The Human T Cell Priming Assay (hTCPA)

  • Philipp R. Esser
  • Stefan F. Martin


The induction of allergic contact dermatitis by chemicals depends on their ability to activate a chemical-specific T cell response. This key event that concludes the skin sensitization process is monitored in an in vitro assay, the human T cell priming assay (hTCPA). Naive human T cells are incubated with autologous dendritic cells pulsed with the chemical of interest. After restimulation with the same chemical, the induction of an antigen-specific T cell response can be assessed by the detection of T cell proliferation, cytokine production or surface marker expression. The hTCPA still needs optimization but may nevertheless be a valuable in vitro assay within an integrated testing strategy aiming at replacement of animal testing for contact allergen identification.


Chemical Contact allergen T cell Skin In vitro assay Animal testing 


  1. 1.
    Peiser M, Tralau T, Heidler J, Api AM, Arts JHE, Basketter DA, English J, Diepgen TL, Fuhlbrigge RC, Gaspari AA, et al. Allergic contact dermatitis: epidemiology, molecular mechanisms, in vitro methods and regulatory aspects. Cell Mol Life Sci. 2011;69:763–81.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Kimber I, Dearman RJ, Basketter DA, Ryan CA, Gerberick GF. The local lymph node assay: past, present and future. Contact Dermatol. 2002;47:315–28.CrossRefGoogle Scholar
  3. 3.
    Kehren J, Desvignes C, Krasteva M, Ducluzeau MT, Assossou O, Horand F, Hahne M, Kägi D, Kaiserlian D, Nicolas JF. Cytotoxicity is mandatory for CD8(+) T cell-mediated contact hypersensitivity. J Exp Med. 1999;189:779–86.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Martin S, Lappin MB, Kohler J, Delattre V, Leicht C, Preckel T, Simon JC, Weltzien HU. Peptide immunization indicates that CD8+ T cells are the dominant effector cells in trinitrophenyl-specific contact hypersensitivity. J Invest Dermatol. 2000;115:260–6.CrossRefPubMedGoogle Scholar
  5. 5.
    Martin SF, et al. T-cell recognition of chemicals, protein allergens and drugs: towards the development of in vitro assays. Cell Mol Life Sci. 2010;67:4171–84.CrossRefPubMedGoogle Scholar
  6. 6.
    Vocanson M, Hennino A, Rozieres A, Cluzel-Tailhardat M, Poyet G, Valeyrie M, Benetiere J, Tedone R, Kaiserlian D, Nicolas J-F. Skin exposure to weak and moderate contact allergens induces IFN[gamma] production by lymph node cells of CD4+ T-cell-depleted mice. J Invest Dermatol. 2008;129:1185–91.CrossRefPubMedGoogle Scholar
  7. 7.
    Cavani A, Pennino D, Eyerich K. Th17 and Th22 in skin allergy. In: Ring J, Darsow U, Behrendt H, editors. Chemical immunology and allergy. KARGER: Basel; 2012. p. 39–44.Google Scholar
  8. 8.
    Reisinger K, Hoffmann S, Alépée N, Ashikaga T, Barroso J, Elcombe C, Gellatly N, Galbiati V, Gibbs S, Groux H, et al. Systematic evaluation of non-animal test methods for skin sensitisation safety assessment. Toxicol In Vitro. 2015;29:259–70.CrossRefPubMedGoogle Scholar
  9. 9.
    Van der Veen JW, Rorije E, Emter R, Natsch A, van Loveren H, Ezendam J. Evaluating the performance of integrated approaches for hazard identification of skin sensitizing chemicals. Regul Toxicol Pharmacol. 2014;69:371–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Robinson MK. Optimization of an in vitro lymphocyte blastogenesis assay for predictive assessment of immunologic responsiveness to contact sensitizers. J Invest Dermatol. 1989;92:860–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Mattarollo SR, Yong M, Gosmann C, Choyce A, Chan D, Leggatt GR, Frazer IH. NKT cells inhibit antigen-specific effector CD8 T cell induction to skin viral proteins. J Immunol. 2011;187:1601–8.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Goubier A, Vocanson M, Macari C, Poyet G, Herbelin A, Nicolas J-F, Dubois B, Kaiserlian D. Invariant NKT cells suppress CD8+ T-cell-mediated allergic contact dermatitis independently of regulatory CD4+ T cells. J Invest Dermatol. 2013;133:980–7.CrossRefPubMedGoogle Scholar
  13. 13.
    Vocanson M, Achachi A, Mutez V, Cluzel-Tailhardat M, Varlet BL, Rozières A, Fournier P, Nicolas J-F. Human T cell priming assay: depletion of peripheral blood lymphocytes in CD25(+) cells improves the in vitro detection of weak allergen-specific T cells. EXS. 2014;104:89–100.PubMedGoogle Scholar
  14. 14.
    Dietz L, Esser PR, Schmucker SS, Goette I, Richter A, Schnölzer M, Martin SF, Thierse H-J. Tracking human contact allergens: from mass spectrometric identification of peptide-bound reactive small chemicals to chemical-specific naive human T-cell priming. Toxicol Sci. 2010;117:336–47.CrossRefPubMedGoogle Scholar
  15. 15.
    Richter A, Schmucker SS, Esser PR, Traska V, Weber V, Dietz L, Thierse H-J, Pennino D, Cavani A, Martin SF. Human T cell priming assay (hTCPA) for the identification of contact allergens based on naive T cells and DC–IFN-γ and TNF-α readout. Toxicol In Vitro. 2013;27:1180–5.CrossRefPubMedGoogle Scholar
  16. 16.
    Bonefeld CM, Nielsen MM, Rubin IMC, Vennegaard MT, Dabelsteen S, Gimenéz-Arnau E, Lepoittevin J-P, Geisler C, Johansen JD. Enhanced sensitization and elicitation responses caused by mixtures of common fragrance allergens. Contact Dermatitis. 2011;65:336–42.CrossRefPubMedGoogle Scholar
  17. 17.
    Johansen JD, Skov L, Volund A, Andersen K, Menné TJ. Allergens in combination have a synergistic effect on the elicitation response: a study of fragrance-sensitized individuals. Br J Dermatol. 1998;139:264–70.CrossRefPubMedGoogle Scholar
  18. 18.
    De Jong WH, Tentij M, Spiekstra SW, Vandebriel RJ, Van Loveren H. Determination of the sensitising activity of the rubber contact sensitisers TMTD, ZDMC, MBT and DEA in a modified local lymph node assay and the effect of sodium dodecyl sulfate pretreatment on local lymph node responses. Toxicology. 2002;176:123–34.CrossRefPubMedGoogle Scholar
  19. 19.
    Vocanson M, Rozieres A, Hennino A, Poyet G, Gaillard V, Renaudineau S, Achachi A, Benetiere J, Kaiserlian D, Dubois B, et al. Inducible costimulator (ICOS) is a marker for highly suppressive antigen-specific T cells sharing features of TH17/TH1 and regulatory T cells. J Allergy Clin Immunol. 2010;126:280–289.e7.CrossRefPubMedGoogle Scholar
  20. 20.
    Faulkner L, Martinsson K, Santoyo-Castelazo A, Cederbrant K, Schuppe-Koistinen I, Powell H, Tugwood J, Naisbitt DJ, Park BK. The development of in vitro culture methods to characterize primary T-cell responses to drugs. Toxicol Sci. 2012;127:150–8.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  1. 1.Allergy Research Group, Department of Dermatology, Medical Center-University of FreiburgFaculty of Medicine, University of FreiburgFreiburgGermany

Personalised recommendations