REGULATION OF IL4 IN CELLS INVOLVED IN ALLERGIC DISEASE

过敏性疾病细胞中 IL4 的调节

• 批准号: 2371907
• 负责人: VINCENZO CASOLARO
• 金额: $ 11.52万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2371907
• 关键词: basophils; gel mobility shift assay; gene expression; helper T lymphocyte; human tissue; hypersensitivity; immunofluorescence technique; immunoglobulin E; interleukin 4; nuclear factor kappa beta; protein biosynthesis; tissue /cell culture; transcription factor

Allergic diseases affect 30% of the U.S. population. The cytokine interleukin (lL)-4 plays a central role in allergic responses by inducing lgE synthesis and by stimulating several aspects of the inflammatory response. Synthesis of lL-4 is increased in allergic subjects and overexpression of lL-4 in mice leads to the development of "allergic like" inflammatory disease. Production of IL-4 was originally described from T helper (Th) lymphocytes. Indeed, analysis of cytokine profiles was used to define three functional subpopulations of Th cells: ThI, producing lL-2 and interferon v; Th2, producing IL-4 and lL-5; and Th0, having a mixed phenotype. More recently, however, it has been shown that cells other than Th2/Th0 can produce IL-4. For example, human peripheral blood basophils release substantial amounts of lL-4 and other cytokines in response to Crosslinking of membrane-bound lgE. Several studies suggest an important role of lL-4 producing Th cells and basophils in the pathogenesis of allergic inflammation. Delineating the molecular basis of lL-4 gene activation is, therefore, of central importance to our understanding of allergic diseases. The mechanism(s) responsible for mutually exclusive cytokine expression in Th cell subsets are not fully understood. Moreover, no studies have been conducted so far to analyze the transcriptional activation of the lL4 gene in human basophils. The principal investigator has studied in detail two factors having opposite effects on transcriptional activation of the lL-2 and IL-4 genes: nuclear factor (NF)KB and CP2. This project is focused on two specific aims: 1) to understand the mechanisms of ILA gene regulation by these factors and their role in differential gene expression in human Th1 and Th2 clones, and 2) to investigate the mechanisms of lL-4 gene regulation in human basophils, to identify possible transcriptional activators mediating lgE-dependent ILA expression in these cells. DNA-binding assays with proteins extracted from human ragweed antigen (Amb a I)-specific Th1 and Th2 clones and freshly isolated, purified basophils and the analysis of IL4 promoter-driven reporter gene expression in the human T cell leukemia Jurkat will be used to accomplish these aims. Defining the transcriptional factors that regulate the expression of the human ILA gene in Th cells and basophils may, in the long term, lead to novel therapeutic approaches for the treatment of allergic disorders.

过敏性疾病影响了30%的美国人口。细胞因子 白细胞介素（IL）-4在变态反应中起着中心作用， 诱导IgE合成，并通过刺激几个方面， 炎症反应。IL-4的合成在过敏反应中增加， 受试者和IL-4在小鼠中的过表达导致发展 “过敏样”炎症性疾病的症状IL-4的产生是 最初描述自T辅助（Th）淋巴细胞。的确， 细胞因子谱的分析用于定义三种功能性细胞因子。 Th细胞亚群：ThI，产生IL-2和干扰素V; Th 2，产生IL-4和IL-5;和Th 0，具有混合表型。 然而，最近的研究表明， Th 2/Th 0可产生IL-4。例如，人类外周血 嗜碱性粒细胞释放大量IL-4和其他细胞因子， 对膜结合IgE的交联的响应。几项研究 提示产生IL-4 Th细胞和嗜碱性粒细胞的重要作用 过敏性炎症的发病机制。划定 因此，IL-4基因激活的分子基础是 对我们了解过敏性疾病的重要性。 负责互斥细胞因子的机制 Th细胞亚群中的表达尚未完全了解。更没有 到目前为止，已经进行了一些研究来分析转录 人嗜碱性粒细胞中IL 4基因的激活。校长 研究人员详细研究了两个具有相反影响的因素 IL-2和IL-4基因的转录激活：核 因子（NF）KB和CP 2。该项目重点关注两个具体的 目的：1）了解ILA基因的调控机制， 这些因子及其在人类基因表达差异中的作用 Th 1和Th 2克隆，以及2）研究IL-4的作用机制， 人类嗜碱性粒细胞的基因调控，以确定可能的 转录激活因子介导IgE依赖性ILA表达， 这些细胞。人源蛋白质的DNA结合试验 豚草抗原（Amb a I）特异性Th 1和Th 2克隆和新鲜的 分离、纯化的嗜碱性粒细胞和IL 4启动子驱动的 报告基因在人T细胞白血病Jurkat中的表达将是 用于实现这些目标。转录因子的定义 调节Th细胞中人ILA基因的表达， 从长远来看，嗜碱性粒细胞可能会导致新的治疗方法， 用于治疗过敏性疾病。