Differential Protein Pathways in 1,25-Dihydroxyvitamin D-3 and Dexamethasone Modulated Tolerogenic Human Dendritic Cells

Gabriela Bomfim Ferreira, Fleur S. Kleijwegt, Etienne Waelkens, Kasper Lage Hansen, Tatjana Nikolic, Daniel Aaen Hansen, Christopher Workman, Bart O. Roep, Lut Overbergh, Chantal Mathieu

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    Abstract

    Tolerogenic dendritic cells (DC) that are maturation-resistant and locked in a semimature state are promising tools in clinical applications for tolerance induction. Different immunomodulatory agents have been shown to induce a tolerogenic DC phenotype, such as the biologically active form of vitamin D (1,25(OH)(2)D-3), glucocorticoids, and a synergistic combination of both. In this study, we aimed to characterize the protein profile, function and phenotype of DCs obtained in vitro in the presence of 1,25(OH)(2)D-3, dexamethasone (DEX), and a combination of both compounds (combi). Human CD14(+) monocytes were differentiated toward mature DCs, in the presence or absence of 1,25(OH)(2)D-3 and/or DEX. Cells were prefractionated into cytoplasmic and microsomal fractions and protein samples were separated in two different pH ranges (pH 3-7NL and 6-9), analyzed by 2D-DIGE and differentially expressed spots (p <0.05) were identified after MALDI-TOF/TOF analysis. In parallel, morphological and phenotypical analyses were performed, revealing that 1,25(OH)(2)D-3- and combi-mDCs are closer related to each other than DEX-mDCs. This was translated in their protein profile, indicating that 1,25(OH)(2)D-3 is more potent than DEX in inducing a tolerogenic profile on human DCs. Moreover, we demonstrate that combining 1,25(OH)(2)D-3 with DEX induces a unique protein expression pattern with major imprinting of the 1,25(OH)(2)D-3 effect. Finally, protein interaction networks and pathway analysis suggest that 1,25(OH)(2)D-3, rather than DEX treatment, has a severe impact on metabolic pathways involving lipids, glucose, and oxidative phosphorylation, which may affect the production of or the response to ROS generation. These findings provide new insights on the molecular basis of DC tolerogenicity induced by 1,25(OH)(2)D-3 and/or DEX, which may lead to the discovery of new pathways involved in DC immunomodulation.
    Original languageEnglish
    JournalJournal of Proteome Research
    Volume11
    Issue number2
    Pages (from-to)941-971
    ISSN1535-3893
    DOIs
    Publication statusPublished - 2012

    Keywords

    • clinical application
    • differential protein pathway
    • immune response
    • metabolic pathway
    • molecular basis
    • oxidative phosphorylation
    • protein interaction network
    • protein profile
    • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
    • 1,25-dihydroxyvitamin D3
    • dexannethasone DEX
    • glucocorticoids
    • glucose 58367-01-4
    • lipids
    • vitamin D 1406-16-2
    • 02506, Cytology - Animal
    • 02508, Cytology - Human
    • 10060, Biochemistry studies - General
    • 10063, Biochemistry studies - Vitamins
    • 10066, Biochemistry studies - Lipids
    • 10067, Biochemistry studies - Sterols and steroids
    • 10068, Biochemistry studies - Carbohydrates
    • 15002, Blood - Blood and lymph studies
    • 15004, Blood - Blood cell studies
    • 34502, Immunology - General and methods
    • Chemical Coordination and Homeostasis
    • CD14 cell immune system
    • dendritic cell immune system
    • monocyte immune system, blood and lymphatics
    • peripheral blood mononuclear cell immune system, blood and lymphatics
    • 2D-DIGE electrophoretic techniques, laboratory techniques
    • MALDI-TOF/TOF analysis laboratory techniques, spectrum analysis techniques
    • Biochemistry and Molecular Biophysics
    • Immune System

    Cite this

    Ferreira, G. B., Kleijwegt, F. S., Waelkens, E., Hansen, K. L., Nikolic, T., Hansen, D. A., Workman, C., Roep, B. O., Overbergh, L., & Mathieu, C. (2012). Differential Protein Pathways in 1,25-Dihydroxyvitamin D-3 and Dexamethasone Modulated Tolerogenic Human Dendritic Cells. Journal of Proteome Research, 11(2), 941-971. https://doi.org/10.1021/pr200724e