ANTI-SM B CELLS OF MRL/LPR MICE

MRL/LPR 小鼠的抗 SM B 细胞

• 批准号: 3748043
• 负责人: Stephen H Clarke
• 金额: --
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3748043
• 关键词: B lymphocyte; DNA binding protein; antibody formation; antibody specificity; autoantibody; cellular pathology; gene mutation; genetic strain; genetically modified animals; immunoregulation; laboratory mouse; leukocyte activation /transformation; molecular cloning; molecular pathology; polymerase chain reaction; ribonucleoproteins; systemic lupus erythematosus

The long term objective of this proposal is to understand the events in B cell development and selection that lead to the production of autoantibodies in MRL/Mp-lpr/lpr (MRL/lpr) mice. Mice of this strain develop a spontaneous autoimmune disease that resembles systemic lupus erythematosus (SLE). We have begun a study of the B cell response to the Sm particle, a ribonucleoprotein present in the nuclei of all cells. The spontaneous response to this particle in humans is diagnostic of SLE, and MRL/lpr mice are the only mouse model that spontaneously develops a response to this antigen. The correlation of the response to Sm and SLE suggests an essential relationship between the etiology of the disease and the production of these autoantibodies. Our previous analysis indicates that Sm-specific B cells are selected by DNA, but also indicates the involvement of a second antigen, presumably Sm. We propose in Aim 1 to test the hypothesis that Sm is a selecting antigen in this response. This will be accomplished by identifying the mutations in multiple anti-Sm hybridomas, and determining whether their distribution is biased, an indication of antigen selection of mutant B cells. In addition, through the use of transfectoma antibodies we will determine whether the observed mutations improve Sm and DNA binding. In Aim 2 we will examine the basis for the dual Sm and DNA binding of anti-Sm selected hybridomas. We propose that DNA binding is determined principally by the H chain and that Sm binding is determined principally by the L chain. This hypothesis will be tested by measuring Sm and DNA binding of generated transfectomas antibodies that differ in the VH or Vk. In Aim 3 we will generate transgenic mice using VH and Vk genes of anti-Sm and anti-Sm/DNA hybridomas. Transgenic mice will be crossed onto both normal and autoimmune genetic backgrounds to examine the immunoregulation of these cells in normal mice and their disregulation in autoimmune mice.

该提案的长期目标是了解 B 中的事件 细胞的发育和选择导致产生 MRL/Mp-lpr/lpr (MRL/lpr) 小鼠中的自身抗体。该品系小鼠 出现类似于系统性狼疮的自发性自身免疫性疾病 红斑狼疮（SLE）。我们已经开始研究 B 细胞对 Sm颗粒，一种存在于所有细胞核中的核糖核蛋白。这 人类对这种粒子的自发反应可以诊断 SLE，并且 MRL/lpr 小鼠是唯一能够自发形成 对这种抗原的反应。 Sm 反应与 SLE 的相关性 表明该疾病的病因学与 这些自身抗体的产生。我们之前的分析表明 Sm 特异性 B 细胞是由 DNA 选择的，但也表明 涉及第二种抗原，大概是Sm。我们在目标 1 中建议 检验 Sm 是该反应中的选择抗原的假设。这 将通过鉴定多个抗Sm中的突变来完成 杂交瘤，并确定它们的分布是否有偏差， 突变 B 细胞抗原选择的指示。此外，通过 使用转染瘤抗体，我们将确定是否观察到 突变改善了 Sm 和 DNA 的结合。在目标 2 中，我们将检查基础 用于抗 Sm 选择的杂交瘤的 Sm 和 DNA 双重结合。我们建议 DNA 结合主要由 H 链决定，Sm 结合主要由L链决定。这个假设将是 通过测量生成的转染瘤的 Sm 和 DNA 结合进行测试 VH 或 Vk 不同的抗体。在目标 3 中，我们将生成 使用抗Sm和抗Sm/DNA的VH和Vk基因的转基因小鼠 杂交瘤。 转基因小鼠将与正常小鼠和正常小鼠杂交 自身免疫遗传背景来检查这些的免疫调节 正常小鼠的细胞及其在自身免疫小鼠中的失调。