Thumbnail
Access Restriction
Open

Author Emter, Roger ♦ Ellis, Graham ♦ Natsch, Andreas
Source United States Department of Energy Office of Scientific and Technical Information
Content type Text
Language English
Subject Keyword APPLIED LIFE SCIENCES ♦ IN VITRO ♦ LYMPH NODES ♦ PEPTIDES ♦ SENSITIZERS ♦ SKIN ♦ BODY ♦ LYMPHATIC SYSTEM ♦ ORGANIC COMPOUNDS ♦ ORGANS ♦ PROTEINS ♦ REAGENTS
Abstract In vitro tests are needed to replace animal tests to screen for the skin sensitization potential of chemicals. Skin sensitizers are electrophilic molecules and the Nrf2-electrophile-sensing pathway comprising the repressor protein Keap1, the transcription factor Nrf2 and the antioxidant response element (ARE) is emerging as a toxicity pathway induced by skin sensitizers. Previously, we screened a large set of chemicals in the reporter cell line AREc32, which contains an eight-fold repeat of the rat GSTA2 ARE-sequence upstream of a luciferase reporter gene in the human breast cancer cell line MCF7. This approach was now further developed to bring it closer to the conditions in the human skin and to propose a fully standardized assay. To this end, a luciferase reporter gene under control of a single copy of the ARE-element of the human AKR1C2 gene was stably inserted into HaCaT keratinocytes. A standard operating procedure was developed whereby chemicals are routinely tested at 12 concentrations in triplicate for significant induction of gene activity. We report results from this novel assay on (i) a list of reference chemicals published by ECVAM, (ii) the ICCVAM list of chemicals for validation of alternative endpoints in the LLNA and (iii) on a more general list of 67 chemicals derived from the ICCVAM database. For comparison, peptide reactivity data are presented for the same chemicals. The results indicate a good predictive value of this approach for hazard identification. Its technical simplicity, the high-throughput format and the good predictivity may make this assay a candidate for rapid validation to meet the tight deadline to replace animal tests for skin sensitization by 2013 set by the European authorities.
ISSN 0041008X
Educational Use Research
Learning Resource Type Article
Publisher Date 2010-06-15
Publisher Place United States
Journal Toxicology and Applied Pharmacology
Volume Number 245
Issue Number 3


Open content in new tab

   Open content in new tab