Role of oxidative stress in ERK and p38 MAPK activation induced by the chemical sensitizer DNFB in a fetal skin dendritic cell line

Abstract

SummaryThe intracellular mechanisms involved in the early phase of dendritic cell (DC) activation upon contact with chemical sensitizers are not well known. The strong skin sensitizer 2,4-dinitrofluorobenzene (DNFB) was shown to induce the activation of mitogen-activated protein kinases (MAPK) in DC. In the present study, we investigated a putative role for oxidative stress in DNFB-induced MAPK activation and upregulation of the costimulatory molecule CD40. In a DC line generated from fetal mouse skin, DNFB induced a significant increase in protein oxidation, measured by the formation of carbonyl groups, while it had almost no effect on lipid peroxidation. The antioxidants glutathione and vitamin E, which inhibit protein and lipid oxidation, respectively, were used to assess the role of oxidative stress in DNFB-induced MAPK activation. Glutathione, but not vitamin E, inhibited DNFB-induced p38 MAPK and ERK1/2 phosphorylation, whereas none of the antioxidants interfered significantly with the DNFB-induced upregulation of CD40 protein levels. Taken together, these results indicate that DNFB activates p38 MAPK and ERK1/2 via production of reactive oxygen species, and that protein oxidation plays an important role in MAPK activation.

Signal transduction profile of chemical sensitisers in dendritic cells: An endpoint to be included in a cell-based in vitro alternative approach to hazard identification?

“…TNF-α production induced by DNCB was also linked to the activation of p38 MAPK and JNK (Boisleve et al, 2004). In previous results performed with the foetal skin-derived dendritic cell line (FSDC), we reported that DNFB activates p38 MAPK and ERK1/2 via the production of reactive oxygen species and that protein oxidation plays an important role in MAPKs activation (Matos et al, 2005b). These results are in agreement with the key role of oxidative stress in the activation of MAPKs (Mizuashi et al, 2005;Hirota et al, 2009;Suzuki et al, 2009;Kagatani et al, 2010).…”

“…Accordingly, DNFB induced a significant increase in protein oxidation, as measured by the formation of carbonyl groups, while it had almost no effect on lipid peroxidation in the FSDC line. In addition, glutathione, but not vitamin E, inhibited DNFB-induced p38 MAPK and ERK1/2 phosphorylation (Matos et al, 2005b). Whether sensitisers induce redox or oxidative stress in dendritic cells was investigated by measuring the ratio of the oxidised (GSSG) versus the reduced (GSH) form of cellular glutathione (Mizuashi et al, 2005); all of the sensitisers (nickel, formaldehyde, DNCB, MnCl 2 , and thimerosal), but none of the non-sensitisers (SDS, BC, and ZnCl 2 ), reduced the GSH/GSSG ratio, which was accompanied by p38 MAPK phosphorylation.…”

Effect of lipopolysaccharide, skin sensitizers and irritants on thioredoxin-1 expression in dendritic cells: relevance of different signalling pathways

“…It was also demonstrated that the redox imbalance induced by skin sensitizers is important in the regulation of DC functions [5,38,39,41,47,61]. Therefore, the increase in Trx-1 mRNA expression observed in .…”

“…Similarly to LPS, skin sensitizers can induce redox imbalance [38,41] that activates the cellular mechanism proposed above and responsible for the increase in Trx-1 protein levels. These data suggest that the ASK1/Nrf2/ ARE regulatory pathway could be activated by LPS and skin sensitizers in a similar way, being involved in DC maturation.…”

Differential expression of cell surface markers in response to 2,4-dinitrofluorobenzene in RAW 264.7 and primary immune cells

“…DCs upregulate expression of MHC class II molecules and costimulatory molecules such as CD40, CD54, CD83, and CD86 in vitro in response to contact sensitizers (9,(11)(12)(13). DNFB mediates production of reactive oxygen species (ROS) and activates p38 mitogen-activated protein kinase (MAPK) and ERK1/2 to regulate CD40 expression (14,15). However, exposure of DCs to nickel sulfate stimulates signal transduction pathways and augments expression of MHC class II and several costimulatory molecules in different manners compared to that of DNFB (16).…”