Identifying Periodontal Antigens By Protein Microarray

通过蛋白质微阵列识别牙周抗原

• 批准号: 7095994
• 负责人: DOUGLAS S DARLING
• 金额: $ 17.94万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-19 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7095994
• 关键词: Actinobacillus actinomycetemcomitans; Bacteroides gingivalis; antigen antibody reaction; bacterial antigens; bacterial cytopathogenic effect; bacterial proteins; bacteriophage lambda; clinical research; dental disorder diagnosis; genetic library; genetic screening; host organism interaction; human tissue; humoral immunity; immune response; microarray technology; pathologic process; periodontium disorder; polymerase chain reaction; protein quantitation /detection; tissue /cell culture; virulence

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.

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。经过广泛的努力，已经鉴定出与牙周炎相关的相对较少的细菌抗原的基因。因此，牙周病期间宿主对口腔细菌的免疫反应尚未得到充分表征。分析口腔细菌和宿主细胞之间发生的分子相互作用是 NIDCR 的首要任务，并将有助于我们对牙周病的基本了解和临床工具。该项目的总体目标是定义在牙周病期间引发体液免疫反应的细菌蛋白（抗原），并确定不同抗体特异性与疾病严重程度或治疗反应的关系。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对治疗的反应有关。为了解决这一假设，我们建议使用细菌基因组 DMA 创建牙龈卟啉单胞菌和伴放线放线杆菌的噬菌体 lambda 表达文库。我们将用牙周炎患者和对照血清筛选文库以鉴定抗原肽。抗原蛋白将排列在微阵列载玻片上并用个体患者血清进行筛选。使用分层聚类分析，特定抗体或抗体组的存在将与患者的临床指数相关。这种方法利用了缺乏内含子的细菌基因组极高的基因密度以及构建微阵列的技术进步。 R21探索性资助是一个合适的机制，因为蛋白质组学尚未用于牙周疾病，并且需要开发细菌基因组表达文库和抗原微阵列。