HLA-DQ GENE COMPLEMENTATION IN LUPUS

狼疮中的 HLA-DQ 基因互补

• 批准号: 3137906
• 负责人: Courtney Montgomery
• 金额: $ 12.26万
• 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 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3137906
• 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.

个体之间的多态性遗传差异有助于 免疫反应的特异性。最初的观察结果导致 目前的研究表明，DQI/DQ 2杂合子 状态与高水平的抗Ro/SSA自身抗体， 在三项系统性红斑狼疮的研究中得到证实。的目标 这个项目自上次审查以来一直在确定一个基因是否 互补模型可以在DNA水平上得到证实， 由临床数据、血清、血浆、DNA组成的临床资源，以及 转化的细胞系来测试这一假设以及一个通用模型 疾病的异质性。已经取得了实质性进展。一 限制性片段长度多态性研究HLA-DQ-α，DQ- 已经证明，组合的DQ 1和DQ 2基因可以在细胞内表达。 与抗Ro/SSA的相关性可归因于DQ-α和DQ-β 基因，分别。这一结果确立了HLA-DQ的基因互补 作为这些患者产生抗Ro/SSA的机制。额外 还描述了DQ-β的可分离效应。延长这一 对T细胞受体β基因的研究描述了限制性片段 与抗Ro/SSA密切相关， 鉴定了组织相容性等位基因此外，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，一旦一个人对疾病增强， 产生的自身抗体受遗传因素的强烈影响 包括抗Ro/SSA的DQ 1/DQ 2杂合状态。随后 自身抗体的组合会影响疾病的表达， 个别患者。未来的实验计划是首先测试我们的 最近的研究结果在一个更大的病人群体使用类似的方法。在 此外，与抗Ro/SSA反应相关的HLA-DQ基因将被 通过等位基因特异性寡核苷酸鉴定， 基因将被测序。最后，将探索两种方法来测试 假设Ro/SSA的抗原呈递是通过 定义的DQ-α和DQ-β元件，T细胞可以识别这些元件， 自身抗原-组织相容性复合物，以及自身抗体产生 结果首先，在体外细胞培养技术和HLA-DQ转染 小鼠细胞系将用于建立HLA-DQ结构 要求.第二，一个小鼠模型， 抗Ro/SSA也可以是合适的。在这个模型中，免疫接种小 大量纯化的Ro/SSA抗原导致人免疫球蛋白的表达显著增加。 这些小鼠产生的抗Ro/SSA自身抗体。总的来说，这些 实验将定义疾病异质性的贡献， HLA-DQ，并将确定和测试的分子和结构特征， DQ-α和DQ-β在抗-HCV基因互补中是重要的。 Ro/SSA。