Proteome analysis demonstrates profound alterations in human dendritic cell nature by TX527, an analogue of vitamin D |
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Authors: | Gabriela Bomfim Ferreira Evelyne van Etten Kasper Lage Daniel Aaen Hansen Yves Moreau Christopher T Workman Mark Waer Annemieke Verstuyf Etienne Waelkens Lut Overbergh Chantal Mathieu |
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Institution: | 1. Laboratory for Experimental Medicine and Endocrinology (LEGENDO), University Hospital Gasthuisberg, Catholic University of Leuven, Leuven, Belgium;2. Center for Biological Sequence Analysis, Technical University of Denmark, Lyngby, Denmark;3. Pediatric Surgical Research Laboratories, MassGeneral Hospital for Children, Massachusetts General Hospital, Boston, MA, USA;4. Harvard Medical School, Boston, MA, USA;5. ESAT–SDC, Department of Electrical Engineering, Catholic University of Leuven, Heverlee, Belgium;6. Experimental Transplantation Section, University Hospital Gasthuisberg, Catholic University of Leuven, Leuven, Belgium;7. ProMeta, University Hospital Gasthuisberg, Catholic University of Leuven, Leuven, Belgium;8. Laboratory of Protein Phosphorylation and Proteomics, University Hospital Gasthuisberg, Catholic University of Leuven, Leuven, Belgium |
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Abstract: | Structural analogues of vitamin D have been put forward as therapeutic agents able to exploit the immunomodulatory effects of vitamin D, without its undesired calcemic side effects. We have demonstrated that TX527 affects dendritic cell (DC) maturation in vitro, resulting in the generation of a tolerogenic cell. In the present study, we aimed to explore the global protein changes induced by the analogue in immature DC (iDC) and mature human DC and to correlate them with alterations in DC morphology and function. Human CD14+ monocytes were differentiated toward iDC or mature DCs, in the presence or absence of TX527 (10?8 M) (n=4). Protein samples were separated into two different pH ranges (pH4–7 and 6–9), analyzed by 2‐D DIGE and differentially expressed spots (p<0.01) were identified by MALDI‐TOF/TOF (76.3 and 70.7% in iDC and mature DCs, respectively). Differential protein expression revealed three protein groups predominantly affected by TX527 treatment, namely proteins involved in cytoskeleton structure, in protein biosynthesis/proteolysis and in metabolism. Moreover, protein interactome‐network analysis demonstrated close interaction between these different groups (p<0.001) and morphological and functional analyses confirmed the integrated effect of TX527 on human DCs, resulting in a cell with altered morphology, cell surface marker expression, endocytic and migratory capacity. |
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Keywords: | 2‐D DIGE Dendritic cell Immunotolerance Network Vitamin D analogue |
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