An international validation study of the IL-2 Luc assay for evaluating the potential immunotoxic effects of chemicals on T cells and a proposal for reference data for immunotoxic chemicals

Yutaka Kimura, Rie Yasuno, Mika Watanabe, Miwako Kobayashi, Tomoko Iwaki, Chizu Fujimura, Yoshihiro Ohmiya, Kohji Yamakage, Yoshihiro Nakajima, Mayumi Kobayashi, Nana Mashimo, Yumi Takagi, Takashi Omori, Emanuela Corsini, Dori Germolec, Tomoaki Inoue, Erwin L. Rogen, Hajime Kojima, Setsuya Aiba

Research output: Contribution to journalArticlepeer-review

Abstract

To evaluate the immunotoxic effects of xenobiotics, we have established the Multi-ImmunoTox assay, in which three stable reporter cell lines are used to evaluate the effects of chemicals on the IL-2, IFN-γ, IL-1β and IL-8 promoters. Here, we report the official validation study of the IL-2 luciferase assay (IL-2 Luc assay). In the Phase I study that evaluated five coded chemicals in three sets of experiments, the average within-laboratory reproducibility was 86.7%. In the Phase II study, 20 coded chemicals were evaluated at multiple laboratories. In the combined results of the Phase I and II studies, the between-laboratory reproducibility was 80.0%. These results suggested that the IL-2 Luc assay was reproducible both between and within laboratories. To determine the predictivity, we collected immunotoxicological information and constructed the reference data by classifying the chemical into immunotoxic compounds targeting T cells or others according to previously reported criteria. When compared with the reference data, the average predictivity of the Phase I and II studies was 75.0%, while that of additional 60 chemicals examined by the lead laboratory was 82.5%. Although the IL-2 Luc assay alone is not sufficient to predict immunotoxicity, it will be a useful tool when combined with other immune tests.

Original languageEnglish
Article number104832
JournalToxicology in Vitro
Volume66
DOIs
Publication statusPublished - 2020 Aug

Keywords

  • IL-2
  • Immunotoxic assay
  • Luciferase assay
  • Validation study

ASJC Scopus subject areas

  • Toxicology

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