SIGNIFICANT ANTIGENS OF SELECTED PERIODONTOPATHIC BACTERIA

选定的牙周病细菌的重要抗原

• 批准号: 6270296
• 负责人: Roy C Page
• 金额: $ 18.98万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6270296
• 关键词: Actinobacillus actinomycetemcomitans; Bacteroides; Bacteroides gingivalis; antibody specificity; antibody titering; antigen antibody reaction; antiidiotype antibody; bacterial antigens; bacterial vaccines; bactericidal immunity; enzyme linked immunosorbent assay; gel electrophoresis; human subject; human therapy evaluation; immunological substance; laboratory mouse; laboratory rabbit; lipopolysaccharides; monoclonal antibody; periodontitis; protein purification; serotyping

During the course of this Program Project, we have demonstrated that roughly half of patients with early-onset periodontitis are seronegative for antigens of putative periodontal pathogens, functionality of antibodies that are produced is unexpectedly low, and periodontal treatment results in seroconversion and production of biologically more effective antibodies. These observations, combined with our demonstration that immunization of M. fascicularis with a vaccine containing P. gingivalis significantly suppresses alveolar bone loss, has led us to propose to continue characterization of major cell envelope antigens of selected periodontopathic bacteria both biochemically and immunologically, and to continue development of a vaccine for human periodontal infections. In the upcoming grant period we will focus our efforts on the cell envelope antigens of A. actinomycetemcomitans, P. gingivalis, and B. forsythus, and on antigens shared by these species. We will complete characterization of the carbohydrate antigen of A. actinomycetemcomitans, and assess its potential for use in a vaccine. We will complete our ongoing efforts to purify and characterize the 200 kDa cell envelope protein antigen from B. forsythus. This preparation and the 43 kDa, 53 kDa, and 67 kDa envelope proteins and cysteine protease from P. gingivalis will be evaluated for LPS contamination using a highly sensitive fluorescence probe method, and contaminants removed chromatographically using HPLC and by isoelectric focusing. LPS from P. gingivalis and B. forsythus will be purified and separated into high and low molecular weight fractions. Purity and homogeneity of protein and carbohydrate antigens will be documented. These antigens will be used for measurement of antibody levels in patients before and following therapy, and provided to investigators in Project IV. Monoclonal antibodies will be generated in mice for LPS epitopes shared by P. gingivalis and B. forsythus for development and testing of anti-idiotype vaccines. A similar approach will be taken for the oligosaccharide antigen of A. actinomycetemcomitans. Protein antigens will be used to immunize rabbits. The resulting antibodies, as well as antibodies in sera of monkeys immunized with the whole-cell P. gingivalis vaccine, will be extensively studied with the aim of identifying the antigen(s) with the most potential for inducing protective immunity in the monkey model. Measurements will include titer, avidity, enhancement of chemiluminescence and phagocytosis and killing, and blocking activity. Successful completion of this project will provide the fundamental information and purified antigens necessary for development of a vaccine against periodontal infections.

在该计划项目的过程中，我们已经证明了 早发性牙周炎的患者中，大约有一半为血清症 对于推定牙周病原体的抗原， 产生的抗体意外低，牙周 治疗导致血清转化和生物的产生更多 有效的抗体。 这些观察结果与我们的 证明了用疫苗对筋膜分枝杆菌的免疫 含有牙龈疟原虫会显着抑制牙槽骨损失，已有 导致我们建议继续表征主要细胞包膜 选定的牙周病细菌的抗原均在生化和 免疫学，并继续开发人类的疫苗 牙周感染。 在即将到来的赠款期内，我们将集中精力 曲霉曲霉的细胞包膜抗原的努力，P。 牙龈和B. Forsythus，以及这些物种共享的抗原。 我们将完成A的碳水化合物抗原的表征。 放线菌，并评估其在疫苗中使用的潜力。 我们将完成净化和特征200的持续努力 B. forsythus的KDA细胞包膜蛋白抗原。 这项准备 以及43 kDa，53 kDa和67 kDa包膜蛋白和半胱氨酸 牙龈疟原虫的蛋白酶将用于使用LPS污染的蛋白酶 高度敏感的荧光探针方法，去除污染物 使用HPLC和等电聚焦在色谱上。 来自P的LPS 牙龈和B. Forsythus将被纯化并分为高和 低分子量分数。 蛋白质的纯度和同质性 将记录碳水化合物抗原。 这些抗原将被使用 用于测量患者前后患者的抗体水平 治疗，并提供给项目IV的研究人员。 单克隆 小鼠将生成抗体。 牙龈和。 疫苗。 寡糖将采取类似的方法 抗曲霉的抗原。 蛋白质抗原将用于 免疫兔子。 产生的抗体以及抗体 用全细胞牙龈疟原虫疫苗免疫的猴子血清将 进行广泛的研究，目的是确定抗原 在猴子模型中诱导保护性免疫的最大潜力。 测量将包括滴度，亲和力，增强 化学发光，吞噬作用和杀伤以及阻塞活性。 成功完成该项目将提供基本 开发疫苗所需的信息和纯化的抗原 反对牙周感染。