BRAIN REACTIVE AUTOANTIBODIES--ALZHEIMER'S DISEASE

脑反应性自身抗体--阿尔茨海默病

• 批准号: 3078636
• 负责人: CHRISTINE M HULETTE
• 金额: $ 6.37万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3078636
• 关键词: Alzheimer's disease; aging; antibody specificity; autoantibody; brain; complementary DNA; enzyme linked immunosorbent assay; epitope mapping; genetic library; histopathology; human tissue; immunocytochemistry; lymphoblast; monoclonal antibody; nucleic acid probes; protein purification; tissue /cell culture

Evidence from several investigators suggests that BRA are present in a spectrum of antigenic specificities in Alzheimer's Disease (AD) and provide conceptual support for their pathogenetic role. Endogenous BRA will be demonstrated by direct immunohistochemical staining of AD and control brain tissue from the rapid autopsy protocol (RAP) of Duke University, Serum titers of BRA will be determined by Western blot of human brain homogenate. Regional distribution of autoantigenic specificities will be demonstrated by indirect immunohistochemical staining of RAP brian tissue and normal young adult brain tissue using AD and aged control sera. Previous work by the PI in collaboration with Roy Walford, M.D. has demonstrated that lymphoblastoid cell lines (LCL) from individuals secreting high titer antibodies may be used to created human- human hybridomas secreting antibodies of desired specificity which can be purified and characterized. This technique will be used to develop human monoclonal BRA which can be exploited to characterize the spectrum of antigenic specificities in AD and normal aged by immunoblot, tissue distribution, blocking and cytotoxicity studies. The Alzheimer's Disease research Center at Duke, directed by Allen Roses, M.D., offers a unique resource for investigation of molecular immunological mechanisms in the pathogenesis of AD. The tissue obtained through the RAP is invaluable to this endeavor. In addition, the Center has available LCLs and serum from several pedigrees of late onset AD. This material is the core of Dr. Roses' investigation of the molecular genetics of AD. In order to round out my training during this award period, I will become trained in the molecular genetic techniques in routine use in Dr. Roses' laboratory. My aim will be to use human monoclonal BRA to unique epitopes, for screening of a human brain with cDNA expression library so that purified antigen is available for further studies.

几位研究人员的证据表明，BRA 存在于 阿尔茨海默氏病 (AD) 的抗原特异性谱 为其致病作用提供概念支持。 内源性BRA 将通过 AD 的直接免疫组织化学染色来证明 通过杜克大学的快速尸检方案（RAP）控制脑组织 大学，BRA 血清滴度将通过蛋白质印迹测定 人脑匀浆。 自身抗原的区域分布 特异性将通过间接免疫组织化学来证明 使用 AD 对 RAP 脑组织和正常年轻成人脑组织进行染色 和老化对照血清。 PI 之前与 Roy 合作的工作 医学博士沃尔福德 (Walford) 已证明，淋巴母细胞系 (LCL) 分泌高效价抗体的个体可用于创建人类- 人类杂交瘤可分泌具有所需特异性的抗体 纯化并表征。 该技术将用于开发人类 单克隆 BRA 可用于表征谱 通过免疫印迹、组织检测 AD 和正常老年人的抗原特异性 分布、阻断和细胞毒性研究。 杜克大学阿尔茨海默病研究中心由艾伦·罗斯 (Allen Roses) 领导， 医学博士，为分子研究提供了独特的资源 AD发病机制中的免疫学机制。 获得的组织 通过 RAP 对这一努力具有无价的价值。 此外，该中心 拥有来自多个迟发性 AD 谱系的可用 LCL 和血清。 这种材料是 Roses 博士分子研究的核心 AD 的遗传学。 为了在这个奖励期间完成我的培训，我将成为 在 Roses 博士的诊所接受过日常使用的分子遗传学技术培训 实验室。 我的目标是利用人类单克隆 BRA 来实现独特的 表位，用于用 cDNA 表达文库筛选人脑 纯化的抗原可用于进一步研究。