Skip to main content

Advertisement

SpringerLink
Log in

The THP-1 cell toolbox: a new concept integrating the key events of skin sensitization

  • Immunotoxicology
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

According to the current scientific consensus, one in vitro test is insufficient to cover the key events (KE) defined by the adverse outcome pathway (AOP) for skin sensitization. To address this issue we combined different end points in the same cell line to cover all KEs defined by the skin sensitization AOP. Since dendritic cells (DC) play a key role in the sensitization phase leading to the development of allergic contact dermatitis (ACD), we used THP-1 cells as a surrogate for DC. We measured ROS production and GSH depletion for KE1 (binding to proteins), Nrf2 activation pathway and gene expressions for KE2 (keratinocyte response), phenotype modifications using cell-surface markers and cytokine production for KE3 (DC activation), and T-cell proliferation for KE4 (T-cell activation). These measurements were performed using the THP-1 cell line and an original THP-1/T-cell co-culture system following exposure to a variety of chemicals, including irritant, non-sensitizers, and chemicals sensitizers (pro/prehaptens). Results showed that treatment with sensitizers such as cinnamaldehyde (100 µM) or methylisothiazolinone (150 µM) was able to trigger the three main key events (KE1, KE2, and KE3) of the sensitization phase of ACD in THP-1 cells. In addition, all sensitizers were able to induce T lymphocyte proliferation (KE4), while non-sensitizers and irritants did not. Our study shows for the first time that addressing the four main KE of skin sensitization AOP in a single cell line is an achievable task.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ACD:

Allergic contact dermatitis

AOP:

Adverse outcome pathway

ARE:

Antioxidant response element

BZK:

Benzalkonium chloride

CinA:

Cinnamaldehyde

CS:

Contact sensitizer

DC:

Dendritic cell

DNCB:

Dinitrochlorobenzene

DPRA:

Direct peptide reactivity assay

EUG:

Eugenol

FAR:

Farnesol

GER:

Geraniol

GSH:

Glutathione

GSSG:

Glutathione disulfide

h-CLAT:

human cell line activation test

ITS:

Integrated testing strategy

KE:

Key event

Keap1:

Kelch-like ECH-associated protein 1

MIT:

Methylisothiazolinone

Nrf2:

Nuclear factor erythroid 2-related factor 2

RESO:

Resorcinol

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulfate

t-BHQ:

tert-butylhydroquinone

References

  • Ade N, Antonios D, Kerdine-Romer S, Boisleve F, Rousset F, Pallardy M (2007) NF-kappaB plays a major role in the maturation of human dendritic cells induced by NiSO(4) but not by DNCB. Toxicol Sci 99:488–501

    Article  CAS  PubMed  Google Scholar 

  • Ade N, Leon F, Pallardy M, Peiffer J-L, Kerdine-Romer S, Tissier M-H, Bonnet P-A, Fabre I, Ourlin J-C (2009) 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. Toxicol Sci 107:451–460

    Article  CAS  PubMed  Google Scholar 

  • Ashikaga T, Sakaguchi H, Sono S, Kosaka N, Ishikawa M, Nukada Y, Miyazawa M, Ito Y, Nishiyama N, Itagaki H (2010) A comparative evaluation of in vitro skin sensitisation tests: the human cell-line activation test (h-CLAT) versus the local lymph node assay (LLNA). Altern Lab Anim 38:275–284

    Article  CAS  Google Scholar 

  • Aw Yeang HX, Hamdam JM, Al-Huseini LMA, Sethu S, Djouhri L, Walsh J, Kitteringham N, Park BK, Goldring CE, Sathish JG (2012) Loss of transcription factor nuclear factor-erythroid 2 (NF-E2) p45-related factor-2 (Nrf2) leads to dysregulation of immune functions, redox homeostasis, and intracellular signaling in dendritic cells. J Biol Chem 287:10556–10564

    Article  CAS  PubMed  Google Scholar 

  • Bechara R, Antonios D, Azouri H, Pallardy M (2017) Nickel sulfate promotes IL-17A producing cd4 + t cells by an il-23-dependent mechanism regulated by TLR4 and Jak-STAT pathways. J Invest Dermatol 137:2140–2148

