HLA-DP SEQUENCE POLYMORPHISM AND AUTOIMMUNITY

HLA-DP 序列多态性与自身免疫

• 批准号: 3143751
• 负责人: HENRY A ERLICH
• 金额: $ 9.05万
• 依托单位: HOFFMAN-LA ROCHE, INC.--ROCHE MOLECULAR SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-03 至 1993-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3143751
• 关键词: African; Arabs; MHC class II antigen; South American; autoimmune disorder; celiac disease; genetic manipulation; genetic polymorphism; genetic recombination; graft versus host disease; human genetic material tag; human population genetics; human tissue; insulin dependent diabetes mellitus; juvenile rheumatoid arthritis; linkage mapping; major histocompatibility complex; multiple sclerosis; myasthenia gravis; nucleic acid probes; pemphigus; polymerase chain reaction; rheumatoid arthritis; western blottings

The overall goal of this project is to increase our understanding of the HLA-linked susceptibility to autoimmune diseases. A variety of autoimmune disorders have been associated with serologically defined variants at the HLA class ll loci. Recently, molecular analyses of class II polymorphisms have revealed the heterogeneity within these serotypes and suggested that structural variation in the class ll products of HLA-DR and -DQ may confer disease susceptibility. The structures of the DPalpha and DPbeta genes as well as their role in disease susceptibility, however, have not yet been thoroughly investigated. We propose to use the polymerase chain reaction (PCR) method to amplify the DPalpha and DPbeta genes from a variety of individuals. Characterization of the alleles will initially be done by sequence analysis. Based on these sequences, non-isotopic oligonucleotide probes will be synthesized and used in a dot blot format to rapidly screen a large number of individuals from different ethnic groups to ascertain allele frequencies and identify new alleles. The role of DP in disease susceptibility will be investigated using the same dot blot format on patient populations as well as the appropriate controls. The autoimmune diseases investigated will include juvenile rheumatoid arthritis, adult rheumatoid arthritis, multiple sclerosis, celiac disease, myasthenia gravis, IDDM and Pemphigus vulgaris. All samples will also be DNA-typed for HLA-DR and -DQ since, based on our previous work, disease susceptibility appears most highly associated with specific combinations of class ll alleles. In order to determine whether a particular disease-associated DP allele actually contributes to susceptibility independent of other HLA-D region alleles or whether it simply "marks" an extended "disease" haplotype we will need to determine whether any of the DP alleles are in linkage disequilibrium with a particular HLA haplotype. We will therefore examine the association of DR, DQ, and DP types in both family studies and in homozygous typing cells. The recombination rate between DP and DR/DQ will also be measured by utilizing the ability of PCR to co-amplify two genes, here DPbeta and DQalpha, from single sperm. The role of DP polymorphism in graft vs. host disease will be determined by DP-typing 200 bone marrow recipients and their apparently "HLA"-matched donor sibs, to determine whether differences at the DP locus correlate with occurrence of graft vs. host disease. Finally, we will look at the evolution of polymorphism at the DP locus by characterizing its allelic polymorphism in other primates. These studies will not only contribute to our understanding of the structure of the DP molecule and its role in disease susceptibility but also to its importance in the biology of tissue transplantation.

这个项目的总体目标是增加我们对 人类白细胞抗原与自身免疫性疾病的易感性有关。各种自身免疫性疾病 疾病与血清学定义的变异体有关 人类白细胞抗原分类和基因定位。最近，II类基因多态的分子分析 揭示了这些血清型的异质性，并表明 人类白细胞抗原-DR和-DQ类产物的结构变异可能会导致 疾病易感性。DPalpha和DPbeta基因的结构 以及它们在疾病易感性中的作用，然而，还没有 彻底调查过了。我们建议使用聚合酶链式反应 扩增DPalpha和DPbeta基因的聚合酶链式反应(PCR)方法 个人。等位基因的表征最初将通过以下方式完成 序列分析。基于这些序列，非同位素寡核苷酸 将合成探针并以斑点杂交的形式使用，以快速筛选 来自不同种族的大量个人来确定 等位基因频率和识别新的等位基因。DP在疾病中的作用 将使用相同的斑点印迹格式研究敏感度 患者群体以及适当的对照。自身免疫 调查的疾病将包括幼年类风湿性关节炎，成人 类风湿性关节炎、多发性硬化症、乳糜泻、肌无力 Gravis、IDDM和寻常型天疱疮。所有样本也将进行DNA分型 对于人类白细胞抗原-DR和-DQ，因为根据我们以前的工作，疾病 易感性似乎与特定的组合高度相关 11类等位基因。以确定是否有特定的 疾病相关的DP等位基因实际上增加了易感性 独立于其他人类白细胞抗原-D区等位基因，或者它是否只是“标记”一种 扩展的“疾病”单倍型我们将需要确定是否有任何 DP等位基因与特定的人类白细胞抗原单倍型存在连锁不平衡。 因此，我们将检查DR、DQ和DP类型在这两种类型中的关联 家系研究和纯合子分型细胞。重组率 还将利用聚合酶链式反应的能力来测量DP和DR/DQ之间的关系 从单个精子中共同扩增两个基因，这里是DPbeta和DQpha。这个 DP多态在移植物抗宿主疾病中的作用将通过以下方式确定 200例骨髓移植受者及其明显“人类白细胞抗原”相合的DP分型 供体同胞，以确定DP基因座的差异是否与 移植物对宿主疾病的发生。最后，我们将看看 应用等位基因研究DP基因座的多态进化 其他灵长类动物的多态现象。这些研究不仅将有助于 我们对DP分子结构的理解及其在 疾病的易感性，以及它在组织生物学中的重要性 移植。