ANTIGEN PRESENTATION AND SELF TOLERANCE

抗原呈递和自我耐受

• 批准号: 3085350
• 负责人: DAVID T HAGERTY
• 金额: $ 7.29万
• 依托单位: JEWISH HOSPITAL OF SAINT LOUIS
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3085350
• 关键词: T lymphocyte; antigen presentation; antigen presenting cell; clone cells; hemoglobin; histocompatibility; histocompatibility antigens; immune tolerance /unresponsiveness; laboratory mouse; leukocyte activation /transformation; stress proteins; tissue /cell culture

The purpose of this proposal is to solicit support through the Physician Scientist Award for studies examining the relationship of the handling of self proteins by antigen presenting cells to self tolerance and presentation of foreign antigens. The results will then be applied to the study of alloreactivity with a long term goal of using the knowledge and insights from our studies to find a method of inducing specific unresponsiveness in an allograft model. The hallmark of the immune response is the ability to distinguish self from nonself. However, the mechanisms by which the immune system develops and maintains tolerance to self antigens, while maintaining reactivity toward foreign antigens, are poorly understood. On of the mechanisms of self tolerance appears to involve the deletion of self-reactive cells in the thymus during T cell maturation. However, this mechanism alone does not completely explain self tolerance. Self proteins, like foreign proteins, have been shown in vitro to be processed and presented by antigen presenting cells. Until recently, however, it was not known whether this phenomenon actually occurred in vivo. Lorenz and Allen have described a unique system for studying the in vivo presentation of the self hemoglobin molecule. Using this system, they were able to show not only that self macrophages were able to present self hemoglobin molecules, but that a variety of cells in different organs of the body had self hemoglobin/Ia molecules on their surfaces that could be recognized by appropriate T cell hybridomas. As the mice do not react to these self hemoglobin/Ia molecules in vivo, T cell tolerance has occurred. Research proposal. For the first two years of the Award, David Hagerty will work in the group of Emil Unanue in the laboratory of Paul M. Allen. He will learn the laboratory techniques of creating and maintaining T cell hybridomas, cell and tissue culture, sterile technique, antigen presentation assays, cell purification and how to set-up RIA and ELISA assays. During this time he proposes to study in more detail the processing and presentation of self proteins by macrophages and other antigen representing cells in an attempt to understand the relationship between the processing and presentation of self and foreign proteins and self tolerance. Using the murine T cell hybridoma YO1.6, the I-Ek restricted hybrid which detects the processed hemoglobin peptide fragment HbBdminor(67-76), he will isolate and purify different cells in each organ homogenate and determine which cells are expressing the self Hb/Ia complexes. He will then use self-Hb specific TH1 and TH2 T cell clones to study the ability of the identified APCs from the organ homogenates to stimulate the clones, especially with regard to any costimulators that may be required. He will then study the mechanisms of tolerance to other self antigens that are compartmentalized and tissue specific (GFAP) or developmentally regulated (sperm hsp70). He plans to apply the findings of the above experiments to the study of alloreactivity, and eventually to apply these results to the achievement of specific unresponsiveness in an organ transplant model.

该提议的目的是通过医生征求支持 研究研究的研究奖 通过抗原呈现细胞自耐力的自我蛋白质和 外国抗原的表现。 然后将结果应用于 使用知识和 我们研究的见解，以找到一种诱导特定的方法 在同种异体移植模型中无反应。 免疫反应的标志是区分自我的能力 来自非自然。 但是，免疫系统发展的机制 并保持对自抗原的耐受性，同时保持反应性 迈向外国抗原，知之甚少。 关于机制 自耐力似乎涉及自反应细胞的缺失 T细胞成熟过程中的胸腺。 但是，仅这种机制就可以 不能完全解释自负。 自我蛋白质，例如异物蛋白，已在体外显示为 通过抗原呈递细胞进行处理和提出。 直到最近， 但是，尚不清楚这种现象是否真的发生在 体内。 洛伦兹（Lorenz）和艾伦（Allen）描述了一个独特的系统来研究 自我血红蛋白分子的体内表现。 使用此系统， 他们不仅能够表明自我巨噬细胞能够表现出来 自我血红蛋白分子，但不同器官中的各种细胞 身体有自我血红蛋白/ia 表面上的分子可以通过适当的T细胞识别 杂交瘤。 由于小鼠对这些自我血红蛋白/IA不反应 分子在体内，T细胞耐受性发生。 研究建议。 奖项的头两年，大卫·哈格蒂（David Hagerty） 将在保罗·M·艾伦（Paul M. Allen）实验室中的埃米尔（Emil）Unanue小组中工作。 他将学习创建和维护T细胞的实验室技术 杂交瘤，细胞和组织培养，无菌技术，抗原 演示分析，细胞净化以及如何设置RIA和ELISA 测定。 在此期间，他建议更详细地研究 自我的处理和表现 巨噬细胞和其他代表细胞的蛋白质蛋白质尝试 了解处理和表现之间的关系 自我和外国蛋白质和自耐受性。 使用鼠T细胞 杂交瘤YO1.6，I-EK限制的混合体，检测到已处理的 血红蛋白肽片段hbbdminor（67-76），他将分离并纯化 每个器官匀浆中的不同细胞并确定哪些细胞是 表达自我HB/IA复合体。 然后，他将使用自我HB特定 Th1和Th2 T细胞克隆研究已鉴定的APC的能力 器官匀浆以刺激克隆，尤其是 任何可能需要的costimulators。 然后他将研究机制 对隔室和组织的其他自抗原的耐受性 特定（GFAP）或发育调节（精子HSP70）。 他计划 将上述实验的发现应用于研究 同种异体性，并最终将这些结果应用于成就 器官移植模型中特定的无反应性。