    Article  CAS  PubMed  Google Scholar 

  • Boislève F, Kerdine-Römer S, Rougier-Larzat N, Pallardy M (2004) Nickel and DNCB induce CCR7 expression on human dendritic cells through different signalling pathways: Role of TNF- and MAPK. J Invest Dermatol 123:494–502

    Article  PubMed  Google Scholar 

  • Byamba D, Kim TG, Kim DH, Je JH, Lee M-G (2010) The roles of reactive oxygen species produced by contact allergens and irritants in monocyte-derived dendritic cells. Ann Dermatol 22:269–278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Casati S, Aeby P, Basketter DA, Cavani A, Gennari A, Gerberick GF, Griem P, Hartung T, Kimber I, Lepoittevin J-P, Meade BJ, Pallardy M, Rougier N, Rousset F, Rubinstenn G, Sallusto F, Verheyen GR, Zuang V (2005) Dendritic cells as a tool for the predictive identification of skin sensitisation hazard. Altern Lab Anim 33:47–62

    Article  CAS  Google Scholar 

  • Cottrez F, Boitel E, Auriault C, Aeby P, Groux H (2015) Genes specifically modulated in sensitized skins allow the detection of sensitizers in a reconstructed human skin model. Development of the SENS-IS assay. Toxicol Vitr 29:787–802

    Article  CAS  Google Scholar 

  • Cottrez F, Boitel E, Ourlin JC, Peiffer JL, Fabre I, Henaoui IS, Mari B, Vallauri A, Paquet A, Barbry P, Auriault C, Aeby P, Groux H (2016) SENS-IS, a 3D reconstituted epidermis based model for quantifying chemical sensitization potency: reproducibility and predictivity results from an inter-laboratory study. Toxicol Vitr 32:248–260

    Article  CAS  Google Scholar 

  • Cruikshank WW, Berman JS, Theodore AC, Bernardo J, Center DM (1987) Lymphokine activation of T4 + T lymphocytes and monocytes. J Immunol 138:3817–3823

    CAS  PubMed  Google Scholar 

  • de Bourayne M, Gallais Y, Ali E, Rousseau Z, Damiens P, Cochet M-H, Filhol C, Chollet-Martin O, Pallardy S, Kerdine-Romer M, S (2017) Protein kinase CK2 controls T-cell polarization through dendritic cell activation in response to contact sensitizers. J Leukoc Biol 101:703–715

    Article  PubMed  Google Scholar 

  • Dickinson DA, Levonen A-L, Moellering DR, Arnold EK, Zhang H, Darley-Usmar VM, Forman HJ (2004) Human glutamate cysteine ligase gene regulation through the electrophile response element. Free Radic Biol Med 37:1152–1159

    Article  CAS  PubMed  Google Scholar 

  • El Ali Z, Delomenie C, Botton J, Pallardy M, Kerdine-Romer S (2017) Dendritic cells’ death induced by contact sensitizers is controlled by Nrf2 and depends on glutathione levels. Toxicol Appl Pharmacol 322:41–50

    Article  CAS  PubMed  Google Scholar 

  • Emter R, Ellis G, Natsch A (2010) Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro. Toxicol Appl Pharmacol 245:281–290

    Article  CAS  PubMed  Google Scholar 

  • European Chemicals Agency (2016) Guidance on information requirements and chemical safety assessment. Chapter R7a: Endpoint specific guidance. European Chemicals Agency, Helsinki

    Google Scholar 

  • Franco R, Cidlowski JA (2012) Glutathione efflux and cell death. Antioxid Redox Signal 17:1694–1713

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gerberick GF, Vassallo JD, Bailey RE, Chaney JG, Morrall SW, Lepoittevin J-P (2004) Development of a peptide reactivity assay for screening contact allergens. Toxicol Sci 81:332–343

    Article  CAS  PubMed  Google Scholar 

  • Gerberick GF, Vassallo JD, Foertsch LM, Price BB, Chaney JG, Lepoittevin J-P (2007) Quantification of chemical peptide reactivity for screening contact allergens: a classification tree model approach. Toxicol Sci 97:417–427

    Article  CAS  PubMed  Google Scholar 

  • Hitzler M, Bergert A, Luch A, Peiser M (2013) Evaluation of selected biomarkers for the detection of chemical sensitization in human skin: a comparative study applying THP-1, MUTZ-3 and primary dendritic cells in culture. Toxicol In Vitro 27:1659–1669

