ROLE OF MAJOR OUTER MEMBRANE PROTEINS IN IMMUNE RESPONSE TO SALMONELLA

主要外膜蛋白在沙门氏菌免疫反应中的作用

• 批准号: 6240269
• 负责人: SHIVA P SINGH
• 金额: $ 21.81万
• 依托单位: ALABAMA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6240269
• 关键词: SDS polyacrylamide gel electrophoresis; Salmonella; Salmonella infections; Salmonella vaccines; enzyme linked immunosorbent assay; genetic mapping; gram negative bacteria; humoral immunity; immunochemistry; immunoprecipitation; laboratory mouse; membrane proteins; molecular cloning; monoclonal antibody; protein sequence; protein structure function; western blottings

The outer membrane (OM) proteins in gram-negative bacteria contain epitopes that are exposed on the cell surface (surface epitopes), and epitopes that are buried in the OM bilayer (buried epitopes). This proposal deals with the use of monoclonal antibodies (MAbs), already raised against the two classes of epitopes, to elucidate the role of major OM proteins in immune response to infection by Salmonella. The specific aims of the project are to determine the extent of conservation of porin epitopes within the genus Salmonella, study the immunochemical structure of conserved domains, and evaluate the relative importance of surface and buried epitopes in humoral immune response to infection by Salmonella. the antigenic conservation of Salmonella porins will be determined by screening approximately 100 Salmonella serotypes by Western immunoblot against a panel of anti-(S. typhimurium) OmpD and OmpC MAbs. The immunochemical structure of the porin epitopes will be analyzed by Western immunoblot of antiporin MAbs against cyanogen bromide (CNBr) digests of porin monomers, and by amino acid and N- terminal sequence analyses of CNBr peptides. Finally, the relative importance of surface and buried epitopes in humoral immune response to infection will be studied by testing MAbs which recognize the two classes of epitopes for their ability to passively protect mice against infection by S. typhimurium. In the proposed investigation, murine salmonellosis which has many of the hallmarks of enteric fever will be used as a model system for the study of human typhoid disease induced by S. typhi. The long-term objective of this project is to elucidate the potential application of major OM proteins or peptides mimicking certain epitopes on these proteins as vaccines against S. typhi infections. Since all gram- negative bacteria probably contain similar OM protein structures, the proposed research is relevant to gram-negative bacterial pathogenesis in general. The project is also designed to train minority students who will participate in all phases of this research project. Students will review literature, formulate research ideas, design and carry out experiments using modern biomedical research techniques, and gain experience in writing and presentation of scientific papers at regional and national meetings.

革兰氏阴性细菌中的外膜（OM）蛋白包含表位 暴露在细胞表面（表面表位）和表位的表面 被埋葬在OM双层（埋入的表位）中。 该提议处理 单克隆抗体（mAb）的使用，已经针对两种抗体 表位类别，以阐明主要OM蛋白在免疫中的作用 沙门氏菌对感染的反应。 该项目的具体目的是 确定属内孔蛋白表位的保守程度 沙门氏菌，研究保守域的免疫化学结构， 评估表面和掩埋表位的相对重要性 沙门氏菌对感染的免疫反应。 抗原保护 沙门氏菌将通过筛查约100 西方免疫印迹的沙门氏菌血清型针对一组抗小组（s。 鼠伤寒和OMPC mAb。 孔蛋白的免疫化学结构 西方免疫印迹将对抗磷灰蛋白mab的免疫印迹分析表位 Porin单体的氰基溴化物（CNBR）摘要，以及氨基酸和N- CNBR肽的末端序列分析。 最后，亲戚 表面和掩埋表位的重要性在体液免疫反应中 将通过测试MAB来研究感染，以识别这两个类别 表位的能力被动保护小鼠免受感染的能力 由鼠伤寒链球菌。 在拟议的调查中，鼠沙门氏菌病 具有许多肠发烧的标志将用作模型 链球菌诱导的人伤寒疾病的研究系统。 这 该项目的长期目标是阐明潜力 在模仿某些表位上的主要OM蛋白或肽的应用 这些蛋白质是针对链球菌感染的疫苗。 由于所有革兰氏 阴性细菌可能包含相似的OM蛋白结构， 提出的研究与革兰氏阴性细菌发病机理有关 一般的。 该项目还旨在培训少数民族学生 参与该研究项目的所有阶段。 学生将审查 文学，制定研究思想，设计和执行实验 使用现代的生物医学研究技术，并获得写作经验 以及在区域和国家会议上的科学论文的介绍。