Genetic Mechanisms to Suppress Autoimmunity

我们建议确定Slesl，一种上位性修饰剂，可以抑制我们的B6细胞中致命性狼疮的发展。 系统性自身免疫的同类模型。在初步研究中，我们已经将该基因定位于956 Kb的 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 在约3月龄的小鼠中的Slesl。我们现在建议鉴定该基因并表征其分子特征。 它调节的途径和细胞谱系来抑制致命的狼疮。我们有两个具体目标： 具体目标1.定位和识别Slesl。我们将Slesl定位于< 200 Kb的基因组片段， 通过使用表达一系列86-衍生的BAC（细菌人工 染色体）跨越临界区域。Slesl对自身免疫的抑制在杂交中是隐性的 与B6，表明含有Slesl基因座的66衍生BAG将导致B6.Sle1Sleslyaa小鼠 产生自身免疫将需要总共7个B6-BAC转基因菌株来跨越Slesl关键菌株。 间隔，其中之一将含有Slesl并在B6.SlelSlesl小鼠中引起自身免疫表型。的 将详细分析该BAG内候选基因的基因组特征，并将Slesl 通过使用BAC修饰技术的体内分析明确鉴定，以破坏经验证的候选物 位于BAG中的基因。 具体目标2。明确Slesl感染的分子途径和免疫机制， 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 在B淋巴细胞、T淋巴细胞和单核细胞中表达。每个谱系在抑制 通过过继转移和/或混合骨髓嵌合体来确定Slesl的自身免疫。在 此外，我们将利用Illumina Mouse-6 BeadChip进行全球基因表达分析，以识别遗传物质 在这些谱系中的每一个中被Slesl修饰的途径。这些分析将描述 免疫机制，介导抑制自身免疫的Slesl，并提供重要的新的 对调节免疫耐受和抑制早期免疫反应的免疫学过程的见解 自身免疫

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.