DEVELOPMENT OF MUCOSAL IMMUNITY TO COMMENSAL ORAL BACTERIA

对口腔共生细菌的粘膜免疫的发展

• 批准号: 3803741
• 负责人: MICHAEL COLE
• 金额: --
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803741
• 关键词: Actinomyces; Bacteroides; Streptococcus agalactiae; Streptococcus mutans; antibody specificity; autoradiography; bactericidal immunity; breast feeding; enzyme linked immunosorbent assay; gel electrophoresis; growth /development; high performance liquid chromatography; human milk; human subject; immunity; immunopathology; infant human (0-1 year); laboratory mouse; longitudinal human study; microorganism culture; microorganism growth; monoclonal antibody; oral bacteria; oral mucosa; passive immunization; radiotracer; saliva; secretory immune system; surface antigens

The goal of this proposal is to determine the role of the developing salivary immune response to colonization of mucosal surfaces by selected commensal oral bacteria. The normal oral flora comprises many bacterial species, some are pathogenic producing diseases ranging from chronic low-grade infection of tooth and periodontium, to acute, systemic, life-threatening sepsis, meningitis and pneumonia. Although the salivary immune system recognizes and responds to oral bacteria the extent to which it plays a role in the regulation of the indigenous oral flora is unclear. An understanding of the mechanisms by which host protection is mediated against commensal bacteria with pathogenic potential is essential to the development of vaccines for their control. The objective of this proposal is to determine the relationship between the levels, isotypes and specificities of salivary antibodies and the colonization of selected oral bacteria. The bacteria chosen for study are Streptococcus mutans, S. agalactiae (GBS), Actinomyces viscosus, A. naeslundii and Bacteroides intermedius. These bacteria were selected because they have been implicated in chronic low-grade infection (dental caries and periodontitis) or severe acute sepsis and they represent genera that are either unique to the mouth or are oral species of genera that predominate at other mucosal surfaces. It is proposed to conduct a longitudinal study in infants from birth to age two years. The specific aims of this study are to (1) determine the kinetics of colonization of S. mutans, A. viscosus, A. naeslundii, B. intermedius and GBS using selective media and fluorescent antibodies, (2) determine the levels and isotypes of salivary antibodies induced in response to colonization of the selected bacteria using an enzyme-linked immunosorbent assay (ELISA) (3) determine the specificity of antibodies induced in response to colonization by Western blotting (4) identify and characterize immunodominant surface antigens using monoclonal antibodies and high performance liquid chromatography and (5) correlate the levels, isotypes and specificity of salivary antibodies with colonization of the selected bacteria by comparing ELISA titers and Western blot profiles with the appearance and number of the selected bacteria in the mouth. As immunity in the neonate's oral cavity may be influenced by maternal transfer of antibody in breast milk and through the placenta, the effect of passive immunity on colonization of the selected bacteria will also be addressed in Specific Aims 2-5.

该提案的目标是确定 对粘膜定植产生唾液免疫反应 由选定的共生口腔细菌表面。 正常的口腔 菌群包含许多细菌种类，其中一些是致病性的 产生各种疾病，从慢性低度感染 牙齿和牙周，急性、全身性、危及生命 败血症、脑膜炎和肺炎。 虽然唾液免疫 系统识别口腔细菌并对其做出反应的程度 它在调节本地口腔菌群中发挥作用 不清楚。 了解宿主的机制 保护是针对具有致病性的共生细菌介导的 潜力对于开发疫苗至关重要 控制。 该提案的目的是确定 水平、同种型和特异性之间的关系 唾液抗体和选定口腔细菌的定植。 选择用于研究的细菌是变形链球菌、链球菌。 无乳杆菌 (GBS)、粘性放线菌、内氏放线菌和 中间拟杆菌。 选择这些细菌是因为它们 与慢性低度感染（龋齿 和牙周炎）或严重急性败血症，它们代表属 要么是口腔独有的，要么是口腔属的物种 在其他粘膜表面占主导地位。 建议 对从出生到两岁的婴儿进行纵向研究 年。 本研究的具体目的是（1）确定 变形链球菌、粘性链球菌、内氏链球菌、内氏链球菌的定植动力学 B. 使用选择性培养基和荧光的中间体和 GBS 抗体，（2）测定唾液的水平和同种型 响应选定的定植而诱导的抗体 使用酶联免疫吸附测定 (ELISA) 检测细菌 (3) 确定响应所诱导的抗体的特异性 通过蛋白质印迹 (4) 鉴定和表征定植 使用单克隆抗体的免疫显性表面抗原和 高效液相色谱法和 (5) 关联 唾液抗体的水平、同种型和特异性 通过比较 ELISA 滴度和 蛋白质印迹分析，包括外观和数量 口腔中的特定细菌。 作为新生儿口腔的免疫力 空腔可能受到母体乳房中抗体转移的影响 牛奶和通过胎盘，被动免疫的影响 所选细菌的定植也将在 具体目标 2-5。