ISOCYANATE ANTIGENS AND T CELLS THAT CAUSE ASTHMA

异氰酸酯是一组高活性的广泛使用的低分子 重化学品，是最常见的职业报告的原因 发达国家的哮喘。然而，异氰酸酯 因为哮喘并没有很好的定义。根据我们的初步数据， 在文献中报道，我们假设异氰酸酯诱导的哮喘 依赖于异氰酸酯抗原驱动的T细胞介导的气道 炎症为了解决这一假设，我们建议调查 异氰酸酯诱导哮喘依赖于异氰酸酯抗原驱动的T细胞 介导的气道炎症。为了解决这个问题，我们建议 研究异氰酸酯抗原驱动体外T细胞应答， 在体内暴露后。我们的调查将确定和 表征异氰酸酯抗原和反应性人T细胞。我们将 比较主要暴露部位的异氰酸酯抗原反应性T细胞 (skin/肺），与来自血液的那些，以评估潜在的途径， 异氰酸酯致敏和鉴别诊断指标 敏感性/易感性研究将在人群中进行 暴露于HDI的汽车修理工人的死亡率。患者 将通过与正在进行的实地合作获取样本， 流行病学和临床研究。这项建议的结果将 提供证据来支持或反驳我们的假设，异氰酸酯 哮喘由异氰酸酯抗原特异性T细胞介导， 作为其他类型职业性哮喘的模型。我们特别 目的：1）制备和表征HDI抗原，包括; 异氰酸酯代谢物和异氰酸酯结合物 外源蛋白目的2）评价HDI的T细胞抗原性 抗原，基于血液和肺淋巴细胞增殖。评价 基于血液和肺的HDI抗原的T细胞抗原性 淋巴细胞增殖、细胞因子产生和表型，以便 确定启动气道细胞因子的HDI的分子形式 生产和表型，以确定HDI的分子形式 引发哮喘患者的气道炎症。目标3）建立T- 来自HDI哮喘的皮肤、肺和外周血的细胞系 患者并表征表型、抗原特异性、细胞因子 这些细胞中异氰酸酯应答性T细胞的产生和TCR表达 不同的隔间目的4）比较发现的异氰酸酯响应性T细胞 在皮肤，肺和血液中，并与临床敏感性相关， 确定与暴露和致敏相关的特征 导致临床哮喘。这些研究应有助于确定 生物学相关的HDI抗原，并大大提高我们的 了解异氰酸酯如何启动特定的T细胞反应， 导致异氰酸酯敏感性和哮喘。异氰酸酯的表征 抗原和异氰酸酯反应性T细胞将识别靶点， 异氰酸酯哮喘的诊断、预防和治疗。

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.