Proteomics analysis of dendritic cell activation by contact allergens reveals possible biomarkers regulated by Nrf2.
Allergic contact dermatitis is a widespread disease with high clinical relevance affecting approximately 20% of the general population. Typically, contact allergens are low molecular weight electrophilic compounds which can activate the Keap1/Nrf2 pathway. We performed a proteomics study to reveal possible biomarkers for dendritic cell (DC) activation by contact allergens and to further elucidate the role of Keap1/Nrf2 signaling in this process. We used bone marrow derived dendritic cells (BMDCs) of wild-type (nrf2+/+) and Nrf2 knockout (nrf2-/-) mice and studied their response against the model contact sensitizers 2,4-dinitrochlorobenzene (DNCB), cinnamaldehyde (CA) and nickel(II) sulfate by 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) in combination with electrospray ionization tandem mass spectrometry (ESI-MS/MS). Sodium dodecyl sulfate (SDS, 100μM) served as irritant control. While treatment with nickel(II) sulfate and SDS had only little effects, CA and DNCB led to significant changes in protein expression. We found 18 and 30 protein spots up-regulated in wild-type cells treated with 50 and 100μM CA, respectively. For 5 and 10μM DNCB, 32 and 37 spots were up-regulated, respectively. Almost all of these proteins were not differentially expressed in nrf2-/-BMDCs, indicating an Nrf2-dependent regulation. Among them proteins were detected which are involved in oxidative stress and heat shock responses, as well as in signal transduction or basic cellular pathways. The applied approach allowed us to differentiate between Nrf2-dependent and Nrf2-independent cellular biomarkers differentially regulated upon allergen-induced DC activation. The data presented might contribute to the further development of suitable in vitro testing methods for chemical-mediated sensitization.
German Federal Institute for Risk Assessment (BfR), Department of Chemical and Product Safety, Berlin, Germany. Electronic address: franz.mussotter@bfr.bund.de.
Franz
F
Mussotter
Helmholtz Centre for Environmental Research (UFZ), Department of Molecular Systems Biology, Leipzig, Germany.
Janina Melanie
JM
Tomm
INSERM UMR 996, Univ Paris-Sud, Université Paris-Saclay, Chátenay-Malabry, France.
Zeina
Z
El Ali
INSERM UMR 996, Univ Paris-Sud, Université Paris-Saclay, Chátenay-Malabry, France.
Marc
M
Pallardy
INSERM UMR 996, Univ Paris-Sud, Université Paris-Saclay, Chátenay-Malabry, France.
Saadia
S
Kerdine-Römer
German Federal Institute for Risk Assessment (BfR), Department of Chemical and Product Safety, Berlin, Germany.
Mario
M
Götz
Helmholtz Centre for Environmental Research (UFZ), Department of Molecular Systems Biology, Leipzig, Germany; University of Leipzig, Institute of Biochemistry, Leipzig, Germany; Aalborg University, Department of Chemistry and Bioscience, Aalborg, Denmark.
Martin
M
von Bergen
German Federal Institute for Risk Assessment (BfR), Department of Chemical and Product Safety, Berlin, Germany.
Andrea
A
Haase
German Federal Institute for Risk Assessment (BfR), Department of Chemical and Product Safety, Berlin, Germany.
Andreas
A
Luch
D000485:Allergens / Q000494:pharmacology*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D003713:Dendritic Cells / Q000187:drug effects*; D051379:Mice; D018345:Mice, Knockout; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000502:physiology*; D040901:Proteomics
D000485:Allergens; D015415:Biomarkers; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse
2D-PAGE; Allergic contact dermatitis; Dendritic cell; Keap1/Nrf2; Proteomics; Sensitization