Genetic Mechanisms to Suppress Autoimmunity

抑制自身免疫的遗传机制

基本信息

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
项目类别：
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

项目摘要

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.

我们建议鉴定 Slesl，一种上位修饰剂，可抑制 B6- 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，我们已将该基因定位到 956 Kb 小鼠 17 号染色体上的同类间隔，并开发了一种允许表型检测的策略大约 3 个月大的小鼠中的 Slesl。我们现在建议鉴定该基因并表征其分子特征它调节抑制致命性狼疮的途径和细胞谱系。我们有两个具体目标：具体目标 1. 本地化并识别 Slesl。我们将 Slesl 定位到 < 200 Kb 的基因组片段通过使用表达一系列 86 衍生 BAC（细菌人工染色体）跨越关键区域。 Slesl 对自身免疫的抑制在杂交中是隐性的与 B6，表明包含 Slesl 位点的 66 衍生 BAG 将导致 B6.Sle1Sleslyaa 小鼠发展自身免疫。总共需要 7 个 B6-BAC 转基因菌株来跨越 Slesl 临界值间隔，其中之一将含有 Slesl 并在 B6.Sle1Slesl 小鼠中引起自身免疫表型。这该 BAG 内候选基因的基因组特征将被详细分析，Slesl 将被使用 BAC 修饰技术破坏经过验证的候选物，通过体内分析最终确定基因位于 BAG 中。具体目标 2. 明确 Slesl 的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。每个谱系在抑制中的作用 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。在此外，我们将利用 Illumina Mouse-6 BeadChip 进行全局基因表达分析，以鉴定遗传这些谱系中的每一个都被 Slesl 修饰的途径。这些分析将表征介导 Slesl 抑制自身免疫的免疫机制，并提供重要的新发现深入了解调节免疫耐受和抑制初期免疫反应的免疫过程自身免疫。