GUT DERIVED IMMUNE EFFECTOR CELLS

肠道来源的免疫效应细胞

• 批准号: 2905328
• 负责人: JOHN R KLEIN
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF TULSA
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2905328
• 关键词: RNase protection assay; T cell receptor; T lymphocyte; autoimmune disorder; cell differentiation; cytokine; developmental immunology; enteritis; enzyme linked immunosorbent assay; gastrointestinal epithelium; genetic regulatory element; genetically modified animals; gut associated lymphoid tissue; hormone receptor; hormone regulation /control mechanism; immunoprecipitation; immunoregulation; laboratory mouse; leukocyte activation /transformation; mucosal immunity; passive immunization; polymerase chain reaction; thyrotropin

This project is a continuation of studies into the immunobiology of murine intestinal intraepithelial lymphocytes (IELs) as a model for understanding human intestinal T cells in health and disease. The studies described here are derived from work done during the previous funding period, and are designed to address specific unresolved aspects of intestinal T cell development, and to obtain detailed information into intestinal immune-endocrine interactions in the regulation of local intestinal immunity and gastrointestinal disease. The goals of this project are: Specific Aim 1 - to characterize regional sites of local intestinal IEL maturation and development; to determine the lineage relationships between IEL precursors and mature intestinal T cells using intestine- and bone marrow-derived precursors; to determine the potential contribution of mature peripheral T cells and immature thymocytes to the overall intestinal T cell pool. These studies will use transgenic mice which express high levels of a green fluorescent protein in adoptive transfer experiments to trace IEL developmental pathways through phenotypic studies, and by molecular analyses of IEL Vgamma,delta, and beta TCR repertoires. Specific Aim 2 - to systematically characterize the intestinal thyrotropin (thyroid- stimulating hormone [TSH])-mediated immune-endocrine network in terms of local hormone production and utilization in phenotypically-defined cell populations; to study their immunoregulatory effects on intestinal IELs; to delineate biochemical mechanisms of TSH-mediated intestinal T cell activation. RT-PCR analyses of hormone receptor gene expression and hormone binding assays will be used to accurately delineate pathways of intercellular communication; RNase protection assay, ELISA, and ELISPOT assays will be used to identify cytokine and inflammatory mediators activated by TSH; and immunoprecipitation of phosphorylated Jak/STAT regulatory elements will be done to identify TSH receptor- mediated IEL activation. Specific Aim 3 -to further explore the capacity of hypothalamus-pituitary-thyroid hormones to influence or alter gastrointestinal autoimmunity and inflammation using established animal models of those diseases. Phenotypic analyses will be done using lymphocytes from gastrointestinal inflammatory sites, and characterization of hormone-activated regulatory and effector cells will be identified through adoptive transfer experiments.

该项目是对免疫生物学研究的延续， 小鼠肠上皮内淋巴细胞（IEL）作为模型， 了解健康和疾病中的人类肠道T细胞。 的 这里描述的研究来自以前的工作， 供资期间，并旨在解决具体的未决问题 肠道T细胞发育，并获得详细信息 肠道免疫内分泌相互作用的调节， 肠道免疫和胃肠道疾病。 这个的目标 具体目标1 -确定当地 肠IEL成熟和发育;以确定谱系 IEL前体和成熟肠T细胞之间的关系， 肠和骨髓来源的前体;以确定 成熟外周T细胞和未成熟T细胞的潜在贡献 胸腺细胞与整个肠道T细胞池的比例。 这些研究将 使用表达高水平绿色荧光的转基因小鼠， 蛋白质在过继转移实验中追踪IEL发育 通过表型研究和IEL的分子分析， V γ、δ和β TCR库。 具体目标2至 系统地表征肠促甲状腺激素（甲状腺- 促甲状腺激素（TSH）介导的免疫-内分泌网络 局部激素的产生和利用， 细胞群;研究其对肠道免疫调节作用， 描述促甲状腺激素介导的肠T细胞增殖的生化机制 细胞激活 激素受体基因表达的RT-PCR分析 激素结合分析将被用来准确地描绘途径 RNA酶保护试验，ELISA，和 ELISPOT测定将用于鉴定细胞因子和炎性细胞因子。 促甲状腺激素激活的介质;和磷酸化的免疫沉淀 将进行Jak/STAT调节元件以鉴定TSH受体- 介导的IEL激活。 具体目标3 -进一步探讨 下丘脑-垂体-甲状腺激素影响或 改变胃肠道自身免疫和炎症， 这些疾病的动物模型。表型分析将使用 来自胃肠道炎症部位的淋巴细胞，和 对转录激活的调节细胞和效应细胞的表征将 通过过继转移实验来鉴定。