Optimization of the IL-8 Luc assay as an in vitro test for skin sensitization

•We created another data set of 122 chemicals of the IL-8 Luc assay.•We analyzed the reason for poor performance of the original protocol.•The solubilizing chemicals with X-VIVO 15 significantly improved its performance.•Its performance was significantly improved, 90% of accuracy and 90% of specific...

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Bibliographic Details
Published in:Toxicology in vitro 2015-10, Vol.29 (7), p.1816-1830
Main Authors: Kimura, Yutaka, Fujimura, Chizu, Ito, Yumiko, Takahashi, Toshiya, Nakajima, Yoshihiro, Ohmiya, Yoshihiro, Aiba, Setsuya
Format: Article
Language:English
Subjects:
Allergens - toxicity
Alternative method
Cell Line, Tumor
Contact dermatitis
Haptens - toxicity
Humans
Interleukin-8 - genetics
Luciferase assay
Luciferases - genetics
Sensitization test
Skin sensitization
Skin Tests
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Summary:•We created another data set of 122 chemicals of the IL-8 Luc assay.•We analyzed the reason for poor performance of the original protocol.•The solubilizing chemicals with X-VIVO 15 significantly improved its performance.•Its performance was significantly improved, 90% of accuracy and 90% of specificity. 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 5h to 16h, 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.
ISSN:0887-2333
1879-3177
DOI:10.1016/j.tiv.2015.07.006