title: Differential modulation of CXCR4 and CD40 protein levels by skin sensitizers and irritants in the FSDC cell line.
abstract: The development of non-animal methods for skin sensitization testing is an urgent challenge. Some of the most promising in vitro approaches are based on the analysis of phenotypical and functional modifications induced by sensitizers in dendritic cell models. In this work, we evaluated, for the first time, a fetal skin-derived dendritic cell line (FSDC) as a model to discriminate between sensitizers and irritants, through analysis of their effects on CD40 and CXCR4 protein expression. The chemicals concentrations were chosen based on a slight cytotoxicity effect (up to 15%). Protein levels were evaluated by Western blot and immunocytochemistry, after stimulation with the skin sensitizers 2,4-dinitrofluorobenzene (DNFB), 1,4-phenylenediamine (PPD) and nickel sulphate (NiSO(4)), the non-sensitizer 2,4-dichloronitrobenzene (DCNB), and the irritants sodium dodecyl sulphate (SDS) and benzalkonium chloride (BC). All sensitizers tested increased CD40 and CXCR4 levels. In contrast, irritants decreased both proteins levels, with a more pronounced effect on CXCR4. In agreement with these results, dendritic cells derived from human peripheral blood monocytes-derived dendritic cells (MoDC) showed a similar response pattern to the skin sensitizer and irritant tested, PPD and SDS, respectively. In conclusion, evaluation of CD40 and CXCR4 proteins in chemical-treated FSDC may represent a useful tool in a future in vitro test for sensitizing assessment.
authors.affiliation: Faculdade de Farmácia, Universidade de Coimbra, Rua do Norte, Coimbra, Portugal.
authors.forename: Bruno Miguel
authors.initials: BM
authors.lastname: Neves
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Cruz
authors.forename: Vera
authors.initials: V
authors.lastname: Francisco
authors.forename: Margarida
authors.initials: M
authors.lastname: Gonçalo
authors.forename: Américo
authors.initials: A
authors.lastname: Figueiredo
authors.forename: Carlos B
authors.initials: CB
authors.lastname: Duarte
authors.forename: Maria Celeste
authors.initials: MC
authors.lastname: Lopes
mesh_terms: D000485:Allergens / Q000145:classification / Q000633:toxicity*; D000826:Animal Testing Alternatives; D000818:Animals; D001548:Benzalkonium Compounds / Q000633:toxicity; D019013:CD40 Antigens / Q000378:metabolism*; D002460:Cell Line; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004139:Dinitrofluorobenzene / Q000633:toxicity; D006801:Humans; D007509:Irritants / Q000145:classification / Q000633:toxicity*; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D051379:Mice; D009532:Nickel / Q000633:toxicity; D009578:Nitrobenzenes / Q000633:toxicity; D010655:Phenylenediamines / Q000633:toxicity; D019718:Receptors, CXCR4 / Q000378:metabolism*; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity
chemical_list: D000485:Allergens; D001548:Benzalkonium Compounds; D019013:CD40 Antigens; C514936:CXCR4 protein, mouse; D007509:Irritants; D009578:Nitrobenzenes; D010655:Phenylenediamines; D019718:Receptors, CXCR4; D012967:Sodium Dodecyl Sulfate; C029938:nickel sulfate; D009532:Nickel; C053747:2,4-dichloro-1-nitrobenzene; D004139:Dinitrofluorobenzene; C029728:4-phenylenediamine