IgA anti-Streptococcus mutans em crianças com e sem cárie dentária

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação das correlações existentes entre experiência de cárie, acúmulo de placa bacteriana e nível de Iga anti-streptococcus mutans na saliva de crianças com dentição mista

Pós-graduação em Odontologia Restauradora - ICT

Correlation among mutans streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva

Two-hundred and forty individuals were studied, divided into five groups as follows: caries-free children, children with caries, children with rampant caries, young adults with and without caries. Whole stimulated saliva was collected and all individuals were investigated for DMFT/dmft according to the WHO criteria and the simplified oral hygiene index (OHI-S). Quantitative analysis of the total aerobic flora and mutans streptococci in saliva was performed. Also, the level of salivary anti-S. mutans IgA was determined by ELISA. Children with rampant caries showed the highest OHI-S value. The highest total counts of microorganisms were found in the group of children with caries. No statistically significant differences were observed for salivary flow, OHI-S and microorganism counts between the groups of young adults. No correlation between mutans streptococci counts and anti-Streptococcus mutans IgA levels was observed in the studied groups. A correlation between increased anti-Streptococcus mutans IgA levels and caries-free status was observed among young adults but not among children.

Effects of probiotic bacteria on Candida presence and IgA anti-Candida in the oral cavity of elderly

Imbalance in the resident microbiota may promote the growth of opportunistic microorganisms, such as yeasts of Candida genus and the development of diseases, especially in aged people. This study evaluated whether the consumption of the probiotic Yakult LB® (Lactobacillus casei and Bifidobacterium breve) was able to influence on the specific immunological response against Candida and on the presence of these yeasts in the oral cavity of 42 healthy aged individuals. Saliva samples were collected before and after the probiotic use for 30 days, 3 times a week. The samples were plated in Dextrose Saboraud Agar with chloramphenicol, the colony-forming units (CFU/mL) were counted and the Candida species were identified. Anti-Candida IgA analysis was conducted using the ELISA technique. ANOVA and Student's t-test were used for normally distributed data and the Wilcoxon test was used for data with non-normal distribution (α=0.05). The results showed a statistically significant reduction (p<0.05) in Candida prevalence (from 92.9% to 85.7%), in CFU/mL counts of Candida and in the number of non-albicans species after consumption of the probiotic. Immunological analysis demonstrated a significant increase (p<0.05) in anti-Candida IgA levels. In conclusion, probiotic bacteria reduced Candida numbers in the oral cavity of the elderly and increased specific secretory immune response against these yeasts, suggesting its possible use in controlling oral candidosis.

Caries risk tests and salivary levels of immunoglobulins to Streptococcus mutans and Candida albicans in mouthbreathing syndrome patients

The aim of this study was to compare microbiological and salivary variables possibly related to caries risk in treated and untreated mouthbreathing syndrome (MBS) children and control children. Thirty control children, 30 mouthbreathers and 25 treated mouthbreathers were studied for the numbers of lactobacilli, mutans streptococci and yeasts in their saliva. Snyder's test, salivary flow and buffering capacity were also evaluated. Levels of immunoglobulins to Candida albicans and Streptococcus mutans in the saliva were quantified using ELISA. Considering the results obtained for the microbiological and salivary caries risk tests, no significant differences were observed among the proportions of patients with small/negative and high/moderate caries risk in the studied groups. The level of IgG to S. mutans was significantly higher in the treated MBS group in relation to MBS patients. on the other hand, the median anti-S. mutans IgM level was lower in the treated MBS patients than in the other groups. For the studied anti-Candida immunoglobulins, IgM level was significantly lower in the treated MBS group than in the other groups. No differences were observed for anti-S. mutans and anti-Candida IgA levels among the groups. The findings suggest that mouthbreathing cannot be considered a risk factor for dental caries. Copyright (C) 2003 S. KargerAG, Basel.

Presença de Candida nas mucosas vaginal e bucal e sua relação com IgA salivar

OBJETIVO: correlacionar a presença de leveduras do gênero Candida na cavidade bucal e vaginal de mulheres com e sem candidíase vulvovaginal (CVV) com os níveis de IgA secretora (IgAs) presentes na saliva. MÉTODOS: cinqüenta e uma mulheres foram incluídas; 13 apresentaram CVV e 38 formaram o Grupo Controle. de cada paciente, foram coletados 2,0 mL de saliva sem estimulação e secreção vaginal com o auxílio de swab, que foi imerso a seguir em 2,0 mL de solução fisiológica. As amostras foram semeadas em ágar Sabouraud dextrose com cloranfenicol para isolamento e contagem de colônias, e os isolados foram identificadas fenotipicamente. Na saliva de ambos os grupos foi quantificada IgA pela técnica ELISA. RESULTADOS: nas 13 pacientes com diagnóstico clínico e micológico de CVV, a média de unidades formadoras de colônias de Candida por mililitro de secreção vaginal (ufc/mL) foi de 52.723 e 23,8% dos pacientes apresentaram colonização na mucosa bucal com menor quantidade de ufc/mL (6.030). Os níveis de IgAs na saliva foram mais baixos no grupo com CVV (média de densidade: 0,3) quando comparados aos níveis de IgA do Grupo Controle (média de DO: 0,6). Onze pacientes (37%) do Grupo Controle apresentaram colonização por Candida na cavidade bucal, com média de ufc/mL mais baixa quando comparada ao grupo com CVV. O Grupo Controle também apresentou menor quantidade de ufc/mL (1.973) na cavidade vaginal quando comparado com o Grupo CVV (52.942). CONCLUSÕES: os resultados demonstraram que os pacientes com diagnóstico clínico de candidíase vulvovaginal apresentaram maior quantidade de Candida, tanto na cavidade vaginal quanto na bucal, e apresentaram menores níveis de IgA anti-Candida na saliva.

