GUT-DERIVED IMMUNE EFFECTOR CELLS

肠道来源的免疫效应细胞

• 批准号: 2443979
• 负责人: JOHN R KLEIN
• 金额: $ 18.26万
• 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 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2443979
• 关键词: CD28 molecule; T lymphocyte; bone marrow; cell population study; flow cytometry; gut associated lymphoid tissue; hematopoietic stem cells; immunocytochemistry; laboratory mouse; leukocyte activation /transformation; leukopoiesis; monoclonal antibody; tissue mosaicism

The project is a continuation of studies into the immunobiology of murine intestinal intraepithelial lymphocytes (IEL) as a model for understanding human intestinal T cells. The specific aims constitute three interrelated areas of study that will provide an integrated and comprehensive analyses of IEL development, phenotype, and function. Experiments have been designed to identify and characterize precise extrathymic pathway(s) of IEL development from bone marrow stem cells to mature IEL stage. Using methodologies of proven effectiveness, bone marrow-derived IEL precursor cells will be characterized by flow cytometric (FCM) analyses at precise stages during the early phase of IEL reconstitution in euthymic radiation chimeras, and in radiation chimeras constructed from thymectomized mice and congenitally-athymic nude mice. Specific bone marrow stem cells capable of IEL repopulation will be determined. The precise locations of extrathymic development within the small intestine will be identified by immunoperoxidase staining of intestine tissues. Changes in the IEL T cell repertoire will be identified by immunoperoxidase staining of intestine tissues. Changes in the IEL T cell repertoire will be evaluated during development and throughout life. The thymopoietic nature of the murine small intestine will be studied in mice engrafted with sterile intestine, and the possibility of a resident intestinal pluripotent stem cell will be explored. A second series of experiments will characterize adult mouse IEL phenotypically by multi-color FCM analyses using markers not yet applied to IEL studies to correlate IEL developmental lineages with defined IEL subsets. Newly identified and/or novel IEL subsets will be studied by extensive FCM analyses to gain insight into the functional involvement of those cells. Age-related and genetically-determined changes in IEL phenotypes will be studied. IEL-specific monoclonal antibodies recently isolated in this laboratory will be used in studies of IEL, and additional monoclonal antibodies will be generated to identify new IEL- specific markers. CD28 expression will be studied for IEL and other lymphoid tissues, and the molecular heterogeneity of CD28 within the murine immune system will be explored. A third series of experiments will examine functional properties of murine IEL correlated with information from studies of IEL development and phenotype. Proliferation and cytotoxic properties will be studied in phenotypically-defined, Percoll-fractionated IEL subsets and according to states of activation. The functional role of TCR-gammadelta+ IEL expressing Vgamma5 and Vgamma1.2 will be studied using anti-TCR clonotypic antibodies, and antigen recognition by IEL will be explored using recently isolated IEL-derived T cells clones with known reactivities. Collectively, these studies will provide basic information about IEL development, phenotype, and immune function in order to ultimately understand human IEL in health and disease states.

该项目是对小鼠免疫生物学研究的延续， 肠上皮内淋巴细胞（IEL）作为理解模型 人类肠道T细胞 具体目标包括三个方面 相互关联的研究领域，将提供一个综合的， IEL发育、表型和功能的综合分析。 实验已经被设计来识别和表征精确的 IEL从骨髓干细胞发育为 成熟IEL期。 使用有效的方法，骨 骨髓来源的IEL前体细胞将通过流动来表征 在IEL早期的精确阶段进行流式细胞术（FCM）分析 正常胸腺辐射嵌合体和辐射嵌合体中的重建 由切除胸腺的小鼠和先天无胸腺的裸鼠构建。 能够IEL再增殖的特异性骨髓干细胞将被 测定 胸腺外发育的精确位置 小肠将通过免疫过氧化物酶染色进行鉴定， 肠组织 IEL T细胞库的变化将是 通过肠组织的免疫过氧化物酶染色鉴定。 变化 在IEL T细胞库中，将在发育期间进行评价， 在生活中。 小鼠小肠的胸腺生成特性 将在植入无菌肠的小鼠中进行研究， 肠道多能干细胞的可能性将是 探讨了 第二系列实验将描述成年小鼠IEL的特征。 使用尚未应用的标记物通过多色FCM分析进行表型分析 IEL研究，将IEL发育谱系与定义的IEL相关联 子集 新发现的和/或新的IEL子集将由以下研究人员进行研究： 广泛的FCM分析，以深入了解功能参与 这些细胞。 IEL中的遗传相关和遗传决定的变化 将研究表型。 IEL特异性单克隆抗体最近 本实验室分离的细菌将用于IEL研究， 将产生额外的单克隆抗体来鉴定新的IEL， 特定的标记。 将研究CD 28表达的IEL和其他 淋巴组织中CD 28的分子异质性， 将探索鼠免疫系统。 第三系列实验将检查 小鼠IEL与来自IEL发展研究的信息相关 和表型。 将研究增殖和细胞毒性特性 在表型定义的Percoll分级IEL子集中， 到激活状态。 TCR-γ δ + IEL的功能作用 将使用抗TCR研究表达V γ 5和V γ 1.2的细胞 克隆型抗体和抗原识别IEL将进行探讨 使用最近分离的IEL衍生的T细胞克隆， 反应性 总的来说，这些研究将提供基本信息 了解IEL的发育、表型和免疫功能， 最终了解健康和疾病状态下的人类IEL。