IDENTIFICATION OF AUTOIMMUNE GENES IN LPR/LPR MICE

LPR/LPR 小鼠自身免疫基因的鉴定

• 批准号: 2080451
• 负责人: Michael F. Seldin
• 金额: $ 20.71万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2080451
• 关键词: artificial chromosomes; autoantibody; autoimmune disorder; autoimmunity; disease /disorder model; genetic mapping; genetic markers; genetic polymorphism; glomerulonephritis; human subject; laboratory mouse; molecular cloning; phenotype; restriction mapping; southern blotting

Genetic factors contribute to the expression of autoimmune diseases in both mice and humans. These genes are largely uncharacterized; moreover, their gene products and mechanisms of action are unknown. A recessive mutation, lpr, that occurred in MRL mice results in profound lymphadenopathy with expansion of an unusual subpopulation of T cells and generalized autoimmune disease. A "reverse" genetic approach is proposed to identify the mutant or deficient gene products of the lpr locus and MRL genes that interact with lpr to result in renal disease. The method relies on defining the physical location of these genes. RFLV and microsatellite polymorphisms will be used as markers in two different crosses segregating for the recessive genes that are critical to clinical and serologic disease in these mice. The renal histology will be scored and correlated with the genotype analyses and autoantibody profiles. The localization of lpr to mouse Chr 19 will be confirmed and saturation mapping using clones from a flow sorted mouse Chr 19 library will be performed. Of most interest will be defining the MRL quantitative trait loci (QTL) that predispose to nephritis. Detailed mapping of each relevant region of the genome will be undertaken using available clones localized to the specific chromosome segment and clones likely to be in that region of the genome. If necessary, a subchromosome specific library or a chromosome microdissection library will be generated to allow saturation mapping of one of the identified QTL regions. Long range restriction site analyses of the relevant regions of the mouse genome will be performed. If, as is likely, one or more autoimmune loci are within a chromosomal segment largely conserved between the mouse and human genome, these studies may identify the chromosomal location of human "autoimmune genes". A variety of strategies including the use of chromosome jumping and a mouse yeast artificial chromosome library, in addition to long range restriction mapping will be used to more precisely define the molecular location of the critical gene (s) responsible for genetic predisposition. Comparison with MRL-plus-minus/plus-minus mice will be useful for determining the lpr mutation as will comparison of CBA/KJ with a new mutation at the lpr locus in a strain designated CBA/KJ-lpr(cg) . For other localized "autoimmune genes", candidate genes may be suggested by their chromosomal location. Definition of a single gene that predisposes to autoimmunity should allow fundamental insight into molecular basis for disease in mice and humans.

遗传因素导致自身免疫性疾病的发生 老鼠和人类。这些基因在很大程度上是未知的；而且， 它们的基因产物和作用机制尚不清楚。 隐性 MRL 小鼠中发生的突变 lpr 导致了深远的影响 淋巴结病伴有不寻常的 T 细胞亚群扩张和 全身性自身免疫性疾病。 提出了“反向”遗传方法 鉴定 lpr 基因座的突变或缺陷基因产物，以及 MRL 基因与 lpr 相互作用导致肾脏疾病。 方法 依赖于定义这些基因的物理位置。 RFLV 和 微卫星多态性将被用作两个不同的标记 杂交分离对临床至关重要的隐性基因 以及这些小鼠的血清学疾病。 对肾脏组织学进行评分 并与基因型分析和自身抗体谱相关。 这 lpr 对小鼠 Chr 19 的定位将得到证实并饱和 使用来自流式排序小鼠 Chr 19 文库的克隆进行映射将是 执行。 最令人感兴趣的是定义 MRL 数量性状 易患肾炎的位点（QTL）。 每个的详细映射 将使用可用的克隆来进行基因组的相关区域 定位于特定的染色体片段和克隆可能位于 基因组的那个区域。 如有必要，可进行亚染色体特异性检测 将生成文库或染色体显微切割文库 允许对已识别的 QTL 区域之一进行饱和作图。 长的 小鼠相关区域的范围限制性位点分析 将进行基因组分析。 如果（很可能）存在一个或多个自身免疫位点 位于小鼠和小鼠之间很大程度上保守的染色体片段内 人类基因组，这些研究可能会确定染色体的位置 人类的“自身免疫基因”。 多种策略，包括使用 染色体跳跃和小鼠酵母人工染色体文库， 除了长范围限制映射之外，还将用于更精确地 定义负责的关键基因的分子位置 遗传倾向。 与 MRL 正负/正负小鼠的比较 对于确定 lpr 突变以及比较 CBA/KJ 在指定菌株的 lpr 基因座上有新突变 CBA/KJ-lpr(cg) .对于其他局部“自身免疫基因”，候选基因 可以通过它们的染色体位置来暗示。 单一的定义 易患自身免疫的基因应该能够提供基本的洞察力 深入研究小鼠和人类疾病的分子基础。