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

Daun Jung, Jeong Hwan Che, Kyung Min Lim, Young Jin Chun, Yong Heo, Seung Hyeok Seok

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)1129-1136
Number of pages8
JournalJournal of Applied Toxicology
Volume36
Issue number9
DOIs
StatePublished - 1 Sep 2016

Keywords

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

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