Genetic Mechanisms to Suppress Autoimmunity

抑制自身免疫的遗传机制

• 批准号: 7336587
• 负责人: Edward K. Wakeland
• 金额: $ 25.31万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7336587
• 关键词: Adoptive Transfer; Age-Months; Alleles; Autoimmune Process; Autoimmunity; B-Lymphocytes; Bacterial Artificial Chromosomes; Bacterial Chromosomes; Bone Marrow; Bone Marrow Transplantation; Candidate Disease Gene; Cell Lineage; Cell physiology; Cells; Characteristics; Chimera organism; Chromosomes, Human, Pair 17; Chromosomes, Human, Pair 2; Complement; Data; Defect; Detection; Development; Disease; Disruption; Family; Gene Expression; Genes; Genetic; Genomic Segment; Genomics; Goals; Haplotypes; Hematopoietic; Immune; Immune Tolerance; Immune system; Immunologics; Lesion; Localized; Lupus; Mediating; Modeling; Modification; Molecular; Monitor; Mus; Pathway interactions; Phenotype; Principal Investigator; Process; Protein Isoforms; Recombinants; Role; Series; T-Lymphocyte; TLR7 gene; Technology; Transgenic Mice; Transgenic Organisms; Y Chromosome; aged; anergy; cohort; congenic; histocompatibility gene; in vivo; insight; male; monocyte; programs; receptor; research study

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.

我们建议识别SLESL，这是一种抑制B6-中致命狼疮发展的上皮修饰符 系统自身免疫的先天模型。在初步研究中，我们将该基因定位于956 kb 鼠染色体17的持续间隔，并制定了一种允许表型检测的策略 大约3个月大的小鼠中的SLESL。现在，我们建议识别该基因并表征分子 途径和细胞谱系调节抑制致命的狼疮。我们有两个具体的目标： 特定目的1。要本地化并识别SLESL。我们将将SLESL定位到小于200 kb的基因组段 通过表型救援使用表达一系列86个衍生的BAC（细菌人工的转基因小鼠） 染色体）跨越关键区域。 SLESL抑制自身免疫性在十字架中是隐性的 使用B6，表明包含SLESL基因座的66个衍生的袋子会导致b6.sle1sleslyaa小鼠 发展自身免疫性。总共需要7 B6-BAC转基因菌株才能跨越SLESL临界 间隔，其中之一将包含SLESL并在B6.SLE1SLESL小鼠中引起自身免疫表型。这 详细分析该袋中候选基因的基因组特征，SLESL将是 通过使用BAC修饰技术通过体内分析确定确定验证的候选者 基因位于袋子中。 特定目的2。定义分子途径和免疫机制 抑制疾病。我们对SLESL的持续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。每个血统在抑制中的作用 SLESL的自身免疫性将由产物转移和/或混合骨髓嵌合体确定。在 此外，我们将利用Illumina Mouse-6 Beadchip进行全局基因表达分析来鉴定遗传 这些谱系中每个谱系中通过SLESL修改的途径。这些分析将表征 免疫机制介导SLESL抑制自身免疫的抑制并提供重要的新 对调节免疫耐受性并抑制初期的免疫过程的见解 自身免疫性。