T-CELL CYTOKINE/ADHESION MOLECULE REGULATION IN ASTHMA

鉴定 T 细胞控制主要 IgE 调节开关通过它们直接与 B 细胞相互作用并通过 可溶性细胞因子白细胞介素 4 (IL-4) 和 最近鉴定出 CD4 T 辅助细胞的体内作用 哮喘的发生使T细胞的反应功能减弱 肛周过敏原或哮喘气道中暴露的过敏原是关键 问题。 该提案的具体目标旨在描述 参与独特细胞因子基因的转录活性因子 在 T 细胞水平上激活，特别是 IL-4，并延长这些 具体目的是检查这些差异表达 T 细胞克隆系中的 T 细胞调节因子 哮喘家庭。 将尝试将描述关联起来 这些新因素在基因水平上与哮喘易感性有关。 具体目标 1 识别和表征转录因子 T 细胞中 IL-4 基因激活所必需的。 在这些实验中我们 将评估和表征从细胞核中提取的蛋白质因子 过敏原特异性 T 细胞系以及过敏原特异性 T 细胞克隆 影响 IL-4 基因转录的能力 我们将检查 这些蛋白质提取物结合启动子片段基序 发现在 IL-4 激活的调节中具有重要作用。 我们会 纯化并最终测序并克隆负责这些的基因 转录因子并开发试剂来研究其表达 在各种情况下。 在具体目标 2 中，将检查 先前鉴定的 IL-4 转录因子的表达 特应性 T 细胞系和克隆的使用。 实际检验 这些过敏原触发细胞对 IL-4 和 IL-5 使用的选择将 也进行，因为这些因素可能会确定特应性的危险因素 和哮喘。 在具体目标 3 中，我们将研究以下表达式： 来自受影响和过敏原的 T 细胞中的转录因子 患有 IgE 升高和哮喘的亲属家庭中未受影响的个体。 在此特定目标中，建议进行实验来详细检查 T 来自个体的转录因子的细胞系表达 对患有特应性和哮喘的未受影响和受影响的家庭成员进行跟踪 长期在国家犹太教。 将对激活进行分析 这些过敏原特异性和反应的概况和差异 控制 T 细胞并尝试将这些发现与 在一个大的哮喘家族中进行哮喘的临床测量将 完成了。 最后，在具体目标 4 中，我们将研究过敏原驱动的 气道和外周血 T 细胞粘附分子的表达 哮喘。 这些研究对于评估潜力非常重要 哮喘患者气道中 T 细胞积聚的机制 特定过敏原的影响。 这些研究将确定潜力 T细胞细胞因子回路在慢性疾病持续过程中的机制 哮喘中的气道炎症。

The identification of T cells as being in control of the major IgE regulatory switch through their interaction directly with B cells and via the elaboration of the soluble cytokine, interleukin-4 (IL-4), and the recent identification of the in vivo role of CD4+T helper cells into generation of asthma make the function of T cells in the response to perianal allergen or exposed allergen in asthmatic airways a critical issue. Specific aims of this proposal look to characterize transcriptionally active factors involved in unique cytokine gene activation, specifically IL-4, at the T cell level and to extend these specific aims to an examination of the differential expression of these T cell regulatory factors at the T cell clone line in kindreds of asthmatic families. An attempt will be made to correlate description of these new factors with susceptibility to asthma at the genetic level. Specific Aim 1 identifies and characterizes transcription factors necessary for IL-4 gene activation in T cells. In these experiments we will assess and characterize protein factors extracted nuclei of allergen-specific T cell lines as well as allergen-specific T cell clones to influence the transcription of IL-4 genes We will examine the ability of these protein extracts to bind promoter fragment motifs that have been found to be important in the regulation of IL-4 activation. We will purify and eventually sequence and clone the genes responsible for these transcription factors and develop reagents to study their expression under various circumstances. In Specific Aim 2 will examine the expression of the previously identified transcription factors for IL-4 usage by atopic T cell lines and clones. An examination into the actual selection of IL-4 and IL-5 usage by these allergen triggered cells will also be conducted since these factors may identify risk factors for atopy and asthma. In Specific Aim 3, we will look at the expression of transcription factors in allergen derived T cells from affected and unaffected individuals in kindred families with elevated IgE and asthma. In this specific aim, experiments are proposed to examine in detail T cell line expression of transcription factors from individuals from unaffected and affected family members with atopy and asthma followed longterm at National Jewish. Analysis will be made of the activation profiles and differences in the response of these allergen specific and control T cell and an attempt to correlate these findings with the clinical measures of asthma in a large kindred of asthmatic families will be done. Finally, in Specific Aim 4 we will examine the allergen-driven expression of airway and peripheral blood T cell adhesion molecules in asthma. These studies will be important to evaluate the potential mechanisms for T cell accumulation in the airways of asthmatics under the influence of specific allergen. These studies will identity potential mechanisms for T cell cytokine circuits in the perpetuation of chronic airways inflammation in asthma.