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Understanding the skin sensitization capacity of ascaridole: a combined study of chemical reactivity and activation of the innate immune system (dendritic cells) in the epidermal environment

  • Fatma Sahli
  • Marta Silva E. Sousa
  • Bertrand Vileno
  • Jutta Lichter
  • Jean-Pierre Lepoittevin
  • Brunhilde Blömeke
  • Elena Giménez-ArnauEmail author
Immunotoxicology

Abstract

To improve the prediction of the possible allergenicity of chemicals in contact with the skin, investigations of upstream events are required to better understand the molecular mechanisms involved in the initiation of allergic reactions. Ascaridole, one of the compounds responsible for skin sensitization to aged tea tree oil, degrades into intermediates that evolve via different mechanisms involving radical species. We aimed at broadening the knowledge about the contribution of radical intermediates derived from ascaridole to the skin sensitization process by assessing the reactivity profile towards amino acids, identifying whether free radicals are formed in a reconstructed human epidermis (RHE) model and their biological properties to activate the immune system, namely dendritic cells in their natural context of human HaCaT keratinocytes and RHE. Electron paramagnetic resonance combined to spin-trapping in EpiSkin™ RHE confirmed the formation of C-radicals in the epidermal tissue from 10 mM ascaridole concentration, while reactivity studies toward amino acids showed electrophilic intermediates issued from radical rearrangements of ascaridole as the main reactive species. Activation of THP-1 cells, as surrogate for dendritic cells, that were cocultured with HaCaT was significantly upregulated after treatment with low micromolar concentrations based on cell surface expression of the co-stimulatory molecule CD86 and the adhesion molecule CD54. Placing THP-1 cells underneath the RHE allowed us to monitor which of the concentrations that produce radical(s) and/or protein antigens in the epidermal skin environment promote the activation of dendritic cells. We detected no significant upregulation of CD86/CD54 after topical RHE application of concentrations up to 30 mM ascaridole (t = 24 h) but clear upregulation after 60 mM.

Keywords

Skin sensitization Dendritic cell activation THP-1 cells Keratinocytes Ascaridole Reactivity EPR-spectroscopy Spin-trapping Reconstructed human epidermis 

Notes

Acknowledgements

This work was part of the DEFCHEMSKALL Franco-German Collaborative International Research Project financially supported by the ANR (Agence Nationale de la Recherche; project no. ANR-15-CE15-0023-01) and the DFG (Deutsche Forschungsgemeinschaft; project no. DFG, BL340/6-1). The Reseau NAtional de Rpe interDisciplinaire (RENARD, Fédération IR-RPE CNRS #3443) is also acknowledged.

Compliance with ethical standards

The manuscript does not contain clinical studies or patient data.

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

204_2019_2444_MOESM1_ESM.pdf (596 kb)
Supplementary material 1 (PDF 595 kb)

