ROLE OF MAJOR OUTER MEMBRANE PROTEINS IN IMMUNE RESPONSE TO SALMONELLA

革兰氏阴性细菌的外膜蛋白含有表位 暴露在细胞表面的表位(表面表位)，以及 被掩埋在OM双层(掩埋表位)中。这项建议涉及的是 使用已经提出的针对这两种病毒的单抗(MAb) 表位分类，阐明主要OM蛋白在免疫中的作用 对沙门氏菌感染的反应。该项目的具体目标是 以确定孔蛋白表位在属内的保守程度 沙门氏菌，研究保守结构域的免疫化学结构，以及 体液表面表位和埋入表位的相对重要性评价 对沙门氏菌感染的免疫反应。猪瘟病毒的抗原性保守 沙门氏菌孔菌素将通过筛查约100 用免疫印迹法检测沙门氏菌的血清型。 小鼠)OmpD和OMPC单抗。孔蛋白的免疫化学结构 将通过免疫印迹技术分析抗孔蛋白单抗的表位 溴化氰(CNBr)对孔蛋白单体的消化，以及氨基酸和N- CNBr多肽的末端序列分析。最后，相对的 表面和埋藏表位在体液免疫应答中的重要性 将通过检测识别这两个类别的单抗来研究感染情况 它们能够被动保护小鼠免受感染的表位 鼠伤寒沙门氏菌。在拟议的调查中，小鼠沙门氏菌病 它具有许多肠热病的特征，将被用作模型 伤寒沙门氏菌引起的人类伤寒研究系统。这个 这个项目的长期目标是阐明 主要OM蛋白或模拟某些表位的多肽在农业生产中的应用 这些蛋白质被用作对抗伤寒沙门氏菌感染的疫苗。因为所有的奶奶- 阴性细菌可能含有类似的OM蛋白结构， 拟议的研究与革兰氏阴性细菌的发病机制有关 将军。该项目还旨在培训将 参与本研究项目的所有阶段。学生们将复习 查阅文献，拟定研究思路，设计并开展实验 使用现代生物医学研究技术，并获得写作经验 以及在区域和国家会议上提交科学论文。

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.