The cytokine-dependent MUTZ-3 cell line as an in vitro model for the screening of contact sensitizers.
Langerhans cells (LC) are key mediators of contact allergenicity in the skin. However, no in vitro methods exist which are based on the activation process of LC to predict the sensitization potential of chemicals. In this study, we have evaluated the performances of MUTZ-3, a cytokine-dependent human monocytic cell line, in its response to sensitizers. First, we compared undifferentiated MUTZ-3 cells with several standard human cells such as THP-1, KG-1, HL-60, K-562, and U-937 in their response to the strong sensitizer DNCB and the irritant SDS by monitoring the expression levels of HLA-DR, CD54, and CD86 by flow cytometry. Only MUTZ-3 and THP-1 cells show a strong and specific response to sensitizer, while other cell lines showed very variable responses. Then, we tested MUTZ-3 cells against a wider panel of sensitizers and irritants on a broader spectrum of cell surface markers (HLA-DR, CD40, CD54, CD80, CD86, B7-H1, B7-H2, B7-DC). Of these markers, CD86 proved to be the most reliable since it detected all sensitizers, including benzocaine, a classical false negative in local lymph node assay (LLNA) but not irritants. We confirmed the MUTZ-3 response to DNCB by real-time PCR analysis. Taken together, our data suggest that undifferentiated MUTZ-3 cells may represent a valuable in vitro model for the screening of potential sensitizers.
Azam
Philippe
P
AFSSAPS, Unité BCM, 635 Rue de la Garenne, 34740 Vendargues, France.
Peiffer
Jean-Luc
JL
Chamousset
Delphine
D
Tissier
Marie-Hélène
MH
Bonnet
Pierre-Antoine
PA
Vian
Laurence
L
Fabre
Isabelle
I
Ourlin
Jean-Claude
JC
D000328:Adult; D000954:Antigens, Surface / Q000096:biosynthesis; D018122:B7-1 Antigen / Q000494:pharmacology; D051940:B7-2 Antigen / Q000235:genetics; D019013:CD40 Antigens / Q000096:biosynthesis; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D003470:Culture Media; D016207:Cytokines / Q000502:physiology*; D003877:Dermatitis, Contact / Q000503:physiopathology*; D004137:Dinitrochlorobenzene / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000235:genetics; D006801:Humans; D060945:Inducible T-Cell Co-Stimulator Ligand; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D007509:Irritants / Q000494:pharmacology / Q000633:toxicity*; D008297:Male; D011237:Predictive Value of Tests; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
D000954:Antigens, Surface; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D019013:CD40 Antigens; D003470:Culture Media; D016207:Cytokines; D004137:Dinitrochlorobenzene; D006684:HLA-DR Antigens; D060945:Inducible T-Cell Co-Stimulator Ligand; D007509:Irritants; D018799:Intercellular Adhesion Molecule-1; D012967:Sodium Dodecyl Sulfate; D012313:RNA