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J Appl Toxicol. 2016 Sep;36(9):1129-36. doi: 10.1002/jat.3274. Epub 2015 Dec 22.

Discrimination of skin sensitizers from non-sensitizers by interleukin-1α and interleukin-6 production on cultured human keratinocytes.

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Department of Microbiology and Immunology, and Institute of Endemic Disease, College of Medicine, Seoul National University, Seoul, 110-799, South Korea.
Biomedical Research Institute, Seoul National University Hospital, Seoul, 110-744, Republic of Korea.
College of Pharmacology, Ewha Womans University, Seoul, 120-808, Republic of Korea.
Chung-Ang University, College of Pharmacy, Seoul, 156-756, Republic of Korea.
Department of Occupational Health, College of Natural Sciences, Catholic University of Daegu, Daegu, 712-702, South Korea.


In vitro testing methods for classifying sensitizers could be valuable alternatives to in vivo sensitization testing using animal models, such as the murine local lymph node assay (LLNA) and the guinea pig maximization test (GMT), but there remains a need for in vitro methods that are more accurate and simpler to distinguish skin sensitizers from non-sensitizers. Thus, the aim of our study was to establish an in vitro assay as a screening tool for detecting skin sensitizers using the human keratinocyte cell line, HaCaT. HaCaT cells were exposed to 16 relevant skin sensitizers and 6 skin non-sensitizers. The highest dose used was the dose causing 75% cell viability (CV75) that we determined by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The levels of extracellular production of interleukin-1α (IL-1α) and IL-6 were measured. The sensitivity of IL-1α was 63%, specificity was 83% and accuracy was 68%. In the case of IL-6, sensitivity: 69%, specificity: 83% and accuracy: 73%. Thus, this study suggests that measuring extracellular production of pro-inflammatory cytokines IL-1α and IL-6 by human HaCaT cells may potentially classify skin sensitizers from non-sensitizers.


HaCaT; cultured human keratinocyte; interleukin-1α; interleukin-6; skin sensitizer

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