TY - JOUR
T1 - Evaluation of changes of cell-surface thiols as a new biomarker for in vitro sensitization test
AU - Suzuki, Mie
AU - Hirota, Morihiko
AU - Hagino, Shigenobu
AU - Itagaki, Hiroshi
AU - Aiba, Setsuya
PY - 2009/6
Y1 - 2009/6
N2 - In order to find a novel biomarker for a simple assay to predict skin sensitization, we evaluated cell-surface thiols as a biomarker reflecting intracellular signaling in THP-1 cells (human monocytic cell line). First, we found that a decrease of cell-surface thiols on hapten-treated THP-1 cells was induced in parallel with phosphorylation of p38 MAPK. Next, we confirmed that 2-mercaptoethanol in the culture medium and the differentiation state of THP-1 cells did not affect the changes of cell-surface thiols by hapten. Changes of cell-surface thiols on THP-1 cells were detected after 2 h treatment with most allergens (e.g., DNCB, NiSO4), as well as some non-allergens (e.g., Tween80, benzalkonium chloride), though other non-allergens (e.g., SDS, glycerol) had no effect. When either a significant decrease or increase of cell-surface thiols (more than 15% in each case) was used as a criterion, the results using 36 allergens and 16 non-allergens were in good accordance with those of in vivo assays. Finally, we confirmed that ATP, which is released as a consequence of cytotoxicity, did not affect the changes of cell-surface thiols. Our results suggest that changes of cell-surface thiols may be useful for an in vitro sensitization assay, which we designate as the SH test.
AB - In order to find a novel biomarker for a simple assay to predict skin sensitization, we evaluated cell-surface thiols as a biomarker reflecting intracellular signaling in THP-1 cells (human monocytic cell line). First, we found that a decrease of cell-surface thiols on hapten-treated THP-1 cells was induced in parallel with phosphorylation of p38 MAPK. Next, we confirmed that 2-mercaptoethanol in the culture medium and the differentiation state of THP-1 cells did not affect the changes of cell-surface thiols by hapten. Changes of cell-surface thiols on THP-1 cells were detected after 2 h treatment with most allergens (e.g., DNCB, NiSO4), as well as some non-allergens (e.g., Tween80, benzalkonium chloride), though other non-allergens (e.g., SDS, glycerol) had no effect. When either a significant decrease or increase of cell-surface thiols (more than 15% in each case) was used as a criterion, the results using 36 allergens and 16 non-allergens were in good accordance with those of in vivo assays. Finally, we confirmed that ATP, which is released as a consequence of cytotoxicity, did not affect the changes of cell-surface thiols. Our results suggest that changes of cell-surface thiols may be useful for an in vitro sensitization assay, which we designate as the SH test.
KW - Cell-surface thiols
KW - In vitro
KW - SH test
KW - Skin sensitization
KW - THP-1 cells
KW - p38 MAPK
UR - http://www.scopus.com/inward/record.url?scp=66149178433&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=66149178433&partnerID=8YFLogxK
U2 - 10.1016/j.tiv.2009.02.002
DO - 10.1016/j.tiv.2009.02.002
M3 - Article
C2 - 19490838
AN - SCOPUS:66149178433
VL - 23
SP - 687
EP - 696
JO - Toxicology in Vitro
JF - Toxicology in Vitro
SN - 0887-2333
IS - 4
ER -