title: Evaluation of the skin sensitization potential of chemicals in THP-1/keratinocyte co-cultures.
abstract: Many attempts have been made to develop in vitro sensitization tests that employ dendritic cells (DCs), DC-like cell lines or keratinocytes. The aim of the present investigation was to establish a co-culture of THP-1 cells and keratinocytes for evaluation of skin sensitization potential of chemicals. Co-cultures were constructed by THP-1 cells cultured in lower compartments and keratinocytes cultured in upper compartments of cell culture inserts. After 24 h exposure to sensitizers (2, 4-dinitrochlorobenzene, p-phenylenediamine, formaldehyde, nickel sulfate, isoeugenol and eugenol) and non-sensitizers (sodium lauryl sulfate, benzalkonium chloride and lactic acid), the expression of CD86 and CD54 on THP-1 cells were evaluated by flow cytometry, and cell viabilities were determined. The sensitizers induced the augmentation of CD86 and CD54 expression, but the non-sensitizers had no significant effect. Compared with mono-cultures of THP-1 cells, the augmentation of CD86 and CD54 could be detected even at a non-toxic concentration of sensitizers in THP-1 cell/keratinocyte co-cultures. Moreover, isoeugenol was distinguished as a sensitizer in co-cultures, but failed to be identified in mono-cultures. These results revealed that the co-cultures of THP-1 cells and keratinocytes were successfully established and suitable for identifying sensitizers using CD86 and CD54 expression as identification markers.
authors: Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, Nanjing, China.
authors: Yu-Ping
authors: YP
authors: Cao
authors: Peng-Cheng
authors: PC
authors: Ma
authors: Wei-Da
authors: WD
authors: Liu
authors: Wu-Qing
authors: WQ
authors: Zhou
authors: Yue
authors: Y
authors: Tao
authors: Meng-Li
authors: ML
authors: Zhang
authors: Ling-Jun
authors: LJ
authors: Li
authors: Zi-Yi
authors: ZY
authors: Chen
mesh_terms: D000826:Animal Testing Alternatives / Q000379:methods*; D051940:B7-2 Antigen / Q000378:metabolism; D001548:Benzalkonium Compounds / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D017449:Dermatitis, Allergic Contact / Q000276:immunology*; D004137:Dinitrochlorobenzene / Q000276:immunology / Q000494:pharmacology; D005054:Eugenol / Q000031:analogs & derivatives / Q000276:immunology / Q000494:pharmacology; D005557:Formaldehyde / Q000276:immunology / Q000494:pharmacology; D006241:Haptens / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000276:immunology*; D019344:Lactic Acid / Q000276:immunology / Q000494:pharmacology; D009000:Monocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D009532:Nickel / Q000276:immunology / Q000494:pharmacology; D010655:Phenylenediamines / Q000276:immunology / Q000494:pharmacology; D012680:Sensitivity and Specificity; D012882:Skin Tests / Q000379:methods; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
chemical_list: D051940:B7-2 Antigen; D001548:Benzalkonium Compounds; C497964:CD86 protein, human; D004137:Dinitrochlorobenzene; D006241:Haptens; D010655:Phenylenediamines; D018799:Intercellular Adhesion Molecule-1; D005557:Formaldehyde; D019344:Lactic Acid; D012967:Sodium Dodecyl Sulfate; D005054:Eugenol; C029938:nickel sulfate; C036643:isoeugenol; D009532:Nickel; C029728:4-phenylenediamine