The long-term goal of the College of Veterinary Medicine (CVM) Flow Cytometry Core is to have a flow cytometry facility complete with analytical instruments and one instrument for cell-sorting. This equipment grant proposal seeks to expand its capacity for cell-sorting to obtain lymphocyte and leukocyte subsets to >95% cell purity. The Sony SH800 Cell Sorter, proposed herein, will fulfill this shortcoming of having a user-friendly system for high- speed cell-sorting for use by a diverse, but highly interactive grou of mucosal immunologists with a strong focus in one-health investigations. With the recent expansion of mucosal immunologists in CVM, the College of Medicine (CoM), and the College of Dentistry (CoD) at the University of Florida (UF), this group desires a cost- effective instrument not requiring a dedicated operator, and an instrument that can be supported by this core. Given that the greatest concentration of mucosal immunologists has been in CVM, the proposed Sony SH800 will be housed in CVM, but readily accessible to all major users within the three colleges as well as to other investigators within CVM having a need for cell-sorting for their health-related studies. With the availability of the Sony SH800, we envision it will immediately support, at the minimum, of 8 major users and one minor user. Cell- sorting will be used to fulfill study objectives in currently NIH-funded programs for development of mucosal vaccines for brucellosis, anthrax, botulism, enteric pathogens, plague, tuberculosis, and norovirus. In addition, this instrument will facilitate studies understanding the basic mechanisms of aryl hydrocarbon receptor signaling for the development of mucosal Th17 cells and regulatory T cells in colitis; studies understanding the TCR usage in animals and humans with Sjgren's Syndrome; studies understanding the influences of the gut microbiota upon intestinal epithelial and lymphoid cells in the normal and inflamed gut; and studies understanding how TLR signaling via the oral mucosa impacts lymphocyte responses to periodontal disease and oral neoplasms. Included in this proposal is the Baker Biological Containment Cabinet specifically designed to house the Sony SH800 to contain aerosols produced during cell-sorting. Since this collective group of investigators work with infected animal and human tissue samples, this barrier provides a needed additional level of safety for both the operators and samples processed. In summary, having the capacity for high-speed cell-sorting at a reasonable cost will further the current collaborations amongst these mucosal immunologists, help them meet their study objectives for ongoing NIH-funded research, and expand their ongoing effectiveness in infectious and autoimmune disease research.
The mucosal immunology group at the University of Florida has strengths in developing vaccines and therapeutics for infectious and autoimmune diseases. The goal of this equipment grant is to acquire the Sony SH800 cell- sorter, which in turn will facilitate obtaining defined lymphocyte or leukocyte subsets to >95% purity. The Sony SH800 will enhance or expand the capabilities of this group to study a variety of different mucosal diseases supported by NIH research grant awards.