    Article  CAS  PubMed  Google Scholar 

  • Kim ST, Takeuchi K, Sun Z-YJ, Touma M, Castro CE, Fahmy A, Lang MJ, Wagner G, Reinherz EL (2009) The alphabeta T cell receptor is an anisotropic mechanosensor. J Biol Chem 284:31028–31037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kimber I, Basketter DA, Gerberick GF, Ryan CA, Dearman RJ (2011) Chemical allergy: translating biology into hazard characterization. Toxicol Sci 120(Suppl):238–268

    Article  CAS  Google Scholar 

  • Lim Y-M, Moon S-J, An S-S, Lee S-J, Kim S-Y, Chang I-S, Park K-L, Kim H-A, Heo Y (2008) Suitability of macrophage inflammatory protein-1β production by THP-1 cells in differentiating skin sensitizers from irritant chemicals. Contact Dermatitis 58:193–198

    Article  CAS  PubMed  Google Scholar 

  • Loetscher P, Seitz M, Clark-Lewis I, Baggiolini M, Moser B (1994) Monocyte chemotactic proteins MCP-1, MCP-2, and MCP-3 are major attractants for human CD4 + and CD8 + T lymphocytes. FASEB J Off Publ Fed Am Soc Exp Biol 8:1055–1060

    CAS  Google Scholar 

  • Migdal C, Tailhardat M, Courtellemont P, Haftek M, Serres M (2010) Responsiveness of human monocyte-derived dendritic cells to thimerosal and mercury derivatives. Toxicol Appl Pharmacol 246:66–73

    Article  CAS  PubMed  Google Scholar 

  • Migdal C, Botton J, Ali E, Azoury Z, Guldemann M-E, Gimenez-Arnau J, Lepoittevin E, Kerdine-Romer J-P, Pallardy S, M (2013) Reactivity of chemical sensitizers toward amino acids in cellulo plays a role in the activation of the Nrf2-ARE pathway in human monocyte dendritic cells and the THP-1 cell line. Toxicol Sci 133:259–274

    Article  CAS  PubMed  Google Scholar 

  • Mizuashi M, Ohtani T, Nakagawa S, Aiba S (2005) Redox imbalance induced by contact sensitizers triggers the maturation of dendritic cells. J Invest Dermatol 124:579–586

    Article  CAS  PubMed  Google Scholar 

  • Nukada Y, Ashikaga T, Sakaguchi H, Sono S, Mugita N, Hirota M, Miyazawa M, Ito Y, Sasa H, Nishiyama N (2011) Predictive performance for human skin sensitizing potential of the human cell line activation test (h-CLAT). Contact Dermatitis 65:343–353

    Article  CAS  PubMed  Google Scholar 

  • OECD (2002) Test No. 429: skin sensitisation: local lymph node assay 1–7

  • OECD (2012) The adverse outcome pathway for skin sensitisation initiated by covalent binding for proteins. Part 1. Scientific evidence OECD environment, health and safety publications series on testing and assessment

  • OECD (2015a) Test No. 442C: In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA)

  • OECD (2015b) Test No. 442D: In Vitro Skin Sensitisation. OECD Environ. Heal. Saf. Publ., pp 1–20

  • OECD (2016a) Test No. 442E: In Vitro Skin Sensitisation: Human Cell Line Activation Test (h-CLAT)

  • OECD (2016b) Guidance document on the reporting of defined approaches and individual information sources to be used within integrated approaches to testing and assessment (IATA) for skin sensitization, ENV/JM/MONO. OECD Publishing, Paris

    Google Scholar 

  • OECD (2017) Test No. 442E: in vitro skin sensitisation

  • Ouwehand K, Spiekstra SW, Reinders J, Scheper RJ, de Gruijl TD, Gibbs S (2010) Comparison of a novel CXCL12/CCL5 dependent migration assay with CXCL8 secretion and CD86 expression for distinguishing sensitizers from non-sensitizers using MUTZ-3 Langerhans cells. Toxicol In Vitro 24:578–585

    Article  CAS  PubMed  Google Scholar 

  • Python F, Goebel C, Aeby P (2007) Assessment of the U937 cell line for the detection of contact allergens. Toxicol Appl Pharmacol 220:113–124

