ROLE OF MAJOR OUTER MEMBRANE PROTEINS IN IMMUNE RESPONSE TO SALMONELLA

革兰氏阴性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双层中（掩埋的表位）。 本提案涉及 使用单克隆抗体（MAbs），已经提出了对这两个 表位的类别，以阐明主要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 为了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， 评估体液免疫中表面和埋藏表位的相对重要性， 对沙门氏菌感染的免疫反应。 抗原保守性 沙门氏菌孔蛋白将通过筛选约100 沙门氏菌血清型通过针对一组抗-（S。 鼠伤寒沙门氏菌）OmpD和OmpC MAb。 孔蛋白的免疫化学结构 表位将通过抗孔蛋白单克隆抗体的Western免疫印迹分析， 溴化氰（CNBr）取代孔蛋白单体，并通过氨基酸和N- CNBr肽的末端序列分析。 最后，相对 表位和埋藏表位在体液免疫应答中重要性 感染将通过检测识别这两种类型的单克隆抗体来研究 被动保护小鼠免受感染的能力 由S.鼠伤寒。 在拟定的研究中，鼠沙门氏菌病 它具有许多肠热病的特征， 建立了一个研究沙门氏菌致人伤寒的系统。伤寒。 的 该项目的长期目标是阐明 模拟某些表位的主要OM蛋白或肽在 这些蛋白质作为抗S.伤寒感染。 因为所有的克- 阴性菌可能含有相似的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 该项目还旨在培训少数民族学生， 参与这个研究项目的各个阶段。 学生将复习 文献，制定研究思路，设计和开展实验 使用现代生物医学研究技术，并获得写作经验 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.