TSLP Signaling in Dendritic Cells - Aim 3

描述(由申请人提供)： 这是德克萨斯医学中心NIH/NIAID哮喘和过敏性疾病合作研究中心的申请，该中心联合德克萨斯大学M.D.安德森癌症中心和贝勒医学院，研究人类胸腺基质淋巴生成素(TSLP)在主导过敏表型的Th2免疫反应的发展和维持中的作用。拟议的研究将检验TSLP是发展和维持特应性和哮喘的总开关的总体假设。TSLP是一种造血细胞因子，编码在人类染色体5q22的Th2细胞因子基因簇上，在人类中主要由上皮细胞和激活的肥大细胞产生，并在特应性皮炎和过敏性哮喘的背景下表达。过敏性疾病的基因工程小鼠模型也支持TSLP在特应性疾病的病理生物学中的核心作用。该提案包括四个研究项目、两个研究核心和一个侧重于TSLP的行政核心。项目1将研究人类呼吸道上皮细胞和肥大细胞诱导TSLP表达的机制。项目2将确定TSLP激活髓系树突状细胞的机制。项目3将阐明TSLP激活的髓系树突状细胞增殖Th2效应细胞和记忆细胞的机制。项目4将侧重于炎性T辅助细胞(THI)在上调TSLP产生中的作用。这些项目将由一个行政核心、一个分析研究核心和一个临床研究核心提供支持。拟议的研究将为过敏性疾病和哮喘的机制提供新的见解，并可能确定治疗干预的新靶点。 项目1：TSLP表达的调节(休斯顿，D.) 描述(由申请人提供)： 德克萨斯医学中心AADCRC的这项申请是一项跨学科、跨机构、高度集成的计划，重点在于阐明人胸腺基质淋巴生成素(TSLP)在主导过敏表型的Th2免疫反应的发展和维持中的作用。初步数据支持TSLP介导的变态反应性炎症的先天和适应性两阶段模型。提出了四个综合项目和三个核心，以调查TSLP是发展和维持过敏表型的主开关的总体假设。项目1侧重于TSLP表达的调控机制；项目2侧重于髓系树突状细胞(MDC)的TSLP激活；项目3侧重于MDC对Th2免疫的传播；项目4侧重于T细胞对TSLP表达的反馈调节。项目1将测试与特应性和过敏性哮喘有关的空气变应原和呼吸道病毒病原体可以刺激呼吸道上皮细胞和肥大细胞产生TSLP，从而启动过敏性炎症的先天免疫级联反应的假设。目的1探讨呼吸道合胞病毒(RSV)、鼻病毒(RV)和空气变应原在体外诱导人呼吸道上皮细胞和肥大细胞表达TSLP的假设。目的研究TSLP在人呼吸道上皮细胞和肥大细胞中表达上调的假说，利用核因子？B、AP-1和STAT6激活途径。目的研究呼吸道合胞病毒(RSV)、轮状病毒(RV)和空气变应原诱导变应性鼻炎和哮喘患者TSLP表达的体内效应。拟议的研究将利用已有的技术，用于：(I)通过实时荧光聚合酶链式反应、原位杂交、免疫组织学、酶联免疫吸附试验和生物测定测定TSLP；(Ii)培养和分析人类原代支气管和鼻腔上皮细胞、支气管上皮细胞系、CD34来源的肥大细胞和LAD2肥大细胞系；(Iii)通过BCM-NIH病毒呼吸道病原体研究单位提供RSV和RV专业知识；(Iv)使用启动子-荧光素酶结构分析核基因激活；以及(V)对患有和不患有过敏性鼻炎和哮喘的现有患者进行临床研究。研究将利用和利用所有三个核心领域的专业知识。拟议的研究将提供对过敏性炎症的致病机制的洞察，并为开发过敏性哮喘和特应性疾病的创新疗法确定新的靶点。

DESCRIPTION (provided by applicant): This is an application for a Texas Medical Center NIH/NIAID Asthma and Allergic Diseases Cooperative Research Center that unites the University of Texas M.D. Anderson Cancer Center and Baylor College of Medicine to investigate the role of human Thymic Stromal Lymphopoietin (TSLP) in the development and maintenance of the Th2 immune response that dominates the allergic phenotype. The proposed studies will test the overall hypothesis that TSLP is a master switch for developing and maintaining atopy and asthma. TSLP is a hematopoietic cytokine encoded on the human chromosome 5q22 cluster of Th2 cytokine genes, and in humans is produced predominately by epithelial cells and activated mast cells, and is expressed in the context of atopic dermatitis and allergic asthma. Genetically engineered murine models of allergic disease also support a central role for TSLP in the pathobiology of atopy. This proposal integrates four research projects, two research cores and an administrative core focused on TSLP. Project 1 will investigate the mechanisms for induction of TSLP expression by human respiratory epithelial cells and mast cells. Project 2 will determine the mechanisms by which TSLP activates myeloid dendritic cells. Project 3 will elucidate mechanisms by which TSLP activated myeloid dendritic cells propagate Th2 effecter and memory cells. Project 4 will focus on the role of inflammatory T helper cells (Thi) in the upregulation of TSLP production. These projects will