FLUORESCENCE STUDIES OF T-CELL RECOGNITION

T 细胞识别的荧光研究

• 批准号: 3466590
• 负责人: ADRIENNE A BRIAN
• 金额: $ 9.54万
• 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/3466590
• 关键词: T cell receptor; T lymphocyte; antigen receptors; antireceptor antibody; calcium transporting ATPase; cell population study; chemical binding; clone cells; cytotoxic T lymphocyte; 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细胞中免疫功能的激活需要 抗原的特异性受体和 由主要跨膜糖蛋白编码 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。 也就是说，通过抗体的交联， 不存在抗原，足以刺激功能性 反应 本建议中所描述的实验旨在 确定是否存在显著的受体诱导的 抗原存在于T细胞和靶之间的接触区域中 膜的 靶膜将由平面的 磷脂双分子层支撑在玻璃上， 已经被重组了。 支撑膜以前具有 被证明在刺激免疫功能方面有效 来自MHC限制性抗原特异性T细胞 开始前 检测膜组分的再分布的实验， 将集中精力纠正某些缺点， 支撑膜，特别是从朝向 产生支持膜，其中MHC分子 横向移动的。 为了提高重组效率，我们 我将尝试几种方法将MHC抗原插入到 预成型的支撑双层。 功能测定将用于 确定MHC横向迁移率变化的影响， 分子。 对于识别I类分子的细胞毒性T细胞， 将尝试一些功能性反应，尤其是诱导 这将使研究模型 膜延伸到这个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。 的 再分布的程度应反映受体的亲和力 MHC配体。 这些实验的结果应该 受体参与细胞-细胞 在其他生物和发育系统中的识别。