title: Optimization of the IL-8 Luc assay as an in vitro test for skin sensitization.
abstract: We previously reported a dataset of the IL-8 Luc assay covering reference chemicals published by ECVAM, in which the effects of chemicals on IL-8 promoter activity were evaluated by an IL-8 reporter cell line, THP-G8 cells. To clarify its performance, we created another dataset of 88 sensitizers and 34 non-sensitizers. Simultaneously, to improve its performance, we changed the incubation time from 5 h to 16 h, deleted the criterion regarding the effects of N-acetylcysteine, and set an exclusion criterion for detergents. These modifications significantly improved its performance. In addition, we examined the following three criteria to judge chemicals as sensitizers: Criterion 1: Fold induction of SLO luciferase activity (FlnSLO-LA)⩾1.4, Criterion 2: the lower limit of the 95% confidence interval of FInSLO-LA⩾1.0, Criterion 3: the intersection of criteria 1 and 2. Among them, Criterion 1 produced the best performance, demonstrating that the accuracy, sensitivity and specificity were 81%, 79%, and 90%, respectively. In addition, we found that the IL-8 Luc assay solubilizing chemicals with X-VIVO substantially improved its performance. Finally, the IL-8 Luc assay combined with DPRA and DEREK could improve substantially its performance. These data suggest that the IL-8 Luc assay is a promising test method to screen skin sensitizers.
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kimura
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Chizu
authors.initials: C
authors.lastname: Fujimura
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Toshiya
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Nakajima
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmiya
authors.affiliation: Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan; Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa 761-0395, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan. Electronic address: saiba@med.tohoku.ac.jp.
authors.forename: Setsuya
authors.initials: S
authors.lastname: Aiba
mesh_terms: D000485:Allergens / Q000633:toxicity*; D045744:Cell Line, Tumor; D006241:Haptens / Q000633:toxicity*; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D012882:Skin Tests
chemical_list: D000485:Allergens; D006241:Haptens; D016209:Interleukin-8; D008156:Luciferases
keywords: Alternative method; Contact dermatitis; Luciferase assay; Sensitization test; Skin sensitization