CLASS II GENES IN AUTOIMMUNE RODENTS

自身免疫啮齿动物中的 II 类基因

• 批准号: 3141525
• 负责人: JOEL SCHIFFENBAUER
• 金额: $ 15.34万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3141525
• 关键词: DNA; MHC class II antigen; autoimmune disorder; electrophoresis; genetic library; genetic manipulation; genetic regulatory element; genetically modified animals; laboratory mouse; monoclonal antibody; nucleic acid hybridization; protein sequence; protein structure function; transcription factor; transfection; western blottings

The class II molecules play a central role in both normal and abnormal immune responses, and a number of autoimmune diseases both in man and mouse appear to be linked to class II molecules. We propose to evaluate the qualitative and quantitative contribution of the class II molecules from the NZW mouse strain to autoimmunity in the (NZWxNZB)F mouse. The qualitative structure function relationship of class II molecules in autoimmunity will be evaluated by seeking unusual sequences in NZW cDNA's. We will develop monoclonal antibodies specific for the NZW/NZB hybrid class II molecules to examine whether hybrid class II molecules exist in vivo, and determine if these hybrid molecules might contribute to the development of autoimmunity. Using transfections we will also determine whether the "u" haplotype is immunodominant, which would help explain the NZW contribution to accelerated autoimmunity. In addition, we plan to produce transgenic NZB mice, in which NZW class II transgenes are placed into NZB mice. To evaluate whether there is a quantitative contribution of class II molecules to autoimmunity, we will isolate NZB and NZW genes and examine both cis and trans acting factors which regulate the expression of class II genes. Cis acting factors will be evaluated by determining whether the 5' flanking region of the class II genes of NZW and NZB have unusual sequences. cDNA's encoding trans acting factors will be isolated by a Southwestern methodology whereby labeled oligonucleotides corresponding to the NZB and NZW 5' regulatory regions are used to probe NZB and NZW lambda gt11 libraries. These cDNA's will be examined for unusual sequences. These studies will evaluate both the quantitative and qualitative contributions of class II genes to autoimmunity.

II类分子在正常和异常中都发挥着核心作用 免疫反应以及人类和人类的许多自身免疫性疾病 小鼠似乎与 II 类分子有关。 我们建议评估 II 类分子的定性和定量贡献 从 NZW 小鼠品系到 (NZWxNZB)F 小鼠的自身免疫。 这 II类分子的定性结构功能关系 将通过寻找 NZW cDNA 中的异常序列来评估自身免疫。 我们将开发针对 NZW/NZB 的单克隆抗体 杂合 II 类分子，检查是否杂合 II 类分子 存在于体内，并确定这些杂合分子是否可能有助于 自身免疫的发展。 使用转染，我们还将 确定“u”单倍型是否是免疫显性的，这将有助于 解释 NZW 对加速自身免疫的贡献。 此外，我们 计划生产转基因NZB小鼠，其中NZW II类转基因 放入NZB小鼠体内。 评价是否有II类定量贡献 分子对自身免疫的影响，我们将分离 NZB 和 NZW 基因并检查 调节类表达的顺式和反式作用因子 二基因。 顺式作用因素将通过确定是否 NZW和NZB II类基因的5'侧翼区域有异常 序列。 编码反式作用因子的 cDNA 将通过 西南方法论，标记的寡核苷酸对应于 NZB 和 NZW 5' 调控区用于探测 NZB 和 NZW lambda gt11 库。 将检查这些 cDNA 是否存在异常序列。 这些研究将评估定量和定性 II类基因对自身免疫的贡献。