References

  1. Barbati S, Clément JL, Olive G, Roubaud V, Tuccio B, Tordo P (1997) In: Minisci F (ed) Free radicals in biology and environment. Kluwer Academic Publishers, Dordrecht, pp 39–47CrossRefGoogle Scholar
  2. Bartesaghi S, Radi R (2018) Fundamentals on the biochemistry of peroxynitrite and protein tyrosine nitration. Redox Biol 14:618–625.  https://doi.org/10.1016/j.redox.2017.09.009 CrossRefGoogle Scholar
  3. Bocchietto E, Paolucci C, Breda D, Sabbioni E, Burastero SE (2007) Human monocytoid THP-1 cell line versus monocyte-derived human immature dendritic cells as in vitro models for predicting the sensitizing potential of chemicals. Int J Immunopathol Pharmacol 20:259–265.  https://doi.org/10.1177/039463200702000206 CrossRefGoogle Scholar
  4. Carroll L, Pattison DI, Davies JB, Anderson RF, Lopez-Alarcon C, Davies MJ (2018) Superoxide radicals react with peptide-derived tryptophan radicals with very high rate constants to give hydroperoxydes as major products. Free Radic Biol Med 118:126–136.  https://doi.org/10.1016/j.freeradbiomed.2018.02.033 CrossRefGoogle Scholar
  5. Chittiboyina AG, Avonto C, Khan IA (2016) What happens after activation of ascaridole? Reactive compounds and their implications for skin sensitization. Chem Res Toxicol 29:1488–1492.  https://doi.org/10.1021/acs.chemrestox.6b00157 CrossRefGoogle Scholar
  6. Christoffers WA, Blömeke B, Coenraads P-J, Schuttelar M-LA (2013) Co-sensitization to ascaridole and tea tree oil. Contact Dermatitis 69:181–191.  https://doi.org/10.1111/cod.12086 CrossRefGoogle Scholar
  7. Christoffers WA, Blömeke B, Coenraads P-J, Schuttelar M-LA (2014) The optimal patch test concentration for ascaridole as a sensitizing component of tea tree oil. Contact Dermatitis 71:129–137.  https://doi.org/10.1111/cod.12199 CrossRefGoogle Scholar
  8. Clément JL, Finet JP, Fréjaville C, Tordo P (2003) Deuterated analogues of the free radical trap DEPMPO: synthesis and EPR studies. Org Biomol Chem 1:1591–1597.  https://doi.org/10.1039/B300870C CrossRefGoogle Scholar
  9. de Groot AC, Schmidt E (2016) Tea tree oil: contact allergy and chemical composition. Contact Dermatitis 75:129–143.  https://doi.org/10.1111/cod.12591 CrossRefGoogle Scholar
  10. Frejaville C, Karoui H, Tuccio B, Le Moigne F, Culcasi M, Pietri S, Lauricella R, Tordo P (1995) 5-(Diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide: a new efficient phosphorylated nitrone for the in vitro spin trapping of oxygen-centered radicals. J Med Chem 38:258–265.  https://doi.org/10.1021/jm00002a007 CrossRefGoogle Scholar
  11. Fujimoto A, Masuda T (2012) Chemical interaction between polyphenols and a cysteinyl thiol under radical oxidation controls. J Agric Food Chem 60:5142–5151.  https://doi.org/10.1021/jf3008822 CrossRefGoogle Scholar
  12. Geroldinger G, Tonner M, Hettegger H, Bacher M, Monzote L, Walter M, Staniek K, Rosenau T, Gille L (2017) Mechanism of ascaridole activation in Leishmania. Biochem Pharmacol 132:48–62.  https://doi.org/10.1016/j.bcp.2017.02.023 CrossRefGoogle Scholar
  13. Hausen BM, Reichling J, Harkenthal M (1999) Degradation products of monoterpenes are the sensitizing agents in tea tree oil. Am J Contact Dermatitis 10:68–77.  https://doi.org/10.1016/S1046-199X(99)90002-7 CrossRefGoogle Scholar
  14. Hennen J, Bloemeke B (2017) COCAT-advanced in vitro assessment of skin sensitization potency of chemicals using THP-1 cells in coculture with HaCaT keratinocytes. In: Naunyn-Schmiedebergs Archives of Pharmacology. Vol 390. Springer, 233 Spring St, New York, NY 10013 USA, pp S87–S87Google Scholar
  15. Hennen J, Aeby P, Goebel C et al (2011) Cross talk between keratinocytes and dendritic cells: impact on the prediction of sensitization. Toxicol Sci 123:501–510.  https://doi.org/10.1093/toxsci/kfr174 CrossRefGoogle Scholar
  16. Kao D, Chaintreau A, Lepoittevin J-P, Giménez-Arnau E (2011) Synthesis of allylic hydroperoxides and EPR spin-trapping studies on the formation of radicals in iron systems as potential initiators of the sensitizing pathway. J Org Chem 76:6188–6200.  https://doi.org/10.1021/jo200948x CrossRefGoogle Scholar
  17. Kao D, Chaintreau A, Lepoittevin J-P, Giménez-Arnau E (2014) Mechanistic studies on the reactivity of sensitizing allylic hydroperoxides: investigation of the covalent modification of amino acids by carbon-radical intermediates. Toxicol Res 3:278–289.  https://doi.org/10.1039/C3TX50109D CrossRefGoogle Scholar
  18. Krutz NL, Hennen J, Korb C, Schellenberger MT, Gerberick GF, Blömeke B (2015) Activation of the endoperoxide ascaridole modulates its sensitizing capacity. Toxicol Sci 147:515–523.  https://doi.org/10.1093/toxsci/kfv148 CrossRefGoogle Scholar
  19. Kuresepi S, Vileno B, Turek P, Lepoittevin J-P, Giménez-Arnau E (2018) Potential of EPR spin-trapping to investigate in situ free radicals generation from skin allergens in reconstructed human epidermis: cumene hydroperoxyde as proof of concept. Free Radic Res 52:171–179.  https://doi.org/10.1080/10715762.2017.1420906 CrossRefGoogle Scholar
