Genetic Mechanisms to Suppress Autoimmunity

我们建议确定Slesl，一种上位性修饰物，在我们的B6-B6中抑制致命狼疮的发展- 系统性自身免疫的同源模型。在初步研究中，我们已经将该基因定位到956kb 小鼠17号染色体上的同源间隔，并开发了一种允许表型检测的策略 在大约3个月龄的小鼠身上发现了Slesl。我们现在建议鉴定这个基因并描述其分子特征。 它调节抑制致命性狼疮的通路和细胞谱系。我们有两个具体目标： 具体目标1.定位和识别Slesl。我们将Slesl定位到&lt；200 KB的基因组片段 通过使用表达一系列86来源BAC(细菌人工)的转基因小鼠进行表型拯救 染色体)跨越临界区。Sles1对自身免疫的抑制在杂交中是隐性的 对于B6，表明含有Slesl基因座的66个衍生袋子会导致B6.Sle1Sleslyaa小鼠 发展自身免疫力。总共需要7个B6-BAC转基因菌株才能跨越Slesl关键 间隔，其中一种将含有SLEL，并导致B6.Sle1SLEL小鼠的自身免疫表型。这个 将详细分析该BAG中候选基因的基因组特征，并将Slesl 通过体内分析确定，使用BAC修饰技术扰乱已验证的候选对象 基因位于袋子里。 明确SLEL的分子途径和免疫学机制 抑制疾病。我们正在进行的对Slesl的分析表明，该基因调节表型 表达于B淋巴细胞、T淋巴细胞和单核细胞。每一支血统在压抑 Slesl的自身免疫将通过过继转移和/或混合骨髓嵌合体来确定。在……里面 此外，我们将利用Illumina Mouse-6珠芯片进行全球基因表达分析，以确定基因 在这些谱系中，被Slesl修改的通路。这些分析将描述 介导SLEL抑制自身免疫的免疫学机制并提供重要的新的 对调节免疫耐受和抑制启动剂的免疫过程的见解 自身免疫力。

We propose to identify Slesl, an epistatic modifier that suppresses the development of fatal lupus in our B6- congenic model of systemic autoimmunity. In preliminary studies, we have localized this gene into a 956 Kb congenic interval on murine chromosome 17 and developed a strategy that allows the phenotypic detection of Slesl in mice at ~3 months of age. We now propose to identify this gene and characterize the molecular pathways and cell lineages that it modulates to suppress fatal lupus. We have two specific aims: Specific aim 1. To localize and identify Slesl. We will localize Slesl to a genomic segment of < 200 Kb via phenotypic rescue using transgenic mice expressing a series of 86-derived BACs (bacterial artificial chromosomes) spanning the critical region. Suppression of autoimmunity by Slesl is recessive in crosses with B6, indicating that a 66-derived BAG containing the Slesl locus will cause B6.Sle1Sleslyaa mice to develop autoimmunity. A total of 7 B6-BAC transgenic strains will be required to span the Slesl critical interval, one of which will contain Slesl and cause autoimmune phenotypes in B6.Sle1Slesl mice. The genomic characteristics of the candidate genes within this BAGwill be analyzed in detail and Slesl will be definitively identified via in vivo analysis using BAC-modification technology to disrupt validated candidate genes located in the BAG. Specific aim 2. To define the molecular pathways and immunologic mechanisms by which Slesl suppresses disease. Our ongoing analysis of Slesl indicates that this gene modulates phenotypes expressed in B lymphocytes, T lymphocytes, and monocytes. The role of each lineage in the suppression of autoimmunity by Slesl will be determined by adoptive transfer and/or mixed bone marrow chimeras. In addition, we will utilize the Illumina Mouse-6 BeadChip for global gene expression analysis to identify genetic pathways that are modified by Slesl in each of these lineages. These analyses will characterize the immunologic mechanisms that mediate the suppression of autoimmunity by Slesl and provide important new insights into the immunologic processes that regulate immune tolerance and suppress incipient autoimmunity.