivity and asthma. Characterization of isocyanate antigens and isocyanate-responsive T-cells will identify targets for the diagnosis, prevention and treatment of isocyanate asthma.