  20. Lauricella R, Tuccio B (2014) Détection et caractérisation de radicaux libres après piégeage de spins. In: Bertrand P (ed) La Spectroscopie de Résonance Paramagnétique Électronique-Applications. Collection Grenoble Sciences, edp Sciences, pp 49–78Google Scholar
  21. Lepoittevin JP (2011) Molecular aspects in allergic and irritant contact dermatitis. In: Johansen JD, Frosch PJ, Lepoittevin JP (eds) Contact dermatitis, 5th edn. Springer, Berlin, pp 91–110CrossRefGoogle Scholar
  22. Monzote L, Stamberg W, Staniek K, Gille L (2009) Toxic effects of carvacrol, caryophyllene oxide, and ascaridole from essential oil of Chenopodium ambrosioides on mithocondria. Toxicol Appl Pharmacol 240:337–347.  https://doi.org/10.1016/j.taap.2009.08.001 CrossRefGoogle Scholar
  23. Mori HM, Iwahashi H (2013) Detection and identification of 1-methylethyl and methyl radicals generated by irradiating tea tree (Melaleuca alternifolia) oil with visible light (436 nm) in the presence of flavin mononucleotide and ferrous ion. Free Radic Res 47:657–663.  https://doi.org/10.3109/10715762.2013.812785 CrossRefGoogle Scholar
  24. O’Neill PM, Barton VE, Ward SA (2010) The molecular mechanism of action of artemisinin-the debate continues. Molecules 15:1705–1721.  https://doi.org/10.3390/molecules15031705 CrossRefGoogle Scholar
  25. OECD (2012) The adverse outcome pathway for skin sensitization initiated by covalent binding to proteins. Part 1: Scientific evidence. OECD Series on Testing and Assessment No. 168 (ENV/JM/MONO(2012)10/PART1). OECD Publishing, Paris.  https://doi.org/10.1787/9789264221444-en
  26. OECD testing guideline 439 (2015) In vitro skin irritation: RHE test method.  https://doi.org/10.1787/9789264242845-en
  27. Pazos M, Andersen ML, Skibsted LH (2006) Amino acid and protein scavenging of radicals generated by iron/hydroperoxide system: an electron spin resonance spin trapping study. J Agric Food Chem 54:10215–10221.  https://doi.org/10.1021/jf062134n CrossRefGoogle Scholar
  28. Pesonen M, Jolanki R, Larese Filon F, Wilkinson M, Kręcisz B, Kieć-Swierczyńska M et al (2015) Patch test results of the European baseline series among patients with occupational contact dermatitis across Europe—analyses of the European Surveillance System on Contact Allergy network, 2002–2010. Contact Dermatitis 72:154–163.  https://doi.org/10.1111/cod.12333 CrossRefGoogle Scholar
  29. Rudbäck J, Andresen Bergström M, Börje A, Nilsson U, Karlberg A-T (2012) α-Terpinene, an antioxidant in tea tree oil, autoxidizes rapidly to skin allergens on air exposure. Chem Res Toxicol 25:713–721.  https://doi.org/10.1021/tx200486f CrossRefGoogle Scholar
  30. Satyal P, Paudel P, Kafme A, Pokharel SK, Lamichhane B, Dosoky NS et al (2012) Bioactivities of volatile components from Nepalese Artemisia species. Nat Prod Commun 12:1651–1658Google Scholar
  31. Scientific Committee on Consumer Products (SCCP) (2008) Opinion on Tea Tree Oil. Adopted by the SCCP during the 18th plenary meeting of 16 December 2008. SCCP Report 1155/08. https://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_160.pdf
  32. Steinman RM (2007) Dendritic cells: understanding immunogenicity. Eur J Immunol 37(Suppl 1):S53–S60.  https://doi.org/10.1002/eji.200737400 CrossRefGoogle Scholar
  33. Stoll S, Schweiger A (2006) EasySpin, a comprehensive software package for spectral simulation and analysis in EPR. J Magn Res 178:42–55.  https://doi.org/10.1016/j.jmr.2005.08.013 CrossRefGoogle Scholar
  34. Stolz K, Udilova N, Nohl H (2000) Spin trapping of lipid radicals with DEPMPO-derived spin traps: detection of superoxide, alkyl and alkoxyl radicals in aqueous and lipid phase. Free Radic Biol Med 29:1005–1014.  https://doi.org/10.1016/S0891-5849(00)00401-9 CrossRefGoogle Scholar
  35. Tang Y, Dong Y, Wang X, Sriraghavan K, Wood JK, Vennerstrom JL (2005) Dispiro-1,2,4-trioxane analogues of a prototype dispiro-1,2,4-trioxolane: mechanistic comparators for artemisinin in the context of reaction pathways with iron (II). J Org Chem 70:5103–5110.  https://doi.org/10.1021/jo050385+ CrossRefGoogle Scholar
  36. Tietze C, Blömeke B (2008) Sensitization assays: monocyte-derived dendritic cells versus a monocytic cell line (THP-1). J Toxicol Environ Health A 71:965–968.  https://doi.org/10.1080/15287390801989168 CrossRefGoogle Scholar
  37. Timmins GS, Davies MJ (1993) Free radical formation in murine skin treated with tumour promoting organic peroxides. Carcinogenesis 14:1499–1503.  https://doi.org/10.1093/carcin/14.8.1499 CrossRefGoogle Scholar
  38. Wang D-Y, Wu Y-L, Wu Y, Liang J, Li Y (2001) Further evidence for the participation of primary carbon-centered free radicals in the antimalarial action of the qinghaosu (artemisinin) series of compounds. J Chem Soc Perkin Trans 1:605–609.  https://doi.org/10.1039/b008145k CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Dermatochemistry LaboratoryUniversity of Strasbourg, CNRS, UMR 7177, Institut le BelStrasbourgFrance
  2. 2.Department of Environmental ToxicologyUniversity TrierTrierGermany
  3. 3.POMAM LaboratoryUniversity of Strasbourg, CNRS, UMR 7177, Institut le BelStrasbourgFrance
  4. 4.French EPR Federation of Research, REseau NAtional de Rpe interDisciplinaire, RENARDStrasbourgFrance

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