HLA-DQ GENE COMPLEMENTATION IN LUPUS

狼疮中的 HLA-DQ 基因互补

• 批准号: 3137911
• 负责人: Courtney Montgomery
• 金额: $ 12.91万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1993-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3137911
• 关键词: DNA; Sjogren's syndrome; alleles; autoantibody; autoimmune disorder; cell bank /registry; gene complementation; genetic polymorphism; genetic regulation; genome; haploidy; heterozygote; human subject; laboratory mouse; major histocompatibility complex; monoclonal antibody; oligonucleotides; plasma; restriction fragment length polymorphism; serum; systemic lupus erythematosus

Polymorphic genetic differences between individuals contribute to the specificity of immune responsiveness. The original observation leading to the present studies has been the association of the DQI/DQ2 heterozygote state with high levels of anti-Ro/SSA autoantibodies which has since been confirmed in three studies of systemic lupus erythematosus. The goals of this project since last review have been to determine whether a gene complementation model could be confirmed at the DNA level and to establish a clinical resource composed of clinical data, serum, plasma, DNA, and transformed cell lines to test this hypothesis as well as a general model of disease heterogeneity. Substantial progress has been demonstrated. A restriction fragment length polymorphism study of the HLA-DQ-alpha, DQ- beta, and DR-beta genes has demonstrated that the combined DQl and DQ2 association with anti-Ro/SSA could be attributed to DQ-alpha and DQ-beta genes, respectively. This result establishes gene complementation at HLA-DQ as a mechanism for anti-Ro/SSA production in these patients. An additional separable effect at DQ-beta has also been described. Extension of this study to the T cell receptor beta gene has described restriction fragments which are closely associated with anti-Ro/SSA in the context of the identified histocompatibility alleles. In addition, this work has led to a theory of disease heterogeneity in systemic lupus erythematosus. In this model, once an individual is potentiated for the disease, the autoantibodies produced are strongly influenced by genetic factors including the DQ1/DQ2 heterozygous state for anti-Ro/SSA. Subsequently, the combination of autoantibodies influences the expression of disease found in individual patients. The plan for future experiments is to first test our recent findings in a larger patient group using analogous methods. In addition, the HLA-DQ genes associated with the anti-Ro/SSA response will be identified by allele-specific oligonucleotides and in selected groups DQ genes will be sequenced. Finally, two approaches will be explored to test the hypothesis that antigen presentation of Ro/SSA occurs through the defined DQ-alpha and DQ-beta elements, that the T cell recognizes this autoantigen-histocompatibility complex, and that autoantibody production results. First, in vitro cell culture techniques and an HLA-DQ transfected mouse cell line will be used to establish the HLA-DQ structural requirements. Second, a mouse model in which human grafts produce anti-Ro/SSA may also be suitable. In this model, immunization with small amounts of purified Ro/SSA antigen leads to a substantial rise of the human anti-Ro/SSA autoantibodies produced by these mice. Overall, these experiments will define the contribution to disease heterogeneity from HLA-DQ and will identify and test the molecular and structural features of DQ-alpha and DQ-beta which are important in gene complementation for anti- Ro/SSA.

个体之间的多态遗传差异有助于 免疫反应性的特异性。最初的观察导致了 目前的研究是DQI/DQ2杂合子的关联 具有高水平抗Ro/SSA自身抗体的状态，自那以来 在系统性红斑狼疮的三项研究中得到证实。的目标 这个项目自上次审查以来一直是为了确定一个基因 互补模型可以在DNA水平上得到证实，并建立 由临床数据、血清、血浆、DNA和 转化的细胞系来检验这一假设以及一个通用的模型 疾病的异质性。已经取得了实质性的进展。一个 人类白细胞抗原-DQ-α、DQ-α的限制性片段长度多态性研究 和DR-β基因已经证明DQ1和DQ2结合在一起 抗-Ro/SSA的相关性可能归因于DQ-α和DQ-β 分别是基因。这一结果确立了人类白细胞抗原-DQ的基因互补 作为这些患者产生抗Ro/SSA的一种机制。一项额外的 还描述了DQ-Beta的分离效应。这是它的延伸 对T细胞受体β基因的研究描述了限制性片段 它们与抗Ro/SSA密切相关 确定了组织相容等位基因。此外，这项工作还导致了 系统性红斑狼疮的疾病异质性理论。在这 模型，一旦一个人被强化了这种疾病， 自身抗体的产生受到遗传因素的强烈影响。 包括抗Ro/SSA的DQ1/DQ2杂合态。随后， 自身抗体的组合影响在 个别病人。未来实验的计划是首先测试我们的 使用类似方法在一个更大的患者组中的最新发现。在……里面 此外，与抗Ro/SSA反应相关的HLA-DQ基因将是 由等位基因特异的寡核苷酸鉴定，并在选定的DQ组中 基因将被测序。最后，将探索两种方法来进行测试 假设Ro/SSA的抗原提呈是通过 定义的DQ-α和DQ-β元素，T细胞识别这一点 自身抗原-组织相容性复合体以及自身抗体产生 结果。首先，体外细胞培养技术和人类白细胞抗原-DQ的转染法 将小鼠细胞系用于建立人类白细胞抗原-DQ结构 要求。第二，人类移植产生的小鼠模型 抗Ro/SSA抗体也可能适用。在这个模型中，用小剂量免疫 纯化的Ro/SSA抗原的量导致人类 这些小鼠产生的抗Ro/SSA自身抗体。总的来说，这些 实验将定义以下因素对疾病异质性的贡献 并将鉴定和测试分子和结构特征 DQ-α和DQ-β在抗病毒基因互补中的重要作用 RO/SSA。