Identification of Skin Allergens by In Vivo Assay

  • David A. BasketterEmail author
Reference work entry


In vivo assays for skin sensitization are valuable assets, but must not be regarded as 100% accurate.

Guinea pig methods have been superseded by the murine local lymph node assay (LLNA) which is now in the process of being replaced by nonanimal alternatives.

Regulatory classification as a skin sensitizing chemical means that the substance is regarded as significantly active.

A substance which is not classified as a skin sensitizer may still possess a limited potential to cause this effect.

Potency categories are new to regulatory sensitization, but the LLNA EC3 value provides a useful guide to the relative potency of a skin sensitizer.


Skin sensitization Local lymph node assay Regulatory classification Potency categorization 


  1. Api AM, Basketter DA, Cadby PA, Cano M-F, Ellis G, Gerberick GF, Griem P, McNamee PM, Ryan CA, Safford B (2008) Dermal sensitization quantitative risk assessment (QRA) for fragrance ingredients. Regul Toxicol Pharmacol 52:3–23CrossRefGoogle Scholar
  2. Api AM, Parakhia R, O’Brien D, Basketter DA (2017) Fragrances categorised according to their relative human skin sensitization potency. Dermatitis, 28:299–307CrossRefGoogle Scholar
  3. Basketter DA (2008) Skin immunology and sensitisation. In: Chilcott RP, Price S (eds) Principles and practice of skin toxicology. Wiley, Chichester, pp 149–168CrossRefGoogle Scholar
  4. Basketter DA (2010) Methyldibromo glutaronitrile, skin sensitisation and quantitative risk assessment. Cutan Ocul Toxicol 29:4–9CrossRefGoogle Scholar
  5. Basketter DA, Gerberick GF (1996) Interlaboratory evaluation of the Buehler test. Contact Dermatitis 35:146–151CrossRefGoogle Scholar
  6. Basketter DA, Kimber I (2010) Skin sensitization, false positives and false negatives: experience with guinea pig assays. J Appl Toxicol 30(5):381–386PubMedGoogle Scholar
  7. Basketter DA, Gerberick GF, Kimber I (1998) Strategies for identifying false positive responses in predictive sensitization tests. Food Chem Toxicol 36:327–333CrossRefGoogle Scholar
  8. Basketter DA, Lea L, Cooper K, Dickens A, Briggs D, Pate I, Dearman RJ, Kimber I (1999) A comparison of statistical approaches to derivation of EC3 values from local lymph node assay dose responses. J Appl Toxicol 19:261–266CrossRefGoogle Scholar
  9. Basketter DA, Andersen KE, Lidén C, van Loveren H, Boman A, Kimber I, Alanko K, Berggren E (2005) Evaluation of the skin sensitising potency of chemicals using existing methods and considerations of relevance for elicitation. Contact Dermatitis 52:39–43CrossRefGoogle Scholar
  10. Basketter DA, McFadden J, Evans P, Andersen KE, Jowsey I (2006) Identification and classification of skin sensitizers: identifying false positives and false negatives. Contact Dermatitis 55:268–273CrossRefGoogle Scholar
  11. Basketter DA, Gerberick GF, Kimber I (2007) The local lymph node assay EC3 value: status of validation. Contact Dermatitis 57:70–75CrossRefGoogle Scholar
  12. Basketter DA, Ball N, Cagen S, Carrillo J-C, Certa H, Eigler D, Esch H, Graham C, Haux D, Kreiling R, Mehling A (2009a) Application of a weight of evidence approach to analysing discordant sensitization datasets: implication for REACH. Regul Toxicol Pharmacol 55:90–96CrossRefGoogle Scholar
  13. Basketter DA, McFadden JF, Gerberick GF, Cockshott A, Kimber I (2009b) Nothing is perfect, not even the local lymph node assay. A commentary and the implications for REACH. Contact Dermatitis 60:65–69CrossRefGoogle Scholar
  14. Basketter DA, Alepee N, Ashikaga T, Barroso J, Gilmour N, Goebel C, Hibatallah J, Hoffmann S, Kern P, Martinozzi-Teissier S, Maxwell G, Millet M, Reisinger K, Sakaguchi H, Schepky A, Tialhardat M, Templier M (2014) Categorisation of chemicals according to their relative human skin sensitizing potency. Dermatitis, 25: 11–21CrossRefGoogle Scholar
  15. Basketter DA, White IR, McFadden JP, Kimber I (2015) Skin sensitization: integration of clinical data into hazard identification and risk assessment. Hum Exp Toxicol 34:1222–1230CrossRefGoogle Scholar
  16. Buehler EV (1965) Delayed contact hypersensitivity in the guinea pig. Arch Dermatol 91:171–177CrossRefGoogle Scholar
  17. Casati S, Aeby P, Kimber I, Maxwell G, Ovigne JM, Roggen E, Rovida C, Tosti L, Basketter DA (2009) Selection of chemicals for the development and evaluation of in vitro methods for skin sensitisation testing. Altern Lab Anim 37:305–312CrossRefGoogle Scholar
