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.

遗传研究已牵涉到全身性红斑狼疮（SLE）中的大满贯家族，但 SLE中猛击基因座中特定基因的功能作用尚不清楚。我们最近的研究 证明从129SV的C57BL/6（b6）背景上回到C57BL/6（B6）的CD48缺乏（ - - ）小鼠 12代的背景发展为狼疮综合征。到6个月大，这些CD48 - / - 小鼠 发展为活化的T和B细胞，抗核抗体，免疫复合物和肾小球肾炎。我们 还发现CD48 - / - 小鼠的T细胞耐受性缺陷。有2个潜力 SLE表型发展的解释：疾病可能是由于缺乏而引起的 CD48，或由于129SV基因之间的上皮相互作用，侧面破坏的CD48基因和B6 CD48 - / - 小鼠应变中的基因。项目3的目的是研究CD48在SLE中的作用。作为 确定CD48基本功能的确定工具，我们将使用CD48 - / - B6小鼠生成的CD48 - / - B6小鼠 使用B6 ES细胞的同源重组。我们的具体目的是：1）检验以下假设。 CD48调节抗原特异性CD4 T细胞活化和耐受性。我们将检查是否有 固有的T细胞缺陷和/或APC和T细胞之间的相互作用失调。我们还使用CD244 - / - 小鼠研究CD48配体CD244是否调节T细胞反应。关注CD244 由项目1的遗传研究驱动，该研究暗示了人类SLE和初步的CD244变化 暗示APC上CD244的数据可能会调节T细胞：APC相互作用。 2）检验假设 该CD48调节B细胞活化和/或B细胞耐受性。我们将确定是否有B单元 固有缺陷，检查体液免疫反应并生成CD48 - / - VH3H9转基因小鼠到 研究CD48在调节抗DSDNA B细胞中的作用。 3）研究CD48的假设 调节SLE的发展。我们确定SLE是否在B6 CD48 - / - 小鼠中发展。如果斜线 疾病发展，然后我们将评估CD48在T细胞，B细胞和其他细胞上的功能 SLE中的类型。如果SLE不发展，我们将测试CD48 - / - （129SV X B6）BC12 CD48-的假设 / - 小鼠由于129SV猛击的家庭基因而出现SLE，而不是由于CD48基因的破坏而引起的。 通过将带有正常或中断的129SV CD48基因的129SV袋引入CD48 - / - C57BL/6 鼠标并评估产生的应变是否发展为SLE。该PPG促进了协作 互动并提供许多重要的工具和方法，使我们能够解决 这些重要问题。这些研究应促进该PPG的整体目标：了解 家庭成员如何控制规范SLE发展的途径。