ToxSci Advance Access originally published online on November 25, 2008
Toxicological Sciences 2009 107(2):451-460; doi:10.1093/toxsci/kfn243
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HMOX1 and NQO1 Genes are Upregulated in Response to Contact Sensitizers in Dendritic Cells and THP-1 Cell Line: Role of the Keap1/Nrf2 Pathway
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* University of Paris-Sud, INSERM UMR-S 749, Faculté de Pharmacie Paris-Sud, 5, rue JB Clément, F-92296 Châtenay-Malabry
INSERM, Faculté de Pharmacie Paris-Sud, 5, rue JB Clément, F-92296 Châtenay-Malabry, France
AFSSAPS, DLC/BCM, 635 Rue de la garenne F-34740 Vendargues, France
1 To whom correspondence should be addressed at AFSSAPS, DLC/BCM, 635 Rue de la garenne F-34740 Vendargues, France. Fax: +33-4-67-91-39-82. E-mail: jean-claude.ourlin{at}afssaps.sante.fr.
Received July 31, 2008; accepted November 18, 2008
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Abstract |
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Electrophilicity is one of the most common features of skin contact sensitizers and is necessary for protein haptenation. The Keap1 (Kelch-like ECH-associated protein 1)/Nrf2 -signaling pathway is dedicated to the detection of electrophilic stress in cells leading to the upregulation of genes involved in protection or neutralization of chemical reactive species. Signals provided by chemical stress could play an important role in dendritic cell activation and the aim of this work was to test whether contact sensitizers were specific activators of the Keap1/Nrf2 pathway. CD34-derived dendritic cells (CD34-DC) and the THP-1 myeloid cell line were treated by a panel of sensitizers (Ni, 1-chloro 2,4-dinitrobenzene, cinnamaldehyde, 7-hydroxycitronellal, 1,4-dihydroquinone, 
-methyl-trans-cinnamaldehyde, 2-4-tert-(butylbenzyl)propionaldehyde or Lilial, and 1,4-phenylenediamine), irritants (sodium dodecyl sulfate, benzalkonium chloride), and a nonsensitizer molecule (chlorobenzene). Three well-known Nrf2 activators (tert-butylhydroquinone, lipoic acid, sulforaphane) were also tested. Expression of hmox1 and nqo1 was measured using real-time PCR and cellular accumulation of Nrf2 was assessed by Western blot. Our results showed an increased expression at early time points of hmox1 and nqo1 mRNAs in response to sensitizers but not to irritants. Accumulation of the Nrf2 protein was also observed only with chemical sensitizers. A significant inhibition of the expression of hmox1 and nqo1 mRNAs and CD86 expression was found in 1-chloro 2,4-dinitrobenzene–treated THP-1 cells preincubated with N-acetyl cysteine, a glutathione precursor. Altogether, these data suggested that the Keap1/Nrf2-signaling pathway was activated by electrophilic molecules including sensitizers in dendritic cells and in the THP-1 cell line. Monitoring of this pathway may provide new biomarkers (e.g., Nrf2, hmox1) for the detection of the sensitization potential of chemicals.
Key Words: skin sensitization; Nrf2; hmox1; oxidative stress.
Co-authors: Ade N. and Leon F. have contributed equally to this work and should be considered both as first authors.