STREPTOCOCCUS MUTANS INTERACTION WITH ANIMAL TISSUES

变形链球菌与动物组织的相互作用

• 批准号: 3219584
• 负责人: MURRAY W STINSON
• 金额: $ 15.94万
• 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 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219584
• 关键词: Streptococcus mutans; active immunization; affinity chromatography; bacterial antigens; bacterial toxicology; brain cell; cell membrane; cell wall; cross immunity; dental caries vaccine; electrofocusing; electron microscopy; gel electrophoresis; gel filtration chromatography; heart cell; human tissue; immunoelectrophoresis; immunofluorescence technique; immunoglobulin G; immunologic techniques; immunotoxicity; kidney cell; liver cells; microorganism culture; muscle cells; passive immunization; serology /serodiagnosis; serotyping; skin; surface antigens

The objectives of the project are to identify and characterize Streptococcus mutans cell components that may exert detrimental effects on host tissue through direct interactions or by immunological mechanisms and to establish methodologies for the evaluation of future vaccines. One phase of the study will be concerned with antigens that are shared by S. mutant and human and monkey heart and kidney tissue (CR-antigens). CR-antigens (CR-50 and CR-24) will be extracted from S. mutans 10449 and purified using a variety of methods, including: gel filtration, isoelectric focusing, polyacrylamide disc gel electrophoresis and affinity chromatography. Groups of New Zealand white rabbits will be immunized with the purified bacterial antigens using intravenous and intradermal injections. Rabbit sera will be tested for antibodies to CR-antigens by indirect immunofluorescence (IIF), immunoelectrophoresis and indirect radioimmunoassay (IRIA). The effects of streptococcal induced tissue-reactive antibodies will be studied in actively and passively immunized rabbits. Resulting tissue pathology will be ascertained using both light and electron microscopy. Immunofluorescence, immunoenzyme, immunoferritin and conventional histological stains will be employed. The CR-antigen of animal tissue will be solubilized, purified, and biochemically and immunologically characterized. In a second study, other antigens of S. mutants 10449 cells will be tested for direct binding activities for heart and kidney tissue using IIF, IRIA and direct binding assays using radiolabeled bacterial components. The tissue-binding components (TB-antigens) and the tissue receptor will be isolated, purified, and characterized using the methods described above. The effects of the streptococcal TB-antigens (TB65, TB35) in vivo will be studied by injection of purified material into rabbits or by perfusion into isolated kidneys and hearts. Tissue pathology will be determined with the methods listed above.

该项目的目标是确定和表征 可能对人体产生有害影响的变形链球菌细胞成分 宿主组织通过直接相互作用或通过免疫机制 建立评估未来疫苗的方法。 一 研究的第二阶段将关注 S. 共享的抗原。 突变体以及人类和猴子的心脏和肾脏组织（CR 抗原）。 CR-抗原（CR-50 和 CR-24）将从变形链球菌 10449 中提取，并 使用多种方法纯化，包括：凝胶过滤、 等电聚焦、聚丙烯酰胺圆盘凝胶电泳和亲和力 色谱法。 新西兰白兔群体将接受免疫接种 使用静脉内和皮内注射纯化细菌抗原 注射。 将通过以下方式测试兔血清中的 CR 抗原抗体： 间接免疫荧光（IIF）、免疫电泳和间接 放射免疫分析（IRIA）。 链球菌引起的影响 组织反应性抗体将被主动和被动地研究 免疫过的兔子。 由此产生的组织病理学将使用确定 光学显微镜和电子显微镜。 免疫荧光、免疫酶、 将使用免疫铁蛋白和常规组织学染色。 这 动物组织的CR抗原将被溶解、纯化，并 具有生化和免疫学特征。 在第二项研究中，其他 S.突变体10449细胞的抗原将被测试直接结合 使用 IIF、IRIA 和直接结合对心脏和肾脏组织进行活性 使用放射性标记的细菌成分进行测定。 组织结合 成分（结核抗原）和组织受体将被分离， 使用上述方法进行纯化和表征。 效果 体内链球菌结核抗原（TB65、TB35）的研究将通过 将纯化的材料注射到兔子体内或通过灌注到隔离的体内 肾脏和心脏。 组织病理学将用以下方法确定 上面列出的。