In Vitro Delayed-Type Sensitivity Testing

  • Detlef BeckerEmail author
Reference work entry


Although patch testing is regarded to be the most standardized and reliable method for diagnosis of delayed-type hypersensitivity to contact allergens, its substitution by in vitro techniques would be advantageous for several reasons.

The lymphocyte transformation test (LTT) measures the allergen-specific proliferation or activation of T cells in vitro and has been explored as potential alternative for patch testing for decades. Beside numerous attempts to increase its sensitivity and specificity, the major drawback of a cell culture-based method is the poor solubility of many contact sensitizers and the need for chemical modification of prohaptens.

The LTT is used with regularity only for diagnosis of hypersensitivity to beryllium and adverse reactions to drugs. Its usefulness in occupational dermatology is limited and restricted to special situations as a supplement for patch testing with large panels of well-established sensitizers.


Patch testing Lymphocyte transformation test Delayed-type hypersensitivity Beryllium Drug hypersensitivity In vitro testing MELISA Nickel Mercury T cells ELISpot Cytokines Cell proliferation Sensitivity Specificity 





Lymphocyte transformation test


Peripheral blood mononuclear cells


  1. Bordignon V, Palamara F, Cordiali-Fei P, Vento A, Aiello A, Picardo M, Ensoli F, Cristaudo A (2008) Nickel, palladium and rhodium induced IFN-gamma and IL-10 production as assessed by in vitro ELISpot-analysis in contact dermatitis patients. BMC Immunol 9:19. Scholar
  2. Bordignon V, Palamara F, Altomonte G, Sperduti I, Pietravalle M, Cavallotti C, Cordiali-Fei P, Fuggetta MP, Cristaudo A, Ensoli F (2015) A laboratory test based on determination of cytokine profiles: a promising assay to identify exposition to contact allergens and predict the clinical outcome in occupational allergic contact dermatitis. BMC Immunol 16:4. Scholar
  3. Brand CU, Schmidli J, Ballmer-Weber B, Hunziker T (1995) Lymphocyte stimulation test, a possible alternative for verifying chloroacetophenone sensitization. Hautarzt 46:702–704CrossRefGoogle Scholar
  4. Byers VS, Castagnoli N Jr, Epstein WL (1979) In vitro studies of poison oak immunity. II. Effect of urushiol analogues on the human in vitro response. J Clin Invest 64:1449–1456CrossRefGoogle Scholar
  5. Cederbrant K, Hultman P, Marcusson JA, Tibbling L (1997) In vitro lymphocyte proliferation as compared to patch test using gold, palladium and nickel. Int Arch Allergy Immunol 112:212–217CrossRefGoogle Scholar
  6. Cederbrant K, Gunnarsson LG, Hultman P, Norda R, Tibbling-Grahn L (1999) In vitro lymphoproliferative assays with HgCl2 cannot identify patients with systemic symptoms attributed to dental amalgam. J Dent Res 78:1450–1458CrossRefGoogle Scholar
  7. Chipinda I, Hettick JM, Siegel PD (2011) Haptenation: chemical reactivity and protein binding. J Allergy (Cairo) 2011:839682. Scholar
  8. Doña I, Torres MJ, Montañez MI, Fernández TD (2017) In vitro diagnostic testing for antibiotic allergy. Allergy Asthma Immunol Res 9:288–298. Scholar
  9. Farris GM, Newman LS, Frome EL, Shou Y, Barker E, Habbersett RC, Maier L, Smith HN, Marrone BL (2000) Detection of beryllium sensitivity using a flow cytometric lymphocyte proliferation test: the Immuno-Be-LPT. Toxicology 143:125–140CrossRefGoogle Scholar
  10. Fireman E, Shai AB, Alcalay Y, Ophir N, Kivity S, Stejskal V (2016) Identification of metal sensitization in sarcoid-like metal-exposed patients by the MELISA® lymphocyte proliferation test – a pilot study. J Occup Med Toxicol 11:18. Scholar
  11. Ghatan PH, Marcusson-Ståhl M, Matura M, Björkheden C, Lundborg P, Cederbrant K (2014) Sensitization to omeprazole in the occupational setting. Contact Dermatitis 71:371–375. Scholar
  12. Jakobson E, Masjedi K, Ahlborg N, Lundeberg L, Karlberg AT, Scheynius A (2002) Cytokine production in nickel-sensitized individuals analysed with enzyme-linked immunospot assay: possible implication for diagnosis. Br J Dermatol 147:442–449CrossRefGoogle Scholar
  13. Kano Y, Hirahara K, Mitsuyama Y, Takahashi R, Shiohara T (2007) Utility of the lymphocyte transformation test in the diagnosis of drug sensitivity: dependence on its timing and the type of drug eruption. Allergy 62:1439–1444CrossRefGoogle Scholar
  14. Kneilling M, Caroli U, Grimmel C, Fischer J, Eichner M, Wieder T, Maier FC, Röcken M, Biedermann T (2010) Para-phenylenediamine-specific lymphocyte activation test: a sensitive in vitro assay to detect para-phenylenediamine sensitization in patients with severe allergic reactions. Exp Dermatol 19:435–441CrossRefGoogle Scholar
