TY - JOUR
T1 - Development of a novel in vitro assay to evaluate environmental water using an IL-8 reporter cell line
AU - Kimura, Yutaka
AU - Fujimura, Chizu
AU - Imagawa, Toshifumi
AU - Lupisan, Socorro P.
AU - Saito-Obata, Mariko
AU - Saito, Mayuko
AU - Oshitani, Hitoshi
AU - Aiba, Setsuya
N1 - Funding Information:
This work was supported by JSPS KA- KENHI (Grant-in-Aid for challenging Exploratory Research, 25550076), Grants-in-Aid from the Ministry of Health, Labor and Welfare (MHLW, 0618010), and Japan Initiative for Global Research Network from Japan Agency for Medical Research and Development (JP19fm010813 and JPwm0125001).
Publisher Copyright:
© 2020, Leibniz Research Centre for Working Environment and Human Factors. All rights reserved.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - Hapten
KW - IL-8
KW - Luciferase assay
KW - Monocyte
KW - NOD-like receptors
KW - Toll-like receptors
UR - http://www.scopus.com/inward/record.url?scp=85089625529&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85089625529&partnerID=8YFLogxK
U2 - 10.17179/excli2020-2104
DO - 10.17179/excli2020-2104
M3 - Article
AN - SCOPUS:85089625529
VL - 19
SP - 1054
EP - 1063
JO - EXCLI Journal
JF - EXCLI Journal
SN - 1611-2156
ER -