B LYMPHOCYTE TOLERANCE IN HEALTH AND AUTOIMMUNITY

健康和自身免疫中的 B 淋巴细胞耐受

• 批准号: 2182765
• 负责人: DAVID NEMAZEE
• 金额: $ 14.15万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2182765
• 关键词: B lymphocyte; apoptosis; autoantigens; autoimmunity; gene rearrangement; genetic mapping; genetically modified animals; immune tolerance /unresponsiveness; immunoglobulin genes; laboratory mouse; leukocyte activation /transformation; polymerase chain reaction; tissue /cell culture

The applicant seeks a research project grant (RO1) to elucidate the mechanisms of B lymphocyte tolerance. The long-term goal of the project is to understand how autospecific B-cells are controlled, to determine if autoimmune-prone mouse strains have intrinsic B-cell tolerance defects, and, if so, to localize the genes involved. As a basis for this study, mice transgenic for functionally rearranged anti-H-2Kk IgM/IgD antibody genes have been generated. In these mice B- cells are tolerized in the presence of the appropriate class I antigens. The Specific Aims of this proposal are as follows: 1. To determine the mechanisms of peripheral tolerance in autospecific B-cells. We will follow up the results we obtained in our initial grant with the analysis of tolerance to peripherally-expressed antigen. To do this we will analyze tolerance to membrane autoantigen that is not expressed in the bone marrow, where B-cells develop in the adult mouse, but on the surfaces of cells found in other organs, such as the liver, skin and pancreas. In this analysis we will take advantage of existing transgenic mice, produced by others, that target the expression of the low affinity ligand Kb to these peripheral tissues. This approach works because the antibody encoded by the transgenes has a low, but physiologically significant affinity for the Kb antigen. 2. To determine the influence of autoimmune-prone genetic background on self-tolerance to peripheral membrane antigen. We have already established that in at least one transgenic line expressing Kb in the periphery, autospecific anti-H-2K B-cells are tolerized by deletion. We will cross these transgenes onto autoimmune-prone strains of mice to determine if they manifest a B-cell tolerance defect to Kb antigen in this double transgenic system. 3. To begin the chromosomal mapping of a background gene of MRL strain mice that appears to favor the breaking of self-tolerance in the B-cells of MRL/lpr/3-83 transgenic backcross mice. We will use the most modern technique in classical genetic mapping, including taking advantage of polymorphic DNA microsatellite polymerase chain reaction markers. In this process we will determine if the transgenic system provides advantages of speed and accuracy to the mapping of genes that affect autoimmunity.

申请人寻求研究项目赠款（RO1）来阐明 B淋巴细胞耐受性的机制。 该项目的长期目标 是了解如何控制自动B细胞，以确定 如果自身免疫性易免疫的小鼠菌株具有内在的B细胞耐受性 缺陷，如果是这样，可以定位所涉及的基因。 作为这项研究的基础，小鼠在功能重新排列的转基因 抗H-2KK IgM/IgD抗体基因已产生。 在这些小鼠中b- 在适当的I类抗原存在下耐受细胞。 该提案的具体目的如下： 1。确定自动特异性外周耐受性的机制 B细胞。 我们将跟进我们在初始赠款中获得的结果 通过分析对周围表达的抗原的耐受性。 做 我们将分析对膜自动抗原的耐受性 在骨髓中表达，在成年小鼠中出现B细胞， 但是在其他器官（例如肝脏）中发现的细胞表面上， 皮肤和胰腺。 在此分析中，我们将利用现有 由他人产生的转基因小鼠，其靶向表达 低亲和力KB对这些外围组织。 这种方法有效 因为转基因编码的抗体具有低，但 生理上对KB抗原具有显着的亲和力。 2。确定自身免疫性遗传背景对 对周围膜抗原的自耐耐受性。 我们已经 确定在至少一条转基因线中表达Kb 外围，自动抗H-2K B细胞通过缺失耐受。 我们 将这些转基因将小鼠自身免疫性菌株交叉至 确定它们是否表现出B细胞公差缺陷对Kb抗原的缺陷 这个双转基因系统。 3。开始MRL菌株背景基因的染色体映射 似乎有利于B细胞中自我耐受性破坏的小鼠 MRL/LPR/3-83转基因后卷小鼠 我们将使用最现代的 经典遗传图中的技术，包括利用 多态性DNA微卫星聚合酶链反应标记。 在 这个过程我们将确定转基因系统是否提供 速度和准确性的优势，影响影响的基因 自身免疫性。