Functional anaylses of the CDS48 and SLAMF8 receptors ........

CDS48 和 SLAMF8 受体的功能分析........................

• 批准号: 7135753
• 负责人: Arlene H. Sharpe
• 金额: $ 32.63万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7135753
• 关键词: B cell receptor; B lymphocyte; T cell receptor; T lymphocyte; antigens; bioengineering /biomedical engineering; cell surface receptors; disease /disorder onset; functional /structural genomics; gene deletion mutation; gene interaction; genetic screening; genetic susceptibility; genetically modified animals; genotype; helper T lymphocyte; immune tolerance /unresponsiveness; immunophenotype; laboratory mouse; leukocyte activation /transformation; natural killer cells; single nucleotide polymorphism; systemic lupus erythematosus; transfection

Genetic studies have implicated the SLAM family in Systemic lupus erythematosus (SLE), but the functional roles of specific gene(s) within the SLAM locus in SLE are not yet clear. Our recent studies show that CD48 deficient (-/-) mice backcrossed onto the C57BL/6 (B6) background from the 129Sv background for 12 generations develop a lupus-like syndrome. By 6 months of age, these CD48-/- mice develop activated T and B cells, anti-nuclear antibodies, immune complexes and glomerulonephritis. We also have found that there are defects in T cell tolerance in CD48-/- mice. There are 2 potential explanations for the development of the SLE phenotype: Disease could be caused by the absence of CD48, or due to epistatic interactions between 129Sv genes flanking the disrupted CD48 gene and B6 genes in the CD48-/- mouse strain. The goal of Project 3 is to investigate the role of CD48 in SLE. As a definitive tool to understand the essential functions of CD48, we will use CD48-/- B6 mice generated by homologous recombination using B6 ES cells. Our specific aims are: 1) To test the hypothesis that CD48 regulates antigen-specific CD4 T cell activation and tolerance. We will examine if there are intrinsic T cell defects and/or dysregulated interactions between APC and T cells. We also use CD244-/- mice to investigate whether the CD48 ligand, CD244, regulates T cell responses. This focus on CD244 is driven by genetic studies of Project 1 that implicate CD244 variation in human SLE and preliminary data that suggest CD244 on the APC may regulate T cell: APC interactions. 2) To test the hypothesis that CD48 regulates B cell activation and/or B cell tolerance. We will determine if there are B cell intrinsic defects, examine humoral immune responses, and generate CD48-/- VH3H9 transgenic mice to study the role of CD48 in regulating anti-dsDNA B cells. 3) To investigate the hypothesis that CD48 regulates the development of SLE. We determine whether SLE develops in B6 CD48-/- mice. If SLElike disease develops, then we will evaluate the function of CD48 on the T cell, B cell and other cell types in SLE. If SLE does not develop, we will test the hypothesis that CD48-/-(129Sv x B6)BC12 CD48- /- mice develop SLE due to flanking 129Sv SLAM family genes and not due to the disrupted CD48 gene, by introducing a 129Sv BAG with a normal or disrupted 129Sv CD48 gene into the CD48-/- C57BL/6 mouse and assess whether the resulting strains develop SLE. This PPG facilitates collaborative interactions and provides a number of important tools and approaches that will enable us to address these important issues. These studies should contribute to the overall goal of this PPG: to understand how SLAM family members control pathways that regulate the development of SLE.

遗传学研究表明SLAM家族与系统性红斑狼疮(SLE)有关，但 SLAM基因座中的特定基因(S)在系统性红斑狼疮中的作用尚不清楚。我们最近的研究 显示CD48缺陷(-/-)小鼠从129Sv回交到C57BL/6(B6)背景 背景：12代人发展成狼疮样综合征。到6个月大的时候，这些CD48-/-小鼠 发展活化的T和B细胞、抗核抗体、免疫复合体和肾小球肾炎。我们 还发现CD48-/-小鼠的T细胞耐受性存在缺陷。有两个潜力 对系统性红斑狼疮表型发展的解释：疾病可能是由于缺乏 CD48，或由于被破坏的CD48基因两侧的129Sv基因与B6之间的上位性相互作用 CD48-/-小鼠品系中的基因。项目3的目标是研究CD48在系统性红斑狼疮中的作用。作为一名 了解CD48基本功能的权威工具，我们将使用CD48-/-B6小鼠产生 利用B6 ES细胞进行同源重组。我们的具体目标是：1)检验假设 CD48调节抗原特异性的CD4T细胞的活化和耐受。我们会检查是否有 固有的T细胞缺陷和/或APC和T细胞之间的相互作用失调。我们也使用CD244-/- 研究CD48配体CD244是否调节T细胞反应。对CD244的关注 是由项目1的遗传学研究驱动的，该项目涉及人类SLE的CD244变异，并初步 数据表明APC上的CD244可能调节T细胞：APC的相互作用。2)检验假设 CD48调节B细胞活化和/或B细胞耐受。我们会确定是否有B细胞 先天缺陷，检查体液免疫反应，并产生CD48-/-VH3H9转基因小鼠 研究CD48对抗双链DNA B细胞的调节作用。3)研究CD48的假说 调节系统性红斑狼疮的发展。我们确定是否在B6 CD48-/-小鼠中发生SLE。如果睡意盎然 疾病的发展，我们将评估CD48对T细胞、B细胞和其他细胞的功能 输入SLE类型。如果SLE没有发展，我们将测试CD48-/-(129Sv X B6)BC12 CD48- /-小鼠发生系统性红斑狼疮是由于129Sv SLAM家族基因两侧，而不是由于CD48基因中断。 通过将含有正常或突变的129Sv CD48基因的129Sv包导入CD48-/-C57BL/6 并评估所产生的菌株是否会发展成系统性红斑狼疮。此PPG促进协作 并提供了许多重要的工具和方法，使我们能够解决 这些重要的问题。这些研究应该有助于本PPG的总体目标：理解 SLAM家族成员如何控制调节SLE发展的途径。