be supported by an Administrative Core, an Analytical Research Core, and a Clinical Research Core. The proposed studies should provide new insight into the mechanisms of allergic disease and asthma, and potentially identify new targets for therapeutic intervention. PROJECT 1: Regulation of TSLP Expression (Huston, D.) DESCRIPTION (provided by applicant): This application for a Texas Medical Center AADCRC is an inter-disciplinary, inter-institutional, highly integrated program focused on elucidating the role of human Thymic Stromal Lymphopoietin (TSLP) in the development and maintenance of the Th2 immune response that dominates the allergic phenotype. Preliminary data support an innate and adaptive two-stage model for TSLP-mediated allergic inflammation. Four integrated Projects and three Cores are proposed to investigate the overall hypothesis that TSLP is a master switch for developing and maintaining the allergic phenotype. Project 1 is focused on mechanisms regulating the expression of TSLP; Project 2 is focused on TSLP activation of myeloid dendritic cells (mDC); Project 3 is focused on mDC propagation of Th2 immunity; and Project 4 is focused on T cell feedback regulation on TSLP expression. Project 1 will test the hypothesis that aeroallergens and respiratory viral pathogens linked to atopy and allergic asthma can stimulate respiratory epithelial cells and mast cells to produce TSLP, and thereby initiate the innate immunologic cascade for allergic inflammation. Aim 1 will investigate the hypothesis that respiratory syncytial virus (RSV), rhinovirus (RV), and aeroallergens induce expression of TSLP by human respiratory epithelial cells and mast cells, in vitro. Aim 2 will investigate the hypothesis that upregulation of TSLP expression in human respiratory epithelial cells and mast cells, utilizes NF?B, AP-1, and STAT6 activation pathways. Aim 3 will investigate the in vivo effects of RSV, RV, and aeroallergens to induce expression of TSLP in patients with and without allergic rhinitis and asthma. Proposed studies will utilize established techniques for: (i) measuring TSLP by real-time PCR, in situ hybridization, immunohistology, ELISA, and bioassays; (ii) culture and analysis of human primary bronchial and nasal epithelial cells, bronchial epithelial cell lines, CD34- derived mast cells, and the LAD2 mast cell line; (iii) RSV and RV expertise through the BCM-NIH Viral Respiratory Pathogens Research Unit; (iv) analysis of nuclear gene activation, using promoter-luciferase constructs; and (v) Clinical Research studies with available patients with and without allergic rhinitis and asthma. Studies will utilize and leverage the expertise in all three Cores. The proposed studies will provide insight into the pathogenic mechanisms of allergic inflammation and define novel targets for developing innovative therapies for allergic asthma and atopy.