Role of SF Gene Cluster in Autoimmunity

超出所提供的空间。Sel同源间隔包含一簇启动自身免疫级联反应的基因，导致NZM 2410小鼠中的致命性狼疮。定位克隆分析已经鉴定了小鼠1号染色体上含有CD 150基因家族（CD 48、Lyl 08、Csl、CD 84、CD 244、CD 150、CD 229）的7个基因簇作为破坏对核抗原的耐受性的原因。这些基因调节许多淋巴细胞谱系中的激活阈值和效应子功能。我们比较了B6和B6.Slelb基因，发现该基因家族存在多个结构和调控多态性。我们提出这些多态性对免疫功能的影响如下：1）评估体内CD 150/CD 2家族基因的表达。将在疾病发作之前和之后从B6和B6.Slelb小鼠分离的各种淋巴细胞群中测定该基因簇的表达谱。这将通过流式细胞术、RT-PCR分析和在单个CD 150成员基因的调控元件的控制下产生具有报告构建体的转基因小鼠的组合来实现。2)描述CD 150家族基因多态性对淋巴细胞功能的影响。将对分离自B6和B6.SlelB小鼠的B和T细胞进行功能研究。这将使我们能够确定在CD 150基因簇的多态性对淋巴细胞功能的影响，在其他遗传相同的背景。3)建立一种评价CD 150家族成员功能的动物模型。该目的的目标是产生其中整个CD 150基因簇已经使用Cre重组酶和侧翼LoxP位点缺失的小鼠品系。这些小鼠对于研究该基因家族在体内淋巴细胞功能中的作用至关重要。此外，这些小鼠将用于通过BAC转基因拯救重建CD 150家族成员表达，以评估CD 150家族成员个体或组合在免疫功能中的作用。4)探讨CD 150基因簇多态性对自身免疫的影响。我们已经开发了B6衍生的（自身免疫抗性）和129 SV衍生的（自身免疫倾向）BAC重叠群，跨越CD 150基因簇，用于产生BAC转基因小鼠。这些将与CD 150簇缺失小鼠品系联合使用，以明确表征这些基因单独或联合的自身免疫潜力。性能现场=

EXCEED THE SPACE PROVIDED. The Slel congenic interval contains a cluster of genes that initiate the autoimmune cascade leading to fatal lupus in the NZM2410 mouse. Positional cloning analysis has identified a seven gene cluster on mouse chromosome 1 containing the CD150 family of genes (CD48, Lyl08, Csl, CD84, CD244, CD150, CD229) as causative for a breach in tolerance to nuclear antigens. These genes modulate activation thresholds and effector function in many lymphocyte lineages. Our comparison of B6 and B6.Slelb revealed the existence of several structural and regulatory polymorphisms in this gene family. We propose to characterize the impact of these polymorphisms on immune function as follows: 1) To assess the expression of the CD150/CD2 family of genes in vivo. Expression profiles of this gene cluster will be determined in various lymphoctye populations isolated from B6 and B6.Slelb mice prior to and following disease onset. This will be accomplished by a combination of flow cytometry, RT-PCR analysis and the generation of transgenic mice with reporter constructs under the control of regulatory elements of individual CD150 member genes. 2) To characterize the impact of CD150 family gene polymorphisms on lymphocyte function. Functional studies will be performed on B and T cells isolated from B6 and B6.Slelb mice. This will allow us to determine the impact of polymorphisms at the CD150 gene cluster on lymphocyte function in an otherwise genetically identical background. 3) To create an animal model to assess CD150 family member function. The goal of this aim is to generate a mouse strain in which the entire CD150 gene cluster has been deleted using Cre recombinase and flanking LoxP sites. These mice will be essential to study the role of this gene family in lymphocyte function in vivo. In addition, these mice will be used to reconstitute CD150 family member expression by BAC transgenic rescue to assess the role of individual or combinations of CD150 family members in immune function. 4) To assess the impact of the CD150 cluster polymorphisms on autoimmunity. We have developed B6-derived (autoimmune resistant) and 129Sv-derived (autoimmune prone) BAC contigs spanning the CD150 gene cluster for the generation of BAC transgenic mice. These will be used in combination with the CD150 cluster deletion mouse strain to definitively characterize the autoimmune potential of these genes singly or in combination. PERFORMANCE SITE ========================================Section End===========================================