title: Proteome analysis demonstrates profound alterations in human dendritic cell nature by TX527, an analogue of vitamin D.
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.
authors.affiliation: Laboratory for Experimental Medicine and Endocrinology (LEGENDO), University Hospital Gasthuisberg, Catholic University of Leuven, B-3000 Leuven, Belgium.
authors.forename: Gabriela Bomfim
authors.initials: GB
authors.lastname: Ferreira
authors.forename: Evelyne
authors.initials: E
authors.lastname: van Etten
authors.forename: Kasper
authors.initials: K
authors.lastname: Lage
authors.forename: Daniel Aaen
authors.initials: DA
authors.lastname: Hansen
authors.forename: Yves
authors.initials: Y
authors.lastname: Moreau
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Workman
authors.forename: Mark
authors.initials: M
authors.lastname: Waer
authors.forename: Annemieke
authors.initials: A
authors.lastname: Verstuyf
authors.forename: Etienne
authors.initials: E
authors.lastname: Waelkens
authors.forename: Lut
authors.initials: L
authors.lastname: Overbergh
authors.forename: Chantal
authors.initials: C
authors.lastname: Mathieu
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D020543:Proteome / Q000032:analysis*; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014815:Vitamins / Q000494:pharmacology*
chemical_list: D020543:Proteome; D014815:Vitamins; D014807:Vitamin D