INTERACTION OF ORAL STREPTOCOCCI WITH ANIMAL TISSUES

口腔链球菌与动物组织的相互作用

• 批准号: 3219588
• 负责人: MURRAY W STINSON
• 金额: $ 17.61万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-02-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219588
• 关键词: Streptococcus mutans; Streptococcus pyogenes; Streptococcus sanguis; antibody formation; bacterial antigens; bacterial endocarditis; binding proteins; cell wall; dental caries; dental caries vaccine; dental plaque; disease /disorder model; genetic strain; glomerulonephritis; heparan sulfate; heparin; histopathology; human subject; immune complex; immunization; immunoelectron microscopy; immunofluorescence technique; kidney cell; laboratory rabbit; microorganism culture; molecular pathology; monoclonal antibody; nucleic acid sequence; protein purification; proteoglycan; site directed mutagenesis; structural genes; teichoate; virulence

The goal of this research proposal is to identify and characterize the molecular mechanisms of oral streptococci that contribute to the pathogenesis of streptococcus-associated nephritides and to develop specific prophylactic and therapeutic measures for these diseases. We have shown that S. mutans, S. pyogenes and S. sanguis produce a heparin-- binding protein (HBP) that also binds to heparan proteoglycans in basal laminae of animal tissues, particularly in kidney, heart and liver. This cationic protein is loosely associated with the bacterial cell wall and is released along with other components, such as lipoteichoic acid (LTA), into the cell environment during growth. One specific aim is to define the pathogenic properties of purified HBP and LTA in rabbits particularly their abilities to mediate deposition in kidneys of circulating immune complexes and/or formation of immune complexes in situ. Kidney tissue will be analyzed for pathology using histologic and immunohistologic stains. A second aim is to evaluate antibody production to HBP and LTA by rabbits during immunization with nephritis-associated strains of streptococci or with other streptococcal strains. The relative quantities and properties of HBP on bacterial surfaces and in culture medium will be determined using monoclonal antibodies. Ultrastructural location of the antigens on streptococci will be determined using immunoelectron microscopy. The third aim is to isolate the structural genes of HBP from these bacteria and determine the nucleotide sequences to further characterize this virulence factor. The fourth aim is to generate HBP-deficient strains of streptococci, using insertion mutagenesis, and to compare them with isogeneic parental strains to prove the pathogenic properties of this protein. The information derived from the experiments proposed in this application will further our understanding of all infection-associated nephritides in humans and may ultimately permit the development of safe streptococcal vaccines to protect against infection without risk of tissue injury.

该研究建议的目的是识别和表征 口服链球菌的分子机制，有助于 链球菌相关的肾脏的发病机理，并发展 这些疾病的特定预防和治疗措施。 我们 已经表明，杂种链球菌，链球菌和sanguis产生肝素 - 结合蛋白（HBP）也与基础上的肝素蛋白聚糖结合 动物组织的薄片，特别是在肾脏，心脏和肝脏中。 这 阳离子蛋白与细菌细胞壁和 与其他成分一起释放，例如脂甲酸（LTA）， 在生长过程中进入细胞环境。 一个具体的目的是定义 兔子中纯化的HBP和LTA的致病特性尤其是 他们在循环免疫的肾脏中介导沉积的能力 原位免疫复合物的复合物和/或形成。 肾脏组织 将使用组织学和免疫组织学分析病理 污渍。 第二个目的是评估与HBP和LTA的抗体生产 兔子在免疫期间与肾炎相关菌株的免疫 链球菌或其他链球菌菌株。 亲戚 HBP在细菌表面和培养物上的数量和特性 培养基将使用单克隆抗体确定。 超微结构 抗原在链球菌上的位置将使用 免疫电子显微镜。 第三个目的是隔离结构 来自这些细菌的HBP基因并确定核苷酸序列 进一步表征这种毒力因子。 第四个目标是 使用插入生成链球菌缺乏HBP的菌株 诱变，并将它们与均匀的父母菌株进行比较以证明 该蛋白的致病特性。 从 本应用程序中提出的实验将进一步 了解人类中所有与感染相关的肾介质的理解，可能 最终允许开发安全的链球菌疫苗 防止感染而没有组织损伤的风险。