RESUMO CONTEXTO: Durante a infecção por Helicobacter pylori (HP), a infiltração dos leucócitos na mucosa estomacal é dirigida por quimiocinas produzidas localmente que desempenham um papel decisivo no resultado da infecção. O CagA é o fator de virulência mais potente do HP, de modo que a infecção com cepas CagA + está associada a complicações mais graves do que a infecção com CagA - HP. OBJETIVO: O objetivo foi determinar a expressão das quimiocinas CXCL10, CCL17, CCL20 e CCL22, e seus receptores por CagA + HP- e CagA - extrato bruto (EB) derivado de HP (HP-EB) de células mononucleares do sangue periférico (CMSP) de pacientes com úlcera péptica (UP). MÉTODOS: O soro e as CMSP foram coletados de 20 pacientes com UP infectados pelo HP, 20 indivíduos assintomáticos infectados pelo HP (AI-HP) e 20 indivíduos saudáveis não infectados pelo HP (NI-HP). As CMSP foram cultivadas na ausência de estimulador ou com extrato bruto CagA + HP de 10 μg (CagA + EB), 10 μg CagA - extrato bruto HP (CagA - EB). Quimiocinas e receptores foram medidos por ELISA e PCR em tempo real, respectivamente. RESULTADOS: Em pacientes com UP a produção de quimiocinas CXCL10, CCL17, CCL20 e CCL22, e a expressão dos receptores de quimiocina CXCR3, CCR4 e CCR6 por CagA + CMSP induzidos pelo EB foram significativamente maiores do que as culturas não estimuladas e CagA - EB estimulados. A produção de CXCL10 por CagA + EB estimulou as CMSP de sujeitos AI-HP em proporção significativamente maior do que as culturas iguais dos grupos UP e NI-HP. A produção de CCL17 e CCL20 por grupos não estimulados, CagA + EB estimulado, e CagA - EB estimulou CMSP de sujeitos com UP e foram significativamente superiores às culturas iguais dos grupos NI-HP e AI-HP. A produção de CCL22 por grupos não estimulados, CagA + EB estimulado e CagA - EB estimulado pelo grupo NI-HP foram significativamente maiores do que as culturas iguais dos grupos AI-HP e PU. O CagA + EB estimulou as CMSP dos sujeitos do AI-HP, expressando menores quantidades de CCR6 em comparação com as CMSP estimuladas pelo CagA + EB de grupos NI-HP e UP. Os níveis sanguíneos de CXCL10 e CCL20 nos grupos UP e AI-HP foram significativamente superiores aos dos sujeitos do NI-HP. Os grupos NI-HP e AI-HP apresentaram níveis sanguíneos mais elevados de CCL22 em comparação com pacientes com UP. CONCLUSÃO: Os resultados indicaram que o estado CagA da bactéria influencia a expressão de quimiocinas e receptores por HP-EB estimulados CMSP de pacientes com UP.

ABSTRACT BACKGROUND: During the Helicobacter pylori (HP) infection, the infiltration of the leukocytes into stomach mucosa is directed by locally produced chemokines that play a decisive role in infection outcome. The CagA is the most potent virulence factor of HP, so that the infection with CagA + strains is associated with more severe complications than infection with CagA - HP. OBJECTIVE: The aim was to determine the expression of chemokines CXCL10, CCL17, CCL20 and CCL22, and their receptors by CagA + HP- and CagA - HP-derived crude extract (HP-CE)-stimulated peripheral blood mononuclear cells (PBMCs) from peptic ulcer (PU) patients. METHODS: The serum and the PBMCs were collected from 20 HP-infected PU patients, 20 HP-infected asymptomatic subjects (HIA) and 20 non-infected healthy subjects (NHS). The PBMCs were cultured in absence of stimulator or with 10 µg CagA + HP crude extract (CagA + CE), 10 µg CagA - HP crude extract (CagA - CE). Chemokines and receptors were measured by ELISA and real time-PCR respectively. RESULTS: In PU patients, the production of chemokines CXCL10, CCL17, CCL20 and CCL22, and the expression of chemokine receptors CXCR3, CCR4 and CCR6 by CagA + CE-induced PBMCs were significantly higher than non-stimulated and CagA - CE stimulated cultures. The CXCL10 production by CagA + CE stimulated PBMCs from HIA subjects was significantly higher than the equal cultures from PU and NHS groups. The CCL17 and the CCL20 production by non-stimulated, CagA + CE stimulated, and CagA - CE stimulated PBMCs from PU subjects were significantly higher than the equal cultures from NHS and HIA groups. The CCL22 production by non-stimulated, CagA + CE stimulated and CagA - CE stimulated PBMCs from NHS group were significantly higher than the equal cultures from HIA and PU groups. The CagA + CE stimulated PBMCs from HIA subjects expressed lower amounts of CCR6 in comparison with CagA + CE stimulated PBMCs from NHS and PU groups. The serum levels CXCL10 and CCL20 in PU and HIA groups were significantly higher than NHS subjects. NHS and HIA groups displayed higher serum levels of CCL22 in comparison with PU patients. CONCLUSION: Results indicated that the CagA status of bacterium influence the expression of chemokines and receptors by HP-CE stimulated PBMCs from PU patients.

It has been reported that the immune system undergoes age and gender changes. The aim of this study was to investigate the age- and gender-dependent changes of salivary IgA and IgE levels among healthy subjects. A total of 203 healthy individuals (aged 1-70 years) were enrolled in the study. Two milliliters of saliva were collected from all participants, and salivary IgA and IgE levels were measured by the ELISA technique. Mean salivary IgA levels were significantly higher in subjects aged 11-20 years as compared to subjects aged 1-10 years (P < 0.01). Mean salivary IgA levels increased with age up to the age of 60 years, and then slightly decreased in subjects aged 61-70 years. The frequency of subjects with detectable levels of salivary IgE and mean salivary IgE levels gradually increased with age, with maximum levels being observed in the 31-40 years age group and not changing significantly thereafter. The mean levels of salivary IgA and IgE in adults were significantly higher than those observed in children (P < 0.00001 and P < 0.05, respectively). No significant differences were observed between men and women regarding both salivary immunoglobulins. These results showed age-dependent changes of the salivary IgA and IgE levels. Gender had no effect on the salivary levels of IgA and IgE.