HLA-DP SEQUENCE POLYMORPHISM AND DISEASE PREDISPOSITION

我们的目标是了解HLA-DP区域多态性的功能意义和演变及其与疾病易感性的关系。在其他HLA II类基因座，DRB 1和DQB 1的多态性已经牵连了一段时间在易感性的各种疾病。DPB 1基因座的广泛多态性（迄今已鉴定出>80个等位基因）在疾病易感性中的作用还不太清楚。我们的假设是，多个HLA基因座的等位基因的特定组合决定了对给定疾病的易感性程度。我们将集中在三种疾病：少关节幼年类风湿性关节炎，1型糖尿病和宫颈癌。这些疾病似乎与DPB 1相关，也与DRB 1和/或DQB 1基因座的特定等位基因相关。我们的高分辨率固定化探针分型方法的II类和I类基因座将被应用于患者和对照样品从各种人群。基于家族的材料允许分析单倍型共享、传递比和连锁不平衡模式以及分层分析，以查看DPB 1或任何其他个体HLA基因座处的一些相关等位基因是否赋予增加的风险或简单地反映与其他HLA基因座处的高风险等位基因的连锁不平衡。研究HLA等位基因多样性是如何进化的，以及它是如何在不同人群中分布的，可以提供对功能意义的见解。将通过使用基于PCR的方法测量精子中罕见变异DPB 1序列的频率来检验DPB 1和其他HLA基因座处多态性的拼凑模式反映基因转换（节段交换）的操作的假设，并将通过人类和非人类灵长类动物外显子2和相邻内含子序列的系统发育分析来分析DPB 1多样性的进化。将使用定量动力学PCR来监测细胞因子表达，以检查HLA疾病关联反映在特异性抗原刺激CD 4 +T细胞后促进Th 1和Th 2应答的差异趋势的假设。该方法将应用于HPV感染的宫颈样本和GAD肽刺激的体外系统，以研究HLA与宫颈癌和1型糖尿病的相关性。

Our goal is to understand the functional significance and evolution of the polymorphism at the HLA-DP region and its relationship to disease susceptibility. Polymorphism at other HLA class II loci, DRB1 and DQB1 has been implicated for some time in predisposition to a variety of diseases. The role of the extensive polymorphism at the DPB1 locus (>80 alleles identified thus far) in disease predisposition is much less well understood. It is our hypothesis that specific combinations of alleles at multiple HLA loci determine the extent of susceptibility to a given disease. We will focus on three diseases: pauciarticular juvenile rheumatoid arthritis, type 1 diabetes, and cervical carcinoma. These diseases appear to have DPB1 associations as well as associations with specific alleles at the DRB1 and/or DQB1 locus. Our high resolution immobilized probe typing methods for the class II and class I loci will be applied to patient and control samples from a variety of populations. Family based material allows the analysis of haplotype sharing, transmission ratios, and linkage disequilibrium patterns as well as stratification analysis to see whether some of the associated alleles at DPB1, or any other individual HLA locus, confer increased risk or simply reflect linkage disequilibrium with high risk alleles at other HLA loci. Studying how HLA allelic diversity has evolved and how it is distributed in various human populations can provide insights into functional significance. The hypothesis that the patchwork patterns of polymorphism at the DPB1 and at other HLA loci reflects the operation of gene conversion (segmental exchange) will be tested by using a PCR-based method to measure the frequency of rare variant DPB1 sequences in sperm and the evolution of DPB1 diversity will be analyzed via phylogenetic analysis of exon2 and adjacent intron sequences from human and non- human primates. The hypothesis that HLA disease associations reflect the differential tendency to promote Th1 and Th2 responses following specific antigen stimulation of CD4+T cells will be examined using quantitative kinetic PCR to monitor cytokine expression. This method will be applied to HPV-infected cervical samples and an in vitro system with GAD peptide stimulation to study the HLA associations with cervical cancer and type 1 diabetes.