Antigen Presentation in Human Autoimmune Diseases

人类自身免疫性疾病中的抗原呈递

• 批准号: 7027086
• 负责人: Kai W Wucherpfennig
• 金额: $ 82.09万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7027086
• 关键词: antigen presentation; autoimmune disorder; clinical research; multiple sclerosis

DESCRIPTION (provided by applicant): This program project will continue to focus on antigen presentation and T cell recognition in patients with multiple sclerosis. During the past three years considerable efforts were directed at establishing and validating new technology, now in place for the continuation of the PPG. These novel approaches will be used to visualize antigen presentation in the central nervous system and to molecularly define the T cell and B cell populations involved in the disease process. Project 1 has developed a novel approach for the generation of MAC class II terriers, which will be used to quantify and characterize antigen-specific COD T cells in MS patients and a humanized mouse model of the disease. Project 2 has isolated expanded populations of T cells from the cerebrospinal fluid of patients with MS and will characterize the antigen receptors of T cell and B cell populations. Combinatorial peptide libraries and MHC class II terriers will be used to define the antigen specificity of T cell clones established from the cerebrospinal fluid. B cells will be isolated from cerebrospinal fluid by FACES and MS plaque tissue by laser capture micro dissection, so that recombinant antibodies can be expressed based on the isolated heavy and light chain sequences. Such recombinant antibodies provide an abundant source of these proteins for definition of their target specificity, to be established by a polemics approach, and assessment of pathogenicity. Project 3 will examine the entry of antigen presenting cells into the CNS using a newly created mouse model in which a GFP moiety is attached to the MHC class II beta chain through knock-in technology. The interaction of these GFP expressing antigen presenting cells with infiltrating T cells will be investigated in the optic nerve of live animals by 2-photon microscopy. Each of the investigators contributes a unique area of expertise and innovative approaches to this important clinical problem. The resulting program is highly synergistic and will allow the application of powerful new tools in proteomics and biochemistry to the pathophysiology of human inflammatory disease.

描述（由申请人提供）：该计划项目将继续关注多发性硬化症患者的抗原呈递和T细胞识别。 在过去三年中，为建立和验证新技术做出了相当大的努力，现在已经到位，以继续PPG。 这些新的方法将被用于可视化中枢神经系统中的抗原呈递，并从分子上定义参与疾病过程的T细胞和B细胞群体。 项目1开发了一种新的方法，用于产生MAC II类梗犬，这将被用来量化和表征抗原特异性COD T细胞在MS患者和人源化小鼠模型的疾病。 项目2已从MS患者的脑脊液中分离出扩增的T细胞群，并将表征T细胞和B细胞群的抗原受体。 组合肽库和MHC II类梗类将用于定义从脑脊液建立的T细胞克隆的抗原特异性。 将通过FACES从脑脊液中分离B细胞，并通过激光捕获显微切割从MS斑块组织中分离B细胞，从而可以基于分离的重链和轻链序列表达重组抗体。 这种重组抗体提供了这些蛋白质的丰富来源，用于通过争论方法确定其靶特异性和评估致病性。 项目3将使用新创建的小鼠模型检查抗原呈递细胞进入CNS，在该模型中，GFP部分通过敲入技术连接到MHC II类β链。 将通过双光子显微镜在活体动物的视神经中研究这些表达GFP的抗原呈递细胞与浸润性T细胞的相互作用。 每个研究人员都为这一重要的临床问题贡献了独特的专业知识和创新方法。 由此产生的程序是高度协同的，并将允许在蛋白质组学和生物化学的人类炎症性疾病的病理生理学的强大的新工具的应用。