    Article  CAS  PubMed  Google Scholar 

  • Python F, Goebel C, Aeby P (2009) Comparative DNA microarray analysis of human monocyte derived dendritic cells and MUTZ-3 cells exposed to the moderate skin sensitizer cinnamaldehyde. Toxicol Appl Pharmacol 239:273–283

    Article  CAS  PubMed  Google Scholar 

  • Raffalli C, Clouet E, Kuresepi S, Damiens M-H, Lepoittevin J-P, Pallardy M, Ferret P-J, Giménez-Arnau E, Kerdine-Römer S (2017) Fragrance allergens linalool and limonene allylic hydroperoxides in skin allergy: mechanisms of action focusing on transcription factor Nrf2. Toxicol Sci 1–10

  • Reich K, Westphal G, Konig IR, Mossner R, Kruger U, Ziegler A, Neumann C, Schnuch A (2003) Association of allergic contact dermatitis with a promoter polymorphism in the IL16 gene. J Allergy Clin Immunol 112:1191–1194

    Article  CAS  PubMed  Google Scholar 

  • Saito K, Takenouchi O, Nukada Y, Miyazawa M, Sakaguchi H (2017) An in vitro skin sensitization assay termed EpiSensA for broad sets of chemicals including lipophilic chemicals and pre/pro-haptens. Toxicol Vitr 40:11–25

    Article  CAS  Google Scholar 

  • Sakaguchi H, Ashikaga T, Miyazawa M, Yoshida Y, Ito Y, Yoneyama K, Hirota M, Itagaki H, Toyoda H, Suzuki H (2006) Development of an in vitro skin sensitization test using human cell lines; human Cell Line Activation Test (h-CLAT). II. an inter-laboratory study of the h-CLAT. Toxicol In Vitro 20:774–784

    Article  CAS  PubMed  Google Scholar 

  • Stepkowski TM, Kruszewski MK (2011) Molecular cross-talk between the NRF2/KEAP1 signaling pathway, autophagy, and apoptosis. Free Radic Biol Med 50:1186–1195

    Article  CAS  PubMed  Google Scholar 

  • Taub DD, Proost P, Murphy WJ, Anver M, Longo DL, van Damme J, Oppenheim JJ (1995) Monocyte chemotactic protein-1 (MCP-1), -2, and – 3 are chemotactic for human T lymphocytes. J Clin Invest 95:1370–1376

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Uguccioni M, D’Apuzzo M, Loetscher M, Dewald B, Baggiolini M (1995) Actions of the chemotactic cytokines MCP-1, MCP-2, MCP-3, RANTES, MIP-1 alpha and MIP-1 beta on human monocytes. Eur J Immunol 25:64–68

    Article  CAS  PubMed  Google Scholar 

  • Vandebriel RJ, Loveren H (2010) Non-animal sensitization testing: state-of-the-art. Crit Rev Toxicol 40:389–404

    Article  CAS  PubMed  Google Scholar 

  • Williams EH, Williams C, McLeod JD (2010) Identification of PDL-1 as a novel biomarker of sensitizer exposure in dendritic-like cells. Toxicol In Vitro 24:1727–1735

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Rami Bechara and Chloé Raffalli are contributed equally

Authors and Affiliations

  1. Safety Assessment Department, Pierre Fabre Dermo Cosmétique, Toulouse, France

    Elodie Clouet & Pierre-Jacques Ferret

  2. UMR996-Inflammation, Chemokines and Immunopathology, INSERM, Univ Paris-Sud, Université Paris-Saclay, 92296, Châtenay-Malabry, France

    Elodie Clouet, Rami Bechara, Chloé Raffalli, Marie-Hélène Damiens, Marc Pallardy & Saadia Kerdine-Römer

  3. ImmunoSearch, Grasse, France

    Hervé Groux

Authors
  1. Elodie Clouet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rami Bechara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chloé Raffalli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marie-Hélène Damiens
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hervé Groux
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marc Pallardy
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pierre-Jacques Ferret
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Saadia Kerdine-Römer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Saadia Kerdine-Römer.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest and also no sponsorship.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 40 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Clouet, E., Bechara, R., Raffalli, C. et al. The THP-1 cell toolbox: a new concept integrating the key events of skin sensitization. Arch Toxicol 93, 941–951 (2019). https://doi.org/10.1007/s00204-019-02416-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-019-02416-7

Keywords

Search

Navigation