The objectives of the proposed enrichment plan are: (1) to provide opportunities for minority science students to become acquainted with and to participate in biomedical research in order to motivate them to pursue graduate studies in the biomedical sciences, and (2) to provide opportunities for science faculty to upgrade their research skills and be involved in active sponsored research and student training. The proposed activities include: (a) faculty and student participation in seminars and workshops conducted by visiting scientists at Alabama State University campus, (b) faculty participation in workshops, extramural research, and """"""""retooling"""""""" experiences at off-campus research institutions, (c) students' participation in extramural biomedical research during the summer at major universities in the United States, (d) students taking BIO 350, Introduction to Biomedical Research Techniques at Alabama State University, (e) faculty and student participation in Alabama State University's Biomedical Science Club activities and the College of Arts and Science's annual research symposium, and (f) attendance (by faculty and students) and presentation of research papers at national scientific meetings. We believe these activities will contribute to an improved intellectual and scientific environment in natural sciences which will motivate more science faculty to get actively involved in research, and more students to pursue graduate studies in biomedical sciences. Grant=S06GM082190006 Salmonella typhimurium is a gram-negative pathogen that primarily evokes gastroenteritis in man, but causes typhoid fever-like disease in mice. The outer membrane (OM) of S. typhimurium and other gram-negative bacteria contains lipopolysaccharide (LPS) and a family of major proteins called, porins. These OM components are the primary targets for immune recognition by the host during bacterial infections. This project will sequence the gene that encodes OmpD porin, and use monoclonal antibodies (MAbs) to determine the molecular specificity of the humoral immune response to infection by S. typhimurium.
The specific aims of the project are to map the antigenic regions on the OmpD porin of Salmonella, to clone and sequence the ompD gene, to identify the dominant and immunoprotective porin and porin-LPS epitopes, and to determine the effect of LPS structure on the recognition of these epitopes during bacterial infections. The antigenic regions of OmpD will be mapped by Western immunoblot reactivity of cyanogen bromide-generated porin peptides with a panel of existing anti-OmpD MAbs. Identification of dominant (and immunoprotective) epitopes will involve the analysis of polyclonal immune sera by binding competition (enzyme-linked immunosorbent assay and Western blots) with a panel of existing anti- Salmonella MAbs. These MAbs are either specific for the surface epitopes of OmpD or OmpC porin, or for the porin-LPS complex, and they either prolong the survival of mice or confer significant protection against endotoxemia and/or bacteremia in experimental murine salmonellosis. The effect of LPS structure on the specificity of anti-porin and anti-LPS antibodies that arise during infection will be studied using smooth and rough variants of S. typhimurium. The long-term goal of this project is to elucidate the potential application of immunodominant porin epitopes for development of Salmonella-specific diagnostic probes and vaccines, and for insertion and expression of foreign genes. 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.
