Cr2 as a Murine Lupus Susceptibility Gene

首席调查员/项目主任(最后、第一、中间)：Boackle，Susan A.描述：说明应用程序的广泛、长期目标和具体目标，并参考项目与健康的相关性。简明扼要地描述实现这些目标的研究设计和方法。避免总结过去的成就和使用第一人称。此摘要的目的是在脱离应用程序时，作为对拟议工作的简洁和准确的描述。如果申请得到资助，这一描述将成为公开信息。因此，不要将先知/机密信息混为一谈。不要超过所提供的空间。小鼠系统性红斑狼疮(SLE)的主要易感基因SLEL对应于3个独立影响NZM2410小鼠染色质耐受性的基因座。SLELC的同源区间由NZW衍生而来，含有编码补体受体I和2(CR1/CR2，CD35/CD21)的Cr2。CR1/CR2缺乏症在人类和动物模型中都与自身免疫性疾病有关。最近在SLELC CR1/CR2中发现了一个关键的配体结合域的结构差异，这导致了受体功能的显着损害。这些结果有力地支持了Cr2作为SLELC区间中的一个疾病易感基因的作用。本提案中概述的项目将针对NZW CR2在狼疮NZM2410小鼠模型中的作用进行表征。其具体目的是证明CR2是NZM2410 SLELC区间的狼疮易感基因，识别NZW CR2中与耐受性丧失有关的结构域，并确定NZW CR2导致耐受性丧失的机制。通过证明SLELC表型在正常基因产物的存在下分解，将提供CR2是SLELC基因座狼疮易感基因的证据。将对包含Cr2的重组菌株进行评估，以确保CR2功能障碍继续跟踪自身免疫性疾病，将使用导入重组蛋白的CR2缺陷细胞系以及表达各种形式多态NZW CR2的BAC转基因小鼠的B细胞来确定导致自身免疫表型的关键受体结构域。最后，改变的NZW CR2等位基因导致B细胞耐受性丧失的机制将用B细胞耐受性的3-83和HEL模型来表征。这些研究将阐明在NZM2410小鼠模型中受损的CR2的特定功能，这些功能可能会影响自身免疫性疾病的发展，从而成为治疗干预的重要靶点。表演网站========================================Section End===========================================

Principal Investigator/Program Director (Last, first, middle): Boackle, Susan A. DESCRIPTION: State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project. Describe concisely the research design and methods for achieving these goals. Avoid summaries of past accomplishments and the use of the first person. This abstract is meant to serve as a succinct and accurate description of the proposed work when separated from the application. If the application is funded, this description, as is, will become public information. Therefore, do not inctude prophetary/confidentiat information. DO NOT EXCEED THE ,SPACE PROVIDED. The major murine systemic lupus erythematosus (SLE) susceptibility locus, Slel, corresponds to 3 loci independently affecting loss of tolerance to chromatin in the NZM2410 mouse. The congenic interval corresponding to Slelc, derived from NZW, contains Cr2, which encodes complement receptors I and 2 (CR1/CR2, CD35/CD21). CR1/CR2 deficiency has been associated with autoimmune disease in both humans and in animal models. A structural difference in a critical ligand-binding domain has recently been identified in Slelc CR1/CR2 which results in significant impairment in receptor function. These results strongly support the role of Cr2 as a disease susceptibility gene in the Slelc interval. The project outlined in this proposal will be directed towards characterizing the role of NZW CR2 in the NZM2410 mouse model for lupus. The specific aims are to prove that CR2 is the lupus susceptibility gene in the NZM2410 Slelc interval, to identify the structural domains in NZW CR2 that are critical in loss of tolerance, and to determine the mechanisms by which NZW CR2 results in loss of tolerance. Proof that CR2 is the lupus susceptibility gene in the Slelc locus will be provided by demonstrating that the Slelc phenotypes resolve in the presence of normal gene products. Recombinant strains that contain narrowed intervals containing Cr2 will be assessed to ensure that CR2 dysfunction continues to track with autoimmune disease, The critical receptor domains that result in the autoimmune phenotypes will be determined, using both CR2-deficient cell lines transfected with recombinant proteins as well as B cells from BAC transgenic mice that express various forms of the polymorphic NZW CR2. Finally, the mechanisms by which the altered NZW CR2 allele results in loss of B cell tolerance will be characterized using the 3-83 and HEL models for B cell tolerance. These studies will clarify the specific functions of CR2, impaired in the NZM2410 mouse model, that may impact on the development of autoimmune disease and thus be important targets for therapeutic interventions. PERFORMANCE SITE ========================================Section End===========================================