GUT-DERIVED IMMUNE EFFECTOR CELLS

肠道来源的免疫效应细胞

• 批准号: 2139600
• 负责人: JOHN R KLEIN
• 金额: $ 16.59万
• 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/2139600
• 关键词: GUT; DERIVED; IMMUNE; EFFECTOR; CELLS

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继续。 肠道上皮内淋巴细胞(IEL)作为一种理解模型 人体肠道T细胞。具体目标包括三个方面 相互关联的研究领域，将提供一个综合和 IEL的发育、表型和功能的综合分析。 已经设计了实验来识别和表征精确的 骨髓干细胞向胸腺外发育的胸腺外途径(S) 成熟的IEL阶段。使用已被证明有效的方法，伯恩 骨髓来源的IEL前体细胞将以Flow为特征 IEL早期精确分期的流式细胞仪分析 在胸腺辐射嵌合体和辐射嵌合体中的重建 由胸腺切除小鼠和先天性裸鼠构建而成。 具有IEL再分化能力的特定骨髓干细胞将被 下定决心。胸腺外发育的准确位置 小肠将通过免疫过氧化物酶染色来识别 肠道组织。IEL T细胞谱系的变化将是 通过肠道组织免疫过氧化物酶染色鉴定。变化 在IEL T细胞谱系中，将在发育和 在一生中。小鼠小肠的胸腺生成功能 将在植入无菌肠道的小鼠身上进行研究， 移植肠道多能干细胞的可能性将是 探索过了。 第二系列实验将描述成年小鼠的IEL 使用尚未应用的标记进行多色FCM分析的表型 对IEL研究将IEL发育谱系与已定义的IEL联系起来 子集。将通过以下方式研究新识别的和/或新的IEL子集 广泛的FCM分析以深入了解功能参与 这些细胞。IEL的年龄相关和遗传决定的变化 将对表型进行研究。IEL特异性单抗的研究进展 在这个实验室中分离出来的将用于IEL的研究，以及 将产生更多的单抗来鉴定新的IEL- 特定的标记。将对IEL和其他疾病的CD28表达进行研究 淋巴组织中CD28的分子异质性。 将探索小鼠的免疫系统。 第三个系列的实验将检验 小鼠IEL与IEL发育研究中的信息相关 和表型。将研究细胞增殖和细胞毒特性。 在表型定义的Percoll分步IEL子集中 变为激活状态。TCR-Gammadelta+IEL的功能作用 表达VGamma5和VGamma1.2将使用抗TCR进行研究 将探索克隆型抗体和IEL对抗原的识别 使用最近分离的IEL来源的T细胞克隆 反应性。总的来说，这些研究将提供基本信息 关于IEL的发育、表型和免疫功能 最终了解人类IEL在健康和疾病状态下的情况。