FLUORESCENCE STUDIES OF T-CELL RECOGNITION

T 细胞识别的荧光研究

• 批准号: 3466589
• 负责人: ADRIENNE A BRIAN
• 金额: $ 8.12万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 1993-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3466589
• 关键词: T cell receptor; T lymphocyte; antibody dependent killer cell; antigen receptors; antireceptor antibody; calcium transporting ATPase; cell population study; chemical binding; clone cells; fluorescence microscopy; histocompatibility antigens; hybridomas; immunoconjugates; interferons; interleukin 2; laboratory mouse; laboratory rat; leukocyte activation /transformation; membrane activity; membrane lipids; membrane model; membrane proteins; membrane reconstitution /synthesis

The activation of immune function in T cells requires the simultaneous binding by a specific receptor of antigen and a transmembrane glycoprotein coded for by the major histocompatibility complex. As with other cell surface receptors, there is indirect evidence that cross-linking of T cell receptors precedes activation. That is, cross-linking by antibody, in the absence of antigen, is sufficient to stimulate the functional response. Experiments are described in this proposal are aimed at determining whether significant receptor-induced accumulation of antigen occurs in the contact region between a T cell and a target membrane. The target membrane will consist of a planar phospholipid bilayer supported on glass into which MHC antigens have been reconstituted. Supported membranes previously have been shown to be effective in the stimulation of immune function from MHC restricted, antigen specific T cells. Before undertaking experiments to detect redistribution of membrane components, effort will be concentrated on correcting certain shortcomings of the supported membranes, particularly with a view toward producing supported membranes in which the MHC molecules are laterally mobile. To increase the efficiency of reconstitution, we shall try several approaches to inserting MHC antigens into preformed supported bilayers. Functional assays will be used to determine the effects of variation in the lateral mobility of MHC molecules. For cytotoxic T cells recognizing class I molecules, a number of functional responses will be tried, especially induction of gamma-interferon, that will allow studies with model membranes to be extended to this T cell subset. Fluorescence microscopy will be used to measure the degree of redistribution of fluorescently tagged, laterally mobile MHC molecules. The degree of redistribution should reflect the affinity of the receptor for the MHC ligand. The results of these experiments should have general implications for receptors participating in cell-cell recognition in other biological and developmental systems.

T细胞免疫功能的激活需要 抗原和抗原的特定受体同时结合 主要编码的跨膜糖蛋白 组织相容性复合体。 与其他细胞表面受体一样， 有间接证据表明 T 细胞受体的交联 在激活之前。 即，通过抗体进行交联， 缺乏抗原，足以刺激功能 回复。 本提案中描述的实验旨在 确定是否有显着的受体诱导积累 抗原出现在 T 细胞和靶标之间的接触区域 膜。 目标膜将由平面组成 玻璃上支持的磷脂双层，其中 MHC 抗原 已被重组。 以前支持的膜有 已被证明可有效刺激免疫功能 来自 MHC 限制性抗原特异性 T 细胞。 进行之前 检测膜成分重新分布的实验， 将着力纠正一些缺点 支撑膜，特别是着眼于 生产支持膜，其中 MHC 分子 横向移动。 为了提高重构效率，我们 应尝试多种方法将 MHC 抗原插入 预制支撑双层。 功能测定将用于 确定 MHC 横向流动性变化的影响 分子。 对于识别 I 类分子的细胞毒性 T 细胞， 将尝试多种功能反应，尤其是诱导 γ-干扰素，这将允许模型研究 膜延伸至该 T 细胞亚群。 荧光 显微镜将用于测量重新分布的程度 荧光标记的、横向移动的 MHC 分子。 这 再分布的程度应反映受体的亲和力 为 MHC 配体。 这些实验的结果应该是 对参与细胞间受体的一般意义 其他生物和发育系统中的识别。