title: MUTZ-3-derived dendritic cells as an in vitro alternative model to CD34+ progenitor-derived dendritic cells for testing of chemical sensitizers.
abstract: The cytokine-dependent CD34(+) human acute myeloid leukaemia cell line MUTZ-3 was used to generate immature dendritic-like cells (MUTZ-3 DC) and their validity as an alternative to primary CD34(+) progenitor-derived DC (CD34-DC) for testing chemical-induced sensitization was assessed. Expression levels of the DC maturation markers HLA-DR, CD86, CD83 and CD11c were studied using flow cytometry after 24 and 48 h exposure to the model compound nickel sulphate (100 and 300 microM). No maturation of MUTZ-3 DC was observed, whereas significantly upregulated expression levels of CD83 and CD86 were noticed in CD34-DC after 24h treatment with 300 microM nickel sulphate compared to control cells. Differential expression of the cytokine genes IL1beta, IL6, IL8, CCL2, CCL3, CCL3L1, CCL4 was analyzed using real-time RT-PCR after 6, 10 and 24h of nickel sulphate exposure. In response to 100 microM nickel sulphate MUTZ-3 DC revealed slightly upregulated mRNA levels after 24h, whereas 300 microM induced transcription of CCL3, CCL3L1 and IL8 significantly after 6 or 10h. These cytokine data correspond to the previously observed effects of 100 microM nickel sulphate in CD34-DC. Our findings underline the stimulatory capacity of nickel sulphate in MUTZ-3 DC with regard to cytokine mRNA induction, but not surface marker expression. Compared to CD34-DC, however, the studied endpoint markers seemed to be less inducible, making the MUTZ-3 DC model in its presented form less suitable for in vitro testing of sensitization. Further assessment of MUTZ-3 DC using other differentiation protocols and an extended set of chemicals will be required to reveal whether this cell line may be a valid alternative model system to primary CD34-DC.
authors.affiliation: Flemish Institute for Technological Research (VITO NV), Environmental Risk and Health, BE-2400 Mol, Belgium. inge.nelissen@vito.be
authors.forename: Inge
authors.initials: I
authors.lastname: Nelissen
authors.forename: Ingrid
authors.initials: I
authors.lastname: Selderslaghs
authors.forename: Rosette Van Den
authors.initials: RV
authors.lastname: Heuvel
authors.forename: Hilda
authors.initials: H
authors.lastname: Witters
authors.forename: Geert R
authors.initials: GR
authors.lastname: Verheyen
authors.forename: Greet
authors.initials: G
authors.lastname: Schoeters
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D045744:Cell Line, Tumor; D016207:Cytokines / Q000235:genetics; D003713:Dendritic Cells / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D009532:Nickel / Q000633:toxicity*; D010641:Phenotype
chemical_list: D018952:Antigens, CD34; D016207:Cytokines; C029938:nickel sulfate; D009532:Nickel