HLA-DP SEQUENCE POLYMORPHISM AND DISEASE PREDISPOSITION

HLA-DP 序列多态性与疾病易感性

• 批准号: 2064803
• 负责人: HENRY A ERLICH
• 金额: $ 12.92万
• 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 1997-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2064803
• 关键词: AIDS; African; Arabs; Asians; European; Hispanic Americans; Hodgkin's disease; MHC class II antigen; Native Americans; Scandinavian; South American; autoimmune disorder; cervix neoplasms; disease /disorder proneness /risk; family genetics; genetic polymorphism; human genetic material tag; human tissue; inflammatory bowel diseases; insulin dependent diabetes mellitus; linkage mapping; major histocompatibility complex; multiple sclerosis; nucleic acid probes; polymerase chain reaction; racial /ethnic difference; sperm; western blottings

The overall goal of this project is to determine the nature and extent of allelic diversity in the HLA-DP region and to assess the relationship of this polymorphism to HLA-associated disease susceptibility. A variety of diseases, many but not all of which are autoimmune, have been associated with specific serologically defined HLA class II alleles and/or haplotypes. Recently, DNA-based typing has shown that particular DR and DQ sequence-defined alleles were associated with specific diseases; sequence comparisons of susceptible and non-susceptible alleles revealed the potential importance of individual polymorphic positions of the class II beta chains. The role of DP polymorphism in disease susceptibility has not been studied to the same extent. The distribution of DPB1 and DPA1 alleles in patients and in ethnically matched controls will be compared for a variety of diseases. Due to the strong linkage disequilibrium within the HLA region, assessing the role of DP polymorphism requires the analysis of HLA (B,DR,DQ,DP) haplotypes . We have already examined the nature and distribution of extended haplotypes in a Caucasian group. Our project will focus on families, allowing the unambiguous determination of haplotypes and we will compare the different haplotypic combinations that are found in different ethnic groups. These haplotype data are critical not only for understanding the complex patterns of HLA-disease associations but also for defining the likelihood of finding HLA-matched bone marrow and solid organ donors in different ethnic groups. Our proposal to characterize the DP sequence polymorphism, to refine our non-radioactive oligonucleotide probe DP typing system, and to apply this method (both dot blot and reverse dot blot) to disease susceptibility studies is based upon the polymerase chain reaction (PCR). Our previous work using DPB1 and DPA1 primers to PCR amplify the polymorphic second exon for sequence analysis will be extended to additional samples and populations to identify new alleles, revealed as a novel pattern of probe reactivity. New alleles are always confirmed by sequence analysis. The relationship of recombination frequency to the observed patterns of linkage disequilibrium will be examined in family studies but also by using the approach of sperm mapping, the co-amplification by PCR of linked loci from individual sperm. This method for determining recombination frequencies over short genetic distances, like those within the HLA region, can be used to compare recombination frequencies between individuals and between haplotypes. PCR amplification from sperm will also be used to try to detect the generation of new DPB1 alleles by segmental exchange, the putative gene-conversion-like mechanism inferred from the patchwork pattern of DPB1 polymorphism. A phylogenetic analysis of DPB1 polymorphism will also be carried out on a data set of non-human primate DPB1 alleles. These studies will increase our knowledge of HLA-DP polymorphism and contribute to our understanding of its role in disease susceptibility.

该项目的总体目标是确定 HLA-DP 区域的等位基因多样性并评估 这种多态性与 HLA 相关疾病的易感性有关。 各种 许多（但并非全部）疾病与自身免疫性疾病有关 具有特定血清学定义的 HLA II 类等位基因和/或 单倍型。 最近，基于 DNA 的分型表明，特定的 DR 和 DQ 序列定义的等位基因与特定疾病相关； 易感和非易感等位基因的序列比较揭示 该类个体多态性位置的潜在重要性 II β链。 DP多态性在疾病易感性中的作用 尚未得到同等程度的研究。 DPB1和的分布 患者和种族匹配对照中的 DPA1 等位基因将 对多种疾病进行比较。 由于关联性强 HLA 区域内的不平衡，评估 DP 的作用 多态性需要分析 HLA（B、DR、DQ、DP）单倍型。我们 已经检查了扩展单倍型的性质和分布 在一个白人群体中。 我们的项目将重点关注家庭，让 单倍型的明确确定，我们将比较不同的 在不同种族群体中发现的单倍型组合。 这些 单倍型数据不仅对于理解复杂的 HLA-疾病关联的模式，也可用于定义可能性 寻找 HLA 匹配的骨髓和实体器官捐献者 族裔群体。 我们关于表征 DP 序列的建议 多态性，以完善我们的非放射性寡核苷酸探针 DP 打字系统，并应用此方法（点印迹和反向点印迹） 印迹）疾病敏感性研究是基于聚合酶 链式反应（PCR）。 我们之前的工作使用 DPB1 和 DPA1 引物 PCR 扩增多态性第二外显子进行序列分析 扩展到更多样本和群体以识别新的等位基因， 揭示了一种新的探针反应模式。 新的等位基因总是存在 通过序列分析证实。 重组关系 观察到的连锁不平衡模式的频率将是 在家庭研究中进行了检查，但也通过使用精子的方法进行了检查 作图，通过 PCR 联合扩增个体的连锁基因座 精子。 这种方法可以确定短期内的重组频率 遗传距离，如 HLA 区域内的距离，可用于 比较个体之间和个体之间的重组频率 单倍型。 精子的 PCR 扩增也将用于尝试 通过分段交换检测新 DPB1 等位基因的生成， 从拼凑中推断出的假定的类似基因转换的机制 DPB1 多态性模式。 DPB1的系统发育分析 还将对非人类灵长类动物的数据集进行多态性分析 DPB1 等位基因。 这些研究将增加我们对 HLA-DP 的了解 多态性并有助于我们了解其在疾病中的作用 易感性。