  15. Krasteva M, Nicolas JF, Chabeau G, Garrigue JL, Bour H, Thivolet J, Schmitt D (1993) Dissociation of allergenic and immunogenic functions in contact sensitivity to para-phenylenediamine. Int Arch Allergy Immunol 102:200–204CrossRefGoogle Scholar
  16. Lindemann M, Böhmer J, Zabel M, Grosse-Wilde H (2003) ELISpot: a new tool for the detection of nickel sensitization. Clin Exp Allergy 33:992–998CrossRefGoogle Scholar
  17. Lisby S, Hansen LH, Menn T, Baadsgaard O (1999) Nickel-induced proliferation of both memory and naive T cells in patch test-negative individuals. Clin Exp Immunol 117:217–222CrossRefGoogle Scholar
  18. Martins LE, Duarte AJ, Aoki V, Nunes RS, Ogusuku S, Reis VM (2008) Lymphocyte proliferation testing in chromium allergic contact dermatitis. Clin Exp Dermatol 33:472–477CrossRefGoogle Scholar
  19. Masjedi K, Ahlborg N, Gruvberger B, Bruze M, Karlberg AT (2003) Methylisothiazolinones elicit increased production of both T helper (Th)1- and Th2-like cytokines by peripheral blood mononuclear cells from contact allergic individuals. Br J Dermatol 149:1172–1182CrossRefGoogle Scholar
  20. Merk HF (2005) Diagnosis of drug hypersensitivity: lymphocyte transformation test and cytokines. Toxicology 209:217–220CrossRefGoogle Scholar
  21. Middleton D, Kowalski P (2010) Advances in identifying beryllium sensitization and disease. Int J Environ Res Public Health 7:115–124CrossRefGoogle Scholar
  22. Milovanova TN, Popma SH, Cherian S, Moore JS, Rossman MD (2004) Flow cytometric test for beryllium sensitivity. Cytometry B Clin Cytom 60:23–30CrossRefGoogle Scholar
  23. Moed H, von Blomberg M, Bruynzeel DP, Scheper R, Gibbs S, Rustemeyer T (2005) Improved detection of allergen-specific T-cell responses in allergic contact dermatitis through the addition of ‘cytokine cocktails’. Exp Dermatol 14:634–640CrossRefGoogle Scholar
  24. Mroz MM, Maier LA, Strand M, Silviera L, Newman LS (2009) Beryllium lymphocyte proliferation test surveillance identifies clinically significant beryllium disease. Am J Ind Med 52:762–773CrossRefGoogle Scholar
  25. Pichler WJ, Tilch J (2004) The lymphocyte transformation test in the diagnosis of drug hypersensitivity. Allergy 59:809–820CrossRefGoogle Scholar
  26. Popple A, Williams J, Maxwell G, Gellatly N, Dearman RJ, Kimber I (2016) T lymphocyte dynamics in methylisothiazolinone-allergic patients. Contact Dermatitis 75:1–13. Scholar
  27. Räsänen L, Tuomi ML (1992) Diagnostic value of the lymphocyte proliferation test in nickel contact allergy and provocation in occupational coin dermatitis. Contact Dermatitis 27:250–254CrossRefGoogle Scholar
  28. Räsänen L, Sainio H, Lehto M, Reunala T (1991) Lymphocyte proliferation test as a diagnostic aid in chromium contact sensitivity. Contact Dermatitis 25:25–29CrossRefGoogle Scholar
  29. Riviera AP, Pezzini A, Zanoni G, Rocca M, Pagani M, Tridente G (1995) Immunological reactivity in ranitidine factory workers. Int Arch Occup Environ Health 66:407–411CrossRefGoogle Scholar
  30. Spiewak R, Moed H, von Blomberg BM, Bruynzeel DP, Scheper RJ, Gibbs S, Rustemeyer T (2007) Allergic contact dermatitis to nickel: modified in vitro test protocols for better detection of allergen-specific response. Contact Dermatitis 56:63–69CrossRefGoogle Scholar
  31. Ständer S, Oppel E, Thomas P, Summer B (2017) Evaluation of lymphocyte transformation tests as compared with patch tests in nickel allergy diagnosis. Contact Dermatitis 76:228–234. Scholar
  32. Stejskal VD, Olin RG, Forsbeck M (1986) The lymphocyte transformation test for diagnosis of drug-induced occupational allergy. J Allergy Clin Immunol 77:411–426CrossRefGoogle Scholar
  33. Stejskal VD, Forsbeck M, Nilsson R (1990) Lymphocyte transformation test for diagnosis of isothiazolinone allergy in man. J Invest Dermatol 94:798–802CrossRefGoogle Scholar
  34. Stejskal VD, Cederbrant K, Lindvall A, Forsbeck M (1994) MELISA-an in vitro tool for the study of metal allergy. Toxicol In Vitro 8:991–1000CrossRefGoogle Scholar
  35. Valentine-Thon E, Müller K, Guzzi G, Kreisel S, Ohnsorge P, Sandkamp M (2006) LTT-MELISA is clinically relevant for detecting and monitoring metal sensitivity. Neuro Endocrinol Lett 27(Suppl 1):17–24PubMedGoogle Scholar
  36. Yawalkar N, Helbling A, Pichler CE, Zala L, Pichler WJ (1999) T cell involvement in persulfate triggered occupational contact dermatitis and asthma. Ann Allergy Asthma Immunol 82:401–404CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.Department of DermatologyUniversity Medical CenterMainzGermany

Personalised recommendations