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In vitro suppression of dendritic cells by Helicobacter pylori OipA
Omid Teymournejad1, Ashraf M Mobarez, Zuhair M Hassan
1Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
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.
Insights
Helicobacter pylori outer inflammatory protein A (OipA) suppresses dendritic cell (DC) maturation and reduces IL-10 production. This immune evasion mechanism aids chronic H. pylori infection establishment.
Area of Science:
- Immunology
- Microbiology
- Cell Biology
Background:
- Outer inflammatory protein A (OipA) is crucial for Helicobacter pylori pathogenesis.
- Dendritic cells (DCs) bridge innate and adaptive immunity, directing immune responses.
Purpose of the Study:
- To purify OipA and investigate its impact on dendritic cell maturation and cytokine production.
- To understand OipA's role in H. pylori immune evasion.
Main Methods:
- Recombinant OipA was purified from E. coli using Ni-NTA affinity chromatography.
- Murine dendritic cells were treated with varying OipA concentrations (1-20 μg/mL).
- DC maturation markers (CD40, CD86, MHC-II) and cytokines (IL-10, IL-12) were analyzed via flow cytometry and ELISA.
Main Results:
- OipA downregulated DC maturation markers CD40, CD86, and MHC-II at 10 and 20 μg/mL.
- IL-10 production decreased with increasing OipA concentration (starting at 5 μg/mL).
- No significant change in IL-12 production was observed.
Conclusions:
- H. pylori OipA acts as a dendritic cell maturation suppression factor.
- OipA contributes to H. pylori's immune evasion and chronic infection establishment.
- Suppression of DC maturation and IL-10 reduction by OipA facilitates gastric colonization.
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