RESUMEN
We have previously shown that one of the minimal active regions of statherin, a human salivary protein, for binding to Fusobacterium nucleatum is a YQPVPE amino acid sequence. In this study, we identified the FomA protein of F. nucleatum, which is responsible for binding to the statherin-derived YQPVPE peptide. Overlay analysis showed that a 40-kDa protein of the F. nucleatum cell envelope (40-kDa CE) specifically bound to the YQPVPE peptide. The equilibrium association constant between the affinity-purified 40-kDa CE and the YQPVPE peptide was 4.30 x 10(6). Further, the purity and amino acid sequence analyses of the purified 40-kDa CE revealed approximately 98.7% (wt/wt) purity and a high degree of homology with FomA, a major porin protein of F. nucleatum. Thus, a FomA-deficient mutant failed to bind to the YQPVPE peptide. In addition, increased levels of a FomA-specific mucosal IgA antibody (Ab) and plasma IgG and IgA Abs were seen only in mice immunized nasally with cholera toxin (CT) and the purified 40-kDa FomA protein. Interestingly, saliva from mice that received FomA plus CT as a mucosal adjuvant nasally prevented in vitro binding of F. nucleatum to statherin-coated polyvinyl chloride plates. Taken together, these results suggest that induction of specific immunity to the 40-kDa FomA protein of F. nucleatum, which specifically binds to the statherin-derived peptide, may be an effective tool for preventing the formation of F. nucleatum biofilms in the oral cavity.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/inmunología , Proteínas de la Membrana Bacteriana Externa/metabolismo , Fusobacterium nucleatum/inmunología , Fusobacterium nucleatum/patogenicidad , Mapeo de Interacción de Proteínas , Proteínas y Péptidos Salivales/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos Antibacterianos/análisis , Anticuerpos Antibacterianos/sangre , Anticuerpos Antibacterianos/inmunología , Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/genética , Femenino , Fusobacterium nucleatum/genética , Eliminación de Gen , Humanos , Inmunoglobulina A/análisis , Inmunoglobulina A/sangre , Inmunoglobulina A/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Cinética , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Peso Molecular , Membrana Mucosa/inmunología , Unión ProteicaRESUMEN
Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralization. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralization in vitro. Sixty enamel specimens obtained from caries free human premolars were demineralised to produce caries-like lesions. Half of each lesion was covered with nail varnish as an untreated control. The specimens were then randomly divided into F and Sr+F treatment groups. The F group was exposed to remineralizing solutions (1.5mM CaCl(2), 0.9 mM KH(2)PO(4)) containing 1 ppm, 0.1 ppm or 0.05 ppm F. The Sr+F treatment group was exposed to the same solutions including 10 ppm Sr. After 2 weeks, lesion depth, mineral loss and percentage enamel remineralization were determined using transversal microradiography. There was a significant decrease in mineral loss in all groups (p<0.001). Lesion depth was significantly reduced for all groups (p<0.05) with the exception of group F. Remineralization was significantly affected by F concentration (p=0.000). The participation of Sr resulted in a significant enhancement of remineralization (p<0.001) with a synergistic effect of the Sr+F combination (p<0.01). It was concluded that while the remineralizing process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.
