Identifying Periodontal Antigens By Protein Microarray

描述：牙周炎是对细菌病原体的慢性炎症反应，会对支撑牙齿的骨骼和软组织造成损害。广泛的努力已经确定了与牙周炎相关的相对较少的细菌抗原的基因。因此，宿主在牙周病过程中对口腔细菌的免疫反应还没有得到充分的表征。分析口腔细菌和宿主细胞之间的分子相互作用是NIDCR的高度优先事项，并将有助于我们对牙周病的基本理解和临床工具。该项目的总体目标是确定在牙周病期间引起体液免疫反应的细菌蛋白(抗原)，并确定不同的抗体特异性如何与疾病的严重程度或治疗反应相关。我们的假设是，针对某些细菌抗原组的抗体的存在与牙周病的严重程度或患者的治疗反应有关。为了解决这一假设，我们建议使用细菌基因组DMA来创建牙龈卟啉单胞菌和伴生放线杆菌的噬菌体lambda表达文库。我们将筛选牙周炎患者和对照血清的文库，以鉴定抗原肽。抗原蛋白将排列在微阵列载玻片上，并用个别患者血清进行筛选。使用分级聚类分析，特定抗体或抗体组的存在将与患者的临床指标相关。这种方法利用了细菌基因组极高的基因密度，没有内含子，以及构建微阵列的技术进步。R21探索性赠款是一个合适的机制，因为蛋白质组学还没有用于牙周病，需要开发细菌基因组表达文库和抗原微阵列。

DESCRIPTION: Periodontitis is a chronic inflammatory response to bacterial pathogens that causes damage to both bone and soft tissues that support the teeth. Extensive effort has identified the genes for relatively few bacterial antigens related to periodontitis. Therefore, the host immune response to oral bacteria during periodontal disease has not been adequately characterized. Analysis of the molecular interactions that occur between oral bacteria and host cells is a high priority of NIDCR, and will contribute to our basic understanding and clinical tools for periodontal disease. The overall goal of this project is to define the bacterial proteins (antigens) that elicit humoral immune responses during periodontal disease, and determine how the different antibody specificities relate to the severity of disease, or response to treatment. Our hypothesis is that the presence of antibodies to certain groups of bacterial antigens is associated with the severity of periodontal disease or the response to treatment in patients. To address this hypothesis, we propose to use bacterial genomic DMA to create bacteriophage lambda expression libraries for Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans. We will screen the libraries with periodontitis patient and control sera to identify antigenic peptides. The antigenic proteins will be arrayed on microarray slides and screened with individual patient serum. The presence of specific antibodies, or groups of antibodies, will be related to the clinical indices of the patient using hierarchical cluster analyses. This approach takes advantage of the extremely high gene density of the bacterial genome, which lacks introns, and of the technological advances for construction of microarrays. The R21 Exploratory Grant is an appropriate mechanism since proteomics have not been used for periodontal disease, and bacterial genomic expression libraries and antigen microarrays need to be developed.