Journal of Clinical & Experimental Dermatology Research
Open Access
ISSN: 2155-9554
+44 1478 350008
ISSN: 2155-9554
+44 1478 350008
Maria Teresa de Teixeira Cruz, Martins J D, Silva A, Goncalo M, Custodio J B A, Ferreira I, Domingues M R M, Lopes M C and Neves B M
University of Coimbra, Portugal
University of Aveiro, Portugal
Posters & Accepted Abstracts: Clin Exp Dermatol Res
Several reports link purinergic signaling to allergic contact dermatitis but the molecular mechanisms behind nucleotides signaling in the presence of skin allergens remain elusive. In the present work we have explored the possible involvement of purinergic signaling in the maturation process of dendritic like cells (THP-1) exposed to the skin sensitizer 1-fluoro-2,4-dinitrobenzene (DNFB). Due to their abundance in epidermis keratinocytes represent a relevant source of released purines following skin exposure to allergens. Therefore, we have used THP-1 co-cultured with the human keratinocyte cell line HaCaT to explore possible modulations of THP-1 cells and extracellular nucleotides by keratinocytes. Using high performance liquid chromatography we have determined the degradation pattern of ATP added to THP-1 cultures. We next compared the capacity of the different detected ATP metabolites to induce the expression of several DC maturation markers by quantitative PCR. Using pharmacological inhibitors we explored the involvement of purinergic receptors and membrane transporters in THP-1 cells maturation triggered by DNFB, cultured alone or co-cultured with keratinocytes. Since both skin sensitizers� and extracellular nucleotides� cellular actions rely on signaling by mitogen-activated protein kinases (MAPK), we further disclosed the putative crosstalk between DNFB and adenine nucleotides at the level of these signaling cascades. Overall, we found that purinergic signaling is involved in DNFB effects and that ADP, acting through metabotropic P2Y purinergic receptors is most probably the extracellular nucleotide involved. We did not find overlapping between DNFB and extracellular nucleotides effects at the level of MAPK signaling, suggesting that distinct upstream modulators of these cascades are involved.
Email: trosete@ff.uc.pt