Ruchele Dias Nogueira

Characterization of Salivary Immunoglobulin A Responses in Children Heavily Exposed to the Oral Bacterium Streptococcus mutans: Influence of Specific Antigen Recognition in Infection

ABSTRACT The initial infection of children by Streptococcus mutans, the main pathogen of dental caries, depends on the ability of S. mutans to adhere and accumulate on tooth surfaces. These processes involve the adhesin antigen I/II (AgI/II), glucosyltransferases (GTF) and glucan-binding protein B (GbpB), each a target for anticaries vaccines. The salivary immunoglobulin A (IgA) antibody responses to S. mutans antigens (Ags) were characterized in 21 pairs of 5- to 13-month-old children. Pairs were constructed with one early S. mutans-infected and one noninfected child matched by age, racial background, number of teeth, and salivary levels of IgA. Specific salivary IgA antibody response and S. mutans infection levels were then measured during a 1-year follow-up. Robust responses to S. mutans were detected from 6 months of age. Salivary IgA antibody to AgI/II and GTF was commonly detected in salivas of all 42 children. However, GbpB-specific IgA antibody was seldom detected in the subset of infected children (38.1% at baseline). In contrast, most of the subset of noninfected children (76.2%) showed GbpB-reactive IgA antibody during the same period. Frequencies of GbpB responses increased with age, but differences in intensities of GbpB-IgA antibody reactions were sustained between the subsets. At baseline, GbpB-reactive IgA antibody accounted for at least half of the total salivary IgA S. mutans-reactive antibody in 33.3 and 9.5% of noninfected and infected children, respectively. This study provides evidence that a robust natural response to S. mutans Ags can be achieved by 1 year of age and that IgA antibody specificities may be critical in modulating initial S. mutans infection.

Prospective study of potential sources of Streptococcus mutans transmission in nursery school children.

Transmission of Streptococcus mutans, a major dental caries pathogen, occurs mainly during the first 2.5 years of age. Children appear to acquire S. mutans mostly from their mothers, but few studies have investigated non-familial sources of S. mutans transmission. This study prospectively analysed initial S. mutans oral colonization in 119 children from nursery schools during a 1.5-year period and tracked the transmission from child to child, day-care caregiver to child and mother to child. Children were examined at baseline, when they were 5-13 months of age, and at 6-month intervals for determination of oral levels of S. mutans and development of caries lesions. Levels of S. mutans were also determined in caregivers and mothers. A total of 1392 S. mutans isolates (obtained from children, caregivers and mothers) were genotyped by arbitrarily primed PCR and chromosomal RFLP. Overall, 40.3 % of children were detectably colonized during the study, and levels of S. mutans were significantly associated with the development of caries lesions. Identical S. mutans genotypes were found in four nursery cohorts. No familial relationship existed in three of these cohorts, indicating horizontal transmission. Despite high oral levels of S. mutans identified in most of the caregivers, none of their genotypes matched those identified in the respective children. Only 50 % of children with high levels of S. mutans carried genotypes identified in their mothers. The results support previous evidence indicating that non-familial sources of S. mutans transmission exist, and indicate that this bacterium may be transmitted horizontally between children during the initial phases of S. mutans colonization in nursery environments.

Mutans Streptococcal Infection Induces Salivary Antibody to Virulence Proteins and Associated Functional Domains

ABSTRACT The interplay between mucosal immune responses to natural exposure to mutans streptococci and the incorporation and accumulation of these cariogenic microorganisms in oral biofilms is unclear. An initial approach to explore this question would be to assess the native secretory immunity emerging as a consequence of Streptococcus mutans infection. To this end, we analyzed salivary immunoglobulin A (IgA) antibody to mutans streptococcal glucosyltransferase (Gtf) and glucan binding protein B (GbpB) and to domains associated with enzyme function and major histocompatibility complex (MHC) class II binding in two experiments. Salivas were collected from approximately 45-day-old Sprague-Dawley rats, which were then infected with S. mutans SJ32. Infection was verified and allowed to continue for 2 to 2.5 months. Salivas were again collected following the infection period. Pre- and postinfection salivas were then analyzed for IgA antibody activity using peptide- or protein-coated microsphere Luminex technology. S. mutans infection induced significant levels of salivary IgA antibody to Gtf (P < 0.002) and GbpB (P < 0.001) in both experiments, although the levels were usually far lower than the levels achieved when mucosal immunization is used. Significantly (P < 0.035 to P < 0.001) elevated levels of postinfection salivary IgA antibody to 6/10 Gtf peptides associated with either enzyme function or MHC binding were detected. The postinfection levels of antibody to two GbpB peptides in the N-terminal region of the six GbpB peptides assayed were also elevated (P < 0.031 and P < 0.001). Interestingly, the patterns of the rodent response to GbpB peptides were similar to the patterns seen in salivas from young children during their initial exposure to S. mutans. Thus, the presence of a detectable postinfection salivary IgA response to mutans streptococcal virulence-associated components, coupled with the correspondence between rat and human mucosal immune responsiveness to naturally presented Gtf and GbpB epitopes, suggests that the rat may be a useful model for defining mucosal responses that could be expected in humans. Under controlled infection conditions, such a model could prove to be helpful for unraveling relationships between the host response and oral biofilm development.

