مقایسه تأثیر Tumor Necrosis Factor-α، Monocyte Conditioned Medium و polyinosinic-polycytidylic acid در بلوغ سلول‌های دندریتیک
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Abstract:
Abstract Background and purpose: Different laboratories have considered different maturation factors for producing human dendritic cells (DC) from peripheral blood monocytes. In this study, we comprehensively investigated the effect of adding poly (I:C) to standard maturation stimulus, i.e. MCM and TNF-α on the induction of T cell response. Materials and methods: Peripheral blood monocytes adhered to culture flask after two-hour incubation in the temperature 37 C° were cultured for four days with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Allogenic tumor antigen was added in the fourth day and in the fifth day, maturation factors including MCM and TNF-α were added to one group and MCM, TNF-α, and poly (I:C) to the other group. Morphology of the generated DCs was assessed using invert microscope and phenotypic analysis was carried out using anti CD14, HLA-DR and CD83 monoclonal antibodies. Furthermore, their functions were evaluated using induction of mixed lymphocyte reaction (MLR), phagocytic activity, and cytokine release by DC stimulated T lymphocytes. Results: It was found that dendritic cells with the same morphology were generated in the presence of MCM and TNF-α with or without poly (I:C). Flow cytometric analysis using anti-CD14, HLA-DR and CD83 revealed that addition of poly (I:C) to conventional maturation factors results in a decrease in the expression of CD14 and an increase in the expression of HLA-DR and CD83. Moreover, the amount of MLR slightly increased and the percentage of phagocytic cells decreased, while an increase was observed in their mean fluorescent intensity (MFI). Furthermore, the results revealed that adding poly (I:C) leads to the reduction of IL-12: IL-10 and IFN-γ: IL-4 ratios in DC and DC-primed T cell supernatants, respectively. Conclusion: Our results support this idea that adding poly (I:C) to MCM and TNF-α causes more maturation in dendritic cells and leads the stimulated lymphocytes to TH2 type cytokine release.
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Journal title
volume 21 issue 1
pages 49- 64
publication date 2012-03
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