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| Author(s): |
Thais de Cassia Negrini
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba |
| Defense date: | 2008-02-22 |
| Examining board members: |
Renata de Oliveira Mattos Graner;
Denise Madalena Palomari Spolidorio;
José Francisco Höfling
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| Advisor: | Edgard Graner; Renata de Oliveira Mattos Graner |
| Abstract | |
Streptococcus mutans are the main pathogens of dental caries. Antigens (Ags) secreted or associated with the bacterial cell surface may influence in the susceptibility to infection by S. mutans. These include the virulence protein Glucan-binding protein B (GbpB). Recent studies indicate that the expression of surface antigens recognized by phagocytes is controlled by systems of two components (TCS, Two Component System) in several streptococci species. The objective of this project was to investigate the effect of variations in the production of GbpB and the influence of the two-component systems (TCS), Cov and Vic, in the pattern of internalization of S. mutans by macrophages from mice under the absence of opsonins. Thus, levels of bacterial internalization by macrophages were evaluated in 10 different clinical strains of S. mutans previously characterized regarding the patterns of GbpB production, and in four knockout mutants of the TCS, two covR- mutants and two vicH- mutants. To this purpose, murine macrophages of the lineage Balb/c were cultured in RPMI medium and exposed to different strains of S. mutans. The levels of macrophages with internalized bacteria were determined with the help of an optical microscope. Patterns of bacteria internalization were analyzed in similar assays using macrophages previously treated with inhibitors of specific mechanisms of internalization (phagocytosis, macropinocytosis and endocytosis mediated by clathrin) and by electron transmission microscopy (ETM) analyses. There was no statistically significant difference in the efficiency of internalization between the strains that differed regarding production of GbpB. The mutants vicH- was significantly less internalized when compared with the respective wild type strains and mutants covR-. Analyses with specific inhibitors and ETM indicated that S. mutans can be internalized by phagocytosis, macropinocytosis and endocytosis mediated by clathrin. These data may help to understand the mechanisms by which S. mutans is internalized and processed by antigen-presenting cells and the factors affecting patterns of adaptative immune response to these bacteria (AU) | |