Production of monoclonal antibodies against Streptococcus mutans antigens

Several studies have been conducted in the last decades aiming to obtain an anti-canies vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, G8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG 2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

Differential antibody isotype expression to the major Paracoccidioides brasiliensis antigen in juvenile and adult form paracoccidioidomycosis

We investigated the relationship between antibody response to the major Paracoccidioides brasiliensis antigen, a 43-kDa glycoprotein, and the two paracoccidioidomycosis (PCM) clinical presentations, the juvenile and the adult forms. Total immunoglobulin G (IgG), IgG isotypes, and IgA anti-gp43 antibodies were determined by enzyme-linked immunosorbent assay in patients' sera. Juvenile PCM patients had higher (P =.003) IgG anti-gp43 levels than adult form patients. IgG1 subclass levels, however, were comparable between the two clinical forms. Patients with the juvenile form had higher (P <.001) IgG4, but lower(P =.03) IgG2 levels than patients with the adult form. The IgG4 isotype, regulated by interleukin 4, was found in all juvenile form patients but in only 12% of the adult form patients. In contrast, high levels of the IgG2 isotype, regulated by interferon-gamma, were found in 41% of the adult PCM patients, mainly those with a more benign disease, but in only 12% of the juvenile patients. IgG3 was either absent or detected at low levels. These results demonstrate, for the first time, specific IgG4 antibodies in the humoral immune response of patients with an endemic deep mycosis and suggest that the switch to the IgG subclasses in PCM is regulated by the patients' T-helper subset (Th-l or Th-2) dominant cytokine profile. A possible role for IgG4 in the immunopathogenesis of the juvenile, more severe form of the disease is discussed. Finally, IgA was found mainly in adult form patients, probably as a result of the chronic mucosal antigenic stimulation characteristic of this form. (C) Elsevier, Paris.

Relationship of abomasal histology and parasite-specific immunoglobulin A with the resistance to Haemonchus contortus infection in three breeds of sheep

This study was carried out to evaluate the relationship of abomasal inflammatory cells and parasite-specific immunoglobulin A (IgA) in mucus, with the resistance to Haemonchus contortus infection in three breeds of sheep naturally infected with gastrointestinal nematodes. The breeds were the native Santa Ines sheep, and the European Suffolk and Ile de France breeds. Mast cells, eosinophils and globule leucocytes were enumerated in abomasal mucosa. Eosinophils within the sub-mucosa also were counted separately. Histamine concentration was estimated in abomasal tissue samples. Enzyme-linked immunosorbent assay was carried out in mucus samples to determine the level of IgA anti-H. contortus third and fifth instar. There were no significant differences among group means of these variables (P > 0.05). The correlation coefficients between fecal egg counts (FEC) x mast cells (r = -0.490; P < 0.05) and FEC x eosinophils in sub-mucosa (r = -0.714; P < 0.01) was significant in the Santa Ines sheep. In the Ile de France group, the correlation coefficients between globule leucocytes x FEC (r = -0.879; P < 0.001) and histamine x worm burden (r = -0.833; P < 0.01) were also significant. In the Santa Ines and Ile de France sheep, correlation coefficients between IgA anti-L3 x worm burden and IgA anti-L3 x FEC were negative. In general, inflammatory cells and IgA-parasite-specific in abomasum were inversely associated with H. contortus worm burden and FEC indicating that they may impair parasite development or fecundity in the three breeds of sheep. However, similar mean values of inflammatory cells and IgA were found in the resistant (Santa Ines) and in the susceptible (Suffolk and Ile de France) breeds of sheep. The enumeration of cells by histological assessment does not provide information on their functional activity, which may be different among breeds. Thus, the effect of breed on the functional activity of these and other inflammatory cells is an important area for further study. (c) 2004 Elsevier B.V. All rights reserved.

Influence of dietary protein supply on resistance to experimental infections with Haemonchus contortus in Ile de France and Santa Ines lambs

The effect of Haemonchus contortus infection in sheep fed with a moderate and high protein content diet was evaluated in two breeds of sheep. Forty-eight Ile de France and Santa Ines lambs were maintained indoors since birth, in worm-free conditions. The lambs were allocated after weaning in four groups of six animals per breed, which were either infected or remain uninfected and given access to either a moderately or highly metabolizable protein diet. The moderately and highly metabolizable protein diets were calculated to supply 75 and 129 g metabolizable protein per kg of dry matter (MP/kg DM), respectively. The infection consisted of a trickle infection with 300 infective larvae, three times a week, for 12 weeks. Significant differences were observed for mast cell, globule leukocyte and eosinophil counts in the abomasal mucosa of the infected groups compared to the control of both breeds (P < 0.05), regardless of the diet supplied. Significantly higher IgA anti-L5 antibody was detected in the infected Santa Ines groups than in the infected Ile de France groups (P < 0.05). Increased metabolizable protein supply resulted in larger body weight gain and higher packed cell volumes for both breeds (P < 0.05). Both breeds showed an increased ability to withstand the pathophysiological effects of H. contortus infection when given access to the highly metabolizable protein diet. However, increased metabolizable protein supply resulted in reduced worm burdens in Santa Ines lambs but not in the Ile de France lambs (P < 0.05). The present results show that the increase in protein content in growing lamb diets may benefit resistance and resilience to gastrointestinal parasites but that these benefits may vary among breeds. (c) 2005 Elsevier B.V. All rights reserved.

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-1 2p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC- 1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (1-3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P < 0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P < 0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-1 2p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by TH2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group. (C) 2008 Elsevier B.V. All rights reserved.

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