Development of a novel in vitro assay to evaluate environmental water using an IL-8 reporter cell line

Authors

  • Yutaka Kimura Department of Dermatology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8574, Japan
  • Chizu Fujimura Department of Dermatology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8574, Japan
  • Toshifumi Imagawa Department of Virology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8575, Japan
  • Socorro P. Lupisan Research Institute for Tropical Medicine, FCC, Alabang, Muntinlupa 1781, Philippines
  • Mariko Saito-Obata Department of Virology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8575, Japan
  • Mayuko Saito Department of Virology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8575, Japan
  • Hitoshi Oshitani Department of Virology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8575, Japan
  • Setsuya Aiba Department of Dermatology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-cho, Aoba-ku, Sendai 980-8574, Japan, Tel: +81-22-717-7271; Fax: +81-22-717-7361, E-mail: saiba@med.tohoku.ac.jp

DOI:

https://doi.org/10.17179/excli2020-2104

Keywords:

Toll-like receptors, NOD-like receptors, IL-8, Luciferase assay, monocyte, hapten

Abstract

The IL-8 luciferase reporter cell line, THP-G8 cells, used in the in vitro sensitization test, OECD442E, can respond to a variety of stimuli other than haptens, such as lipopolysaccharide (LPS), other bacterial toxins, and detergents. Considering these characteristics, we examined the ability of the IL-8 luciferase assay using THP-G8 cells to evaluate water pollution. We first stimulated THP-G8 cell with various Toll-like receptor (TLR) agonists and nucleotide-binding oligomerization domain-like receptor (NLR) agonists, and found that TLR1, 2, 4, 5, 6 agonists and NOD 1, 2 agonists significantly augmented IL-8 luciferase activity (IL8LA). Then, we examined the detection threshold of LPS by THP-G8 cells, and found it 0.4 EU/ml. Next, we examined whether THP-G8 cells can differently respond to a variety of sources of environmental water around Sendai, Japan and Manila, Philippine and whether there is a correlation between the IL8LA of different sources of water and their level of endotoxin assessed by the LAL assay. There was a clear trend that the IL8LA was lower in the upper stream and higher in the downstream in both Japan and Philippine. Moreover, there was a strong correlation between the IL8LA of the environmental water and its endotoxin level. Finally, using N-acetyl-L-cysteine, an antioxidant/radical scavenger, and polymyxin B that neutralizes endotoxin, we demonstrated that there was a difference in the suppressive effects by them between the water from Japan and that from Philippine. These data suggest the potential of the IL-8 luciferase assay for evaluating environmental water pollution both quantitatively and qualitatively.

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Published

2020-07-31

How to Cite

Kimura, Y., Fujimura, C., Imagawa, T., Lupisan, S. P., Saito-Obata, M., Saito, M., … Aiba, S. (2020). Development of a novel in vitro assay to evaluate environmental water using an IL-8 reporter cell line. EXCLI Journal, 19, 1054–1063. https://doi.org/10.17179/excli2020-2104

Issue

Vol. 19 (2020)

Section

Original articles

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Copyright (c) 2020 Yutaka Kimura, Chizu Fujimura, Toshifumi Imagawa, Socorro P. Lupisan, Mariko Saito-Obata, Mayuko Saito, Hitoshi Oshitani, Setsuya Aiba

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