Evaluation of two different dendritic cell preparations to BCG reactivity

Marek Fol, Anna Nitecka-Blaźlak, Piotr Szpakowski, Murty V.V.S. Madiraju, Wiesława Rudnicka, Magdalena Druszczyńska, Joël Pestel, Magdalena Kowalewicz-Kulbat

Abstract


Dendritic cells (DCs) play a key-role in the immune response against intracellular bacterial pathogens, including mycobacteria. Monocyte-derived dendritic cells (MoDCs) are considered to behave as inflammatory cell populations. Different immunomagnetic methods (positive and negative) can be used to purify monocytes before their in vitro differentiation and their culture behavior can be expected to be different. In this study we evaluated the reactivity of two dendritic cell populations towards the Bacillus Calmette–Guérin (BCG) antigen. Monocytes were obtained from the blood of healthy donors, using positive and negative immunomagnetic separation methods. The expression of DC-SIGN, CD86, CD80, HLA-DR and CD40 on MoDCs was estimated by flow cytometry. The level of IL-12p70, IL-10 and TNF-α was measured by ELISA. Neither of the tested methods affected the surface marker expression of DCs. No significant alteration in immunological response, measured by cytokine production, was noted either. After BCG stimulation, the absence of IL-12, but the IL-23 production was observed in both cell preparations. Positive and negative magnetic separation methods are effective techniques to optimize the preparation of monocytes as the source of MoDCs for potential clinical application

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