We have recently demonstrated that human peripheral blood lymphocytes (PBL) contain a population of large granular lymphocytes which preferentially lyse HeLa cells infected with Shigella flexneri. In this proposal we will investigate the effector cells involved in this preferential killing of bacteria- infected cells, and the mechanism by which Shigella pretreatment leads to increased susceptibility to cytotoxic effector cell activity. The surface phenotype of cytotoxic effector cells present in human PBL, and in mouse spleen and gut associated lymphoid tissues (GALT) will be determined using different well characterized monoclonal antisera. The specificity of these cytotoxic effector cells will be addressed by using cold target inhibition and monolayer absorption types of experiments using different tumor cells and virus-infected cells. The susceptibility of bacteria-infected cells to soluble factors produced by natural killer (NK) cells will be investigated and compared to other tumor cells and virus-infected cells. To investigate the mechanism(s) by which Shigella renders a cell susceptible to natural cytotoxic effector cell activity, we will investigate the following: 1) requirement for bacteria invasion, 2) requirement for cell membrane alterations and/or presence of bacteria antigens, and 3) require- ment for cell versus bacteria metabolism. Using recombinant DNA methodology, we will isolate the bacterial gene(s) and define the gene product(s) important for demonstrating natural cytotoxic activity. Finally, we will investigate whether natural cytotoxic activity can be demonstrated against cells infected with other facultative intracellular bacteria. Results from these studies will not only contribute significantly to our understanding of immunity to facultative intracellular bacteria but will also provide the information necessary for establishing in vitro models for immune recognition of bacteria-infected cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023731-02
Application #
3136070
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1988-02-01
Project End
1991-01-31
Budget Start
1989-02-01
Budget End
1990-01-31
Support Year
2
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Type
Schools of Medicine
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
Arnold, J W; Niesel, D W; Annable, C R et al. (1993) Tumor necrosis factor-alpha mediates the early pathology in Salmonella infection of the gastrointestinal tract. Microb Pathog 14:217-27
Arnold, J W; Klimpel, G R; Niesel, D W (1993) Tumor necrosis factor (TNF alpha) regulates intestinal mucus production during salmonellosis. Cell Immunol 151:336-44
Brooks, E G; Wirt, D P; Klimpel, G R et al. (1993) In vivo and in vitro suppression of T-cell receptor alpha/beta CD4- CD8- T lymphocytes by cyclosporine A. Clin Immunol Immunopathol 67:224-31
Ramarathinam, L; Shaban, R A; Niesel, D W et al. (1991) Interferon gamma (IFN-gamma) production by gut-associated lymphoid tissue and spleen following oral Salmonella typhimurium challenge. Microb Pathog 11:347-56
Brooks, E G; Wirt, D P; Goldblum, R M et al. (1990) Double-negative (CD4- CD8-) T cells with an alpha/beta T cell receptor. Non-MHC-restricted cytolytic activity and lymphokine production. J Immunol 144:4507-12
Hess, C B; Niesel, D W; Holmgren, J et al. (1990) Interferon production by Shigella flexneri-infected fibroblasts depends upon intracellular bacterial metabolism. Infect Immun 58:399-405
Hess, C B; Niesel, D W; Klimpel, G R (1989) The induction of interferon production in fibroblasts by invasive bacteria: a comparison of Salmonella and Shigella species. Microb Pathog 7:111-20
Klimpel, G R; Niesel, D W; Asuncion, M et al. (1988) Natural killer cell activation and interferon production by peripheral blood lymphocytes after exposure to bacteria. Infect Immun 56:1436-41