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Helicobacter ; 19(2): 136-43, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24495278

RESUMEN

BACKGROUND: Outer inflammatory protein A (OipA) has an important role in Helicobacter pylori pathogenesis. In this study, we purified the outer membrane protein and evaluated the effects of this protein on maturation and cytokine production by dendritic cells (DCs). MATERIALS AND METHODS: The oipA gene was inserted into pET28a, and this construct was transformed into Escherichia coli BL21 (DE3). Purification of the recombinant protein was performed by Ni-NTA affinity chromatography. Immature DCs were purified from spleen of C57BL/6 mice with more than 90% purity and were treated with several concentrations of OipA (1-20 µg/mL) overnight. Expression of maturation markers (CD86, CD40, and MHC-II) on the surface of DCs and production of IL-10 and IL-12 were assessed by flow cytometry and ELISA, respectively. RESULTS: The expression of DC maturation markers CD40, CD86, and MHC-II was downregulated on the surface of OipA-treated DCs at concentrations of 10 and 20 µg/mL compared with negative control. Production of IL-10 decreases with increasing OipA concentration at a concentration of 5 µg/mL, but we detected no change in IL-12 production. CONCLUSION: Inability to eliminate H. pylori from stomach is partly due to the evasion of the bacteria from the immune response. DCs are central mediators between innate and adaptive immunity, and DC cytokines direct the types of adaptive immune response. This study indicated that OipA of H. pylori is a DC maturation suppression factor. Previous studies have shown that H. pylori manage tolerogenic programming in DCs leading to long-time gastric colonization. In conclusion, H. pylori OipA helps the establishment of chronic infection with reduction in IL-10 and suppression of DC maturation.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/inmunología , Células Dendríticas/inmunología , Helicobacter pylori/inmunología , Evasión Inmune/inmunología , Proteínas Recombinantes/farmacología , Animales , Antígeno B7-2/biosíntesis , Proteínas de la Membrana Bacteriana Externa/genética , Antígenos CD40/biosíntesis , Células Cultivadas , Regulación hacia Abajo , Femenino , Expresión Génica/efectos de los fármacos , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/patología , Helicobacter pylori/patogenicidad , Antígenos de Histocompatibilidad Clase II/biosíntesis , Interleucina-10/biosíntesis , Interleucina-12/biosíntesis , Ratones , Ratones Endogámicos C57BL , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética
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