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Phenotypic and functional comparison of two distinct subsets of programmable cell of monocytic origin (PCMOs)-derived dendritic cells with conventional monocyte-derived dendritic cells

Cellular & Molecular Immunology volume 13, pages 160–169 (2016)Cite this article

Abstract

Dendritic cells (DCs) are professional antigen-presenting cells with the ability to induce primary T-cell responses. They are commonly produced by culturing monocytes in the presence of IL-4 and GM-CSF (cells produced in this manner are called conventional DCs). Here we report the generation of two functionally distinct subsets of DCs derived from programmable cells of monocytic origin (PCMOs) in the presence of IL-3 or tumor necrosis factor alpha (TNF-α). Monocytes were treated with macrophage colony-stimulating factor (M-CSF) and IL-3 for 6 days and then incubated with IL-4 and IL-3 (for IL-3 DCs) or with IL-4, GM-CSF and TNF-α (for TNF-α DCs) for 7 days. Monocytes were then loaded with tumor lysate (used as antigen), and poly (I∶C) was added. The maturation factors TNF-α and monocyte conditioned medium (MCM) were added on days 4 and 5, respectively. The phenotypes of the DCs generated were characterized by flow cytometry, and the cells' phagocytic activities were measured using FITC-conjugated latex bead uptake. T-cell proliferation and cytokine release were assayed using MTT and commercially available ELISA kits, respectively. We found that either IL-3DCs or TNF-α DCs induce T-cell proliferation and cytokine secretion; the cytokine release pattern showed reduced IL-12/IL-10 and IFN-γ/IL-4 ratios in both types of DCs and in DC-primed T-cell supernatant, respectively, which confirmed that the primed T cells were polarized toward aTh2-type immune response. We concluded that PCMOs are a new cell source that can develop into two functionally distinct DCs that both induce a Th2-type response in vitro. This modality can be used as a DC-based immunotherapy for autoimmune diseases.

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Acknowledgements

This work was supported by the Institute of Biotechnology of Urmia University (IBUU), Grant No. 8-D-92. We thank Professor Andreas Nüssler, Dr Mahmood Bozorgmehr and Dr Reza Habibian for technical advice; Mr Asghar Aliyari and Mr Yousef Heidar Sani (Immunology Laboratory, Veterinary School of Urmia University, Urmia, Iran) and Mr Ehsan Janzamin and Mr FazelSamani (Department of Stem Cell, Flow Cytometry Laboratory, Royan Institute, Tehran, Iran) for technical assistance; and Mr Yones Najafi Darmian and the other volunteers in this study.

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  1. Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

    Babak Beikzadeh

  2. Department of Cellular and Molecular Biotechnology, Institute of Biotechnology, Urmia University, Urmia, Iran

    Nowruz Delirezh

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  1. Babak Beikzadeh
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Correspondence to Nowruz Delirezh.

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Beikzadeh, B., Delirezh, N. Phenotypic and functional comparison of two distinct subsets of programmable cell of monocytic origin (PCMOs)-derived dendritic cells with conventional monocyte-derived dendritic cells. Cell Mol Immunol 13, 160–169 (2016). https://doi.org/10.1038/cmi.2014.135

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