STREPTOCOCCUS MUTANS INTERACTION WITH ANIMAL TISSUES

变形链球菌与动物组织的相互作用

• 批准号: 3219586
• 负责人: MURRAY W STINSON
• 金额: $ 15.71万
• 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 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219586
• 关键词: Streptococcus mutans; active immunization; adhesin; bacterial antigens; bacterial toxicology; basement membrane; cell membrane; cell wall; cross immunity; crosslink; dental caries vaccine; electron microscopy; human tissue; immunoelectrophoresis; immunofluorescence technique; immunoglobulin G; immunologic techniques; immunotoxicity; kidney cell; laboratory rabbit; laminin; membrane activity; microorganism culture; nephritis; oral bacteria; passive immunization; radiotracer; serology /serodiagnosis; skin; surface antigens; vascular endothelium; vascular endothelium permeability

Streptococcus mutans produces several components that bind to basement membranes and endothelial cells of kidney in vitro. These adhesins are secreted into the environment during growth and are also associated with the bacterial cell surface. It is believed that these substances may leach naturally into blood vessels of the oral cavity, or enter as a consequence of trauma to oral tissues or by parenteral immunization. In rabbits, the interaction of S. mutans components with kidney results in a severe nephritis that shows many histological, immunopathological, and serological features of streptococcus- associated nephritides in man. Pathogenesis may result from either direct toxicity of the tissue-bound adhesin or in situ activation of the complement cascade by the regular or alternative pathways. The long range goals of this project are to: a) understand the mechanisms of adhesin binding and pathogenesis; b) to improve the safety of prospective carries vaccines by assuring exclusion of these hazardous factors; c) devise prophylactic or therapeutic measures for streptococcus- associated nephritis. Specifically, the nature of the interactions between known S. mutans adhesions (LTA and proteins P57, P48, P40 and P30) and kidney components in vitro will be defined. The kidney-binding pattern of each purified adhesin will be characterized by immunofluorescence and immunoenzyme stains in light microscopy. Binding specificities of radiolabeled adhesins and the identities of the corresponding kidney components will be determined using isotopic dilution techniques and competitive inhibition assays using purified tissue components; e.g., laminin and heparan sulfate. Photoactivated crosslinking reagents will be used to covalently couple streptococcal protein to its kidney receptor which will then be isolated and characterized. The in vivo binding activities and pathogenic properties of purified S. mutans adhesins will be studied in a rabbit experimental model. The effects of infusion and perfusion of kidneys and intravenous injections of rabbits will be evaluated by microscopic analyses or renal biopsies using immunofluorescence, immunoenzyme and histological stains. Electron microscopy with immunogold labelling will also be used. In vivo experiments will also reveal the modulating effects of antibodies, complement and other serum components on adhesin binding. Finally, bacteria in dental plaque will be tested for both cell wall-associated and extracellular adhesins using immunofluorescence and electron microscopy.

链球菌突变产生的几个组成部分结合 体外肾脏的基底膜和内皮细胞。 这些粘合剂在生长过程中分泌到环境中 并且也与细菌细胞表面有关。 这是 相信这些物质可能会自然浸入血液中 口腔的血管，或因创伤而进入 口腔组织或通过肠胃外免疫。 在兔子中 链球菌成分与肾脏的相互作用导致A 严重的肾炎显示许多组织学， 免疫病理学和链球菌的血清学特征 相关的肾植物。 发病机理可能是由 组织结合的粘附素的直接毒性或原位 定期激活补充级联反应 替代途径。 该项目的远距离目标是 to：a）了解粘附蛋白结合的机制和 发病; b）提高潜在携带的安全性 通过确保排除这些危险因素，疫苗； c） 设计预防或治疗措施 相关的肾炎。 具体而言，互动的本质 在已知的链球菌粘连（LTA和蛋白质p57，p48， 将定义P40和P30）和肾脏成分。 这 每种纯化的粘附素的肾脏结合模式将是 以免疫荧光和免疫酶染色为特征 在光学显微镜中。 放射性标记粘合剂的结合特异性 相应的肾脏成分的身份将是 使用同位素稀释技术和竞争性确定 使用纯化的组织成分抑制测定；例如，层粘连蛋白 和硫酸乙酰肝素。 光活化的交联试剂将是 用于共价将链球菌蛋白与肾脏搭配 然后将被隔离和表征的受体。 in 纯化的S.体内结合活性和致病特性 突变蛋白将在兔实验模型中进行研究。 肾脏和静脉注射的灌注的影响 兔子注射将通过微观分析或 使用免疫荧光，免疫酶和 组织学染色。 电子显微镜与免疫元 标签也将使用。 体内实验也将揭示 抗体，补体和其他的调节作用 粘附素结合上的血清成分。 最后，牙齿中的细菌 将对两种细胞壁相关和 使用免疫荧光和电子的细胞外粘合剂 显微镜。