title: Modulation of phenotype, cytokine production and stimulatory function of CD34+-derived DC by NiCl(2) and SDS.
abstract: One of the most promising alternatives to identify the sensitizing potency of new products is the in vitro culture of human dendritic cells (DC). In vivo, dendritic cells present in the skin are highly specialized antigen-presenting cells (APC) of the immune system, which play a crucial role in the induction of allergic reactions. The DC produce specific cytokines and upregulate specific co-stimulatory molecules in addition to the antigen-MHC complex in order to promote an optimal T-cell activation. The aim of our study is to assess the phenotype, cytokine production and autologous T-cell stimulatory capacity of the in vitro CD34+-derived dendritic cells after 24 hours of incubation with the metal allergen NiCl(2) (100-300 microM) and the irritant sodium dodecyl sulfate (SDS; 0.01%), in order to find a sensitive endpoint to discriminate between sensitizers and irritants. After exposure to Ni, a significant increase in the number of CD83+ and CD86+ cells was noticed. The intensity of CD86 as well as the intensity of the HLA-DR molecule on the DC also showed a significant increase. The expression of the co-stimulatory molecule CD80 was not changed after exposure. SDS was not able to increase the expression of any of the analysed markers. The production of IL-6 increased significantly after exposure of dendritic cells to Ni, but not after SDS exposure. Results on tumor necrosis factor-alpha (TNF-alpha) production are somewhat equivocal. Although not statistically significant, TNF-alpha was upregulated in one out of three experiments after 48 hours of exposure to the Ni allergen, but increases were also noticed after exposure to SDS (two out of three experiments). Both exposure to Ni and SDS caused an upregulation (not significant) in the IL-12 production by DC, but the production was higher in Ni-exposed DC compared to SDS-exposed cells. In none of the exposed DC cultures was it possible to detect IL-1 beta. The antigen-presenting capacity of the DC in autologous MLR could not be demonstrated. Nevertheless, T-cell proliferation after DC stimulation was noticed in allogenic MLR.
authors.affiliation: Vito, Flemish Institute for Technological Research, Boeretang 200, 2400 Mol, Belgium. ann.desmedt@vito.be
authors.forename: A C
authors.initials: AC
authors.lastname: De Smedt
authors.forename: R L
authors.initials: RL
authors.lastname: Van Den Heuvel
authors.forename: N
authors.initials: N
authors.lastname: Zwi Berneman
authors.forename: G E
authors.initials: GE
authors.lastname: Schoeters
mesh_terms: D000485:Allergens / Q000633:toxicity; D000826:Animal Testing Alternatives; D018952:Antigens, CD34 / Q000032:analysis; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis*; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005312:Fetal Blood; D005434:Flow Cytometry; D006801:Humans; D007231:Infant, Newborn; D007509:Irritants / Q000633:toxicity; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects*; D007959:Lymphocyte Culture Test, Mixed; D009532:Nickel / Q000633:toxicity*; D010641:Phenotype; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity*; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D000485:Allergens; D018952:Antigens, CD34; D016207:Cytokines; D007509:Irritants; D008070:Lipopolysaccharides; D012967:Sodium Dodecyl Sulfate; C022838:nickel chloride; D009532:Nickel