title: The relationship between CD86/CD54 expression and THP-1 cell viability in an in vitro skin sensitization test--human cell line activation test (h-CLAT).
abstract: Recent regulations for cosmetics in Europe prohibit animal testing for evaluating the sensitization potential of chemicals to improve animal welfare. Yet, there is not an acceptable Organization for Economic Co-operation and Development non-animal skin sensitization test method. Several in vitro skin sensitization methods that focus on the activation of Langerhans cells, including human cell lines, are being evaluated as possible alternatives. In our previous study, we optimized our human cell line activation test (h-CLAT) using THP-1 cells (monocytic leukemia cell line) and conducted an inter-laboratory study. We found that measuring CD86/CD54 expression may be useful for predicting skin sensitization. The aim of this study was to confirm the relationship between CD86/CD54 expression and THP-1 cell viability in the h-CLAT. In this study, 21 allergens (e.g., dinitrochlorobenzene, p-phenylenediamine, Ni) and 8 non-allergens (e.g., SLS, lactic acid) were evaluated. For each chemical, more than 10 concentrations that gave a predicted cell viability range of 20-95% were used. The data showed that expression patterns of CD86/CD54 differed depending on chemical. For most allergens, cytotoxicity (65-90% cell viability) was needed for enhancement of CD86/CD54 expression. The criteria of "CD86 > or = 150 or CD54 > or = 200" resulted in an accuracy of 93%, which confirms appropriate cut-off criteria for h-CLAT. Furthermore, a good correlation was observed between EC3 of local lymph node assay and EC150(CD86) or EC200(CD54) of h-CLAT (12 or 16 chemicals, respectively), which would provide a useful estimate of allergic potency. These findings suggest that h-CLAT would be a good robust in vitro skin sensitization test.
authors.affiliation: Safety Science Research Laboratories, Kao Corporation, 2606 Akabane, Ichikai-Machi, Haga-Gun, Tochigi, Japan. sakaguchi.hitoshi@kao.co.jp
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sakaguchi
authors.forename: Takao
authors.initials: T
authors.lastname: Ashikaga
authors.forename: Masaaki
authors.initials: M
authors.lastname: Miyazawa
authors.forename: Nanae
authors.initials: N
authors.lastname: Kosaka
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Ito
authors.forename: Katsurako
authors.initials: K
authors.lastname: Yoneyama
authors.forename: Sakiko
authors.initials: S
authors.lastname: Sono
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Itagaki
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Toyoda
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Suzuki
mesh_terms: D000485:Allergens / Q000276:immunology / Q000494:pharmacology*; D000826:Animal Testing Alternatives / Q000379:methods; D000911:Antibodies, Monoclonal / Q000276:immunology; D051940:B7-2 Antigen / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000276:immunology; D003877:Dermatitis, Contact / Q000276:immunology / Q000473:pathology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology / Q000473:pathology*; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D011237:Predictive Value of Tests; D023422:Skin Irritancy Tests / Q000379:methods*; D012995:Solubility
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D051940:B7-2 Antigen; D018799:Intercellular Adhesion Molecule-1