Age-Specific Salivary Immunoglobulin A Response to Streptococcus mutans GbpB

ABSTRACT In a follow-up study of children infected with Streptococcus mutans at an early age (children previously shown to respond poorly to S. mutans GbpB), there was a delay in their immune response, rather than a complete inability to respond to this antigen. Epitopes in the N-terminal third of GbpB were identified as targets for naturally induced immunoglobulin A antibody in children at an early age.

Galectin-9 as an important marker in the differential diagnosis between oral squamous cell carcinoma, oral leukoplakia and oral lichen planus

OBJECTIVES To evaluate the expression of Galectins (Gal) 1, 3 and 9, Metalloproteinase 3 (MMP-3) and mast cell density in oral lesions of patients with potentially malignant disorders (PMD) and oral squamous cell carcinomas (OSCC) by comparison with the controls. STUDY DESIGN We selected 40 cases of PMD, 40 OSCC and 13 with normal histopathological profile. Immunohistochemistry was performed for Gal-1, Gal-3, Gal-9 and MMP-3. RESULTS Gal-9 was significantly higher in patients with OSCC than in others groups (p < 0.001). Gal-1 expression was significantly lower in patients with leukoplakia than those with OSCC and controls (p = 0.0001). Gal-3 was significantly lower in patients with OSCC than those with leukoplakia (p = 0.03). MMP-3 was lower in patients with leukoplakia in comparison with the lichen planus group (p = 0.013). CONCLUSION The increased expression of Gal-9 may be helpful to differentiate of OSCC from other oral cavity lesions.

Epitopes shared among pioneer oral flora and Streptococcus mutans GbpB

The establishment of microorganisms in emerging oral biofilms of humans is likely to be modulated by a constellation of infant, maternal, and microbial factors. We have shown that pioneer microbiota on epithelial surfaces (e.g., Streptococcus mitis, Streptococcus salivarius) and initially erupting dental surfaces (e.g., Streptococcus sanguinis, Streptococcus gordonii, and Streptococcus oralis) induce mucosal SIgA antibody in saliva. The resulting immune responses may modulate subsequent colonization of bacteria that later join the oral biofilm (e.g., cariogenic Streptococcus mutans), a speculation supported by the observation that (a) several S. mutans virulence components share MHC Class II binding peptides with pioneer protein homologues, (b) cross-reactive antibody among homologues can be demonstrated and (c) initial S. mutans colonization can apparently be delayed when some of these responses are present a priori.

Levels of Pro and Anti-inflammatory Citokynes and C-Reactive Protein in Patients with Chronic Periodontitis Submitted to Nonsurgical Periodontal Treatment

Aim to compare the levels of IFN-γ, TGF-β and C-reactive protein (CRP) in healthy patients (HP) and chronic periodontitis patients (CP) before and seven days after the last session of Non-Surgical Periodontal Treatment (NSPT). Materials and Methods 40 subjects were divided into two groups: healthy (n= 20), and with chronic periodontitis (n = 20). Serum and gingival crevicular fluid (GCF) were collected from each patient and quantified for IFN-γ, TGF-β and CRP using the enzyme-linked immunosorbent assay (ELISA). Results IFN-γ was found to be higher in the GCF of the CP group before NSPT in relation to the HP group (p<0.05), and it had significant higher levels after seven days of NSPT (p<0.05). The levels of TGF-β in the GCF of CP patients before NSPT were significantly higher when compared to HP (p<0.05), but they decreased after seven days of NSPT (p>0.05). Serum CRP levels did not show statistical difference between CP and HP before or after NSPT. Conclusion Therefore, our results demonstrated for the first time that NSPT causes early exacerbation of the immune response at the local level represented by increased levels of IFN-γ and decreased levels of TGF-β in the gingival crevicular fluid after seven days of treatment.