  18. Commission of the European Communities Regulation (2006) (EC) No 1907/2006 of the European parliament and of the council of 18 December 2006 concerning the registration, evaluation, authorisation and restriction of chemicals (REACH), establishing a European chemicals agency, amending Directive 1999/45/EC and repealing council regulation (EEC) No 793/93 and commission regulation (EC) No 1488/94 as well as council directive 76/769/EEC and commission directives 91/155/EEC, 93/67/EEC,93/105/EC and 2000/21/EC. Off J Eur Union L 396/1 of 30 Dec 2006.
  19. Cronin MTD, Basketter DA (1994) Multivariate QSAR analysis of a skin sensitization database. SAR QSAR Environ Res 2:159–179CrossRefGoogle Scholar
  20. Dean JH, Twerdok LE, Tice RR, Sailstad DM, Hattan DG, Stokes WS (2001) ICCVAM evaluation of the murine local lymph node assay. II. Conclusions and recommendations of an independent scientific peer review panel. Regul Toxicol Pharmacol 34:258–273CrossRefGoogle Scholar
  21. ECHA (2015) Chapter R.7a: endpoint specific guidance. In: Guidance on information requirements and chemical safety assessment. European Chemicals Agency, Helsinki, pp 256–288Google Scholar
  22. ECHA (2017) Guidance on Information Requirements and Chemical Safety Assessment. Chapter R7a: Endpoint specific guidance. Version 6.0, July 2017. Last accessed May 29th 2018.
  23. Felter SP, Robinson MK, Basketter DA, Gerberick GF (2002) A review of the scientific basis for default uncertainty factors for use in quantitative risk assessment of the induction of allergic contact dermatitis. Contact Dermatitis 47:257–266CrossRefGoogle Scholar
  24. Felter SP, Ryan CA, Basketter DA, Gerberick GF (2003) Application of the risk assessment paradigm to the induction of allergic contact dermatitis. Regul Toxicol Pharmacol 37:1–10CrossRefGoogle Scholar
  25. Frankild S, Basketter DA, Andersen KE (1996) The value and limitations of rechallenge in the guinea pig maximisation test. Contact Dermatitis 35:135–140CrossRefGoogle Scholar
  26. Gerberick GF, Ryan CA, Kimber I, Dearman RJ, Lea LJ, Basketter DA (2000) Local lymph node assay: validation assessment for regulatory purposes. Am J Contact Dermat 11:3–18CrossRefGoogle Scholar
  27. Gerberick GF, Ryan CA, Kern PS, Schlatter H, Dearman RJ, Kimber I, Patlewicz G, Basketter DA (2005) Compilation of historical local lymph node assay data for the evaluation of skin sensitization alternatives. Dermatitis 16:157–202PubMedGoogle Scholar
  28. Kern PS, Gerberick GF, Ryan CA, Kimber I, Aptula A, Basketter DA (2010) Historical local lymph node data for the evaluation of skin sensitization alternatives: a second compilation. Dermatitis 21:8–32CrossRefGoogle Scholar
  29. Kimber I, Basketter DA (1992) The murine local lymph node assay; collaborative studies and new directions: A commentary. Food Chem Toxicol 30, 165–169.Google Scholar
  30. Kligman AM, Basketter DA (1995) A critical commentary and updating of the guinea pig maximisation test. Contact Dermatitis 32:129–134CrossRefGoogle Scholar
  31. Kreiling R, Hollnagel HM, Hareng L, Eigler D, Lee MS, Griem P, Dreesen B, Kleber M, Albrecht A, Garcia C, Wendel A (2008) Comparison of the skin sensitizing potential of unsaturated compounds and assessed by the murine local lymph node assay (LLNA) and the guinea pig maximization test (GPMT). Food Chem Toxicol 46:1896–1904CrossRefGoogle Scholar
  32. Magnusson B, Kligman AM (1970) Allergic contact dermatitis in the guinea pig: identification of contact allergens. Charles C. Thomas, Springfield, 141ppGoogle Scholar
  33. McGarry HF (2007) The murine local lymph node assay: regulatory and potency considerations under REACH. Toxicology 238:71–89CrossRefGoogle Scholar
  34. NICEATM (2010) LLNA Database. Available at Last accessed May 29th 2018
  35. OECD (Organisation for Economic Cooperation and Development) (1992) Guidelines for testing of chemicals no. 406. Skin sensitization. Organisation for Economic Cooperation and Development, ParisGoogle Scholar
  36. OECD (Organisation for Economic Cooperation and Development) (2010a) Test guideline 429: the local lymph node assay. OECD, ParisGoogle Scholar
  37. OECD (Organisation for Economic Cooperation and Development) (2010b) Test guideline 442A and 442B: the local lymph node assay DA and BrdU ELISA. OECD, ParisGoogle Scholar
  38. United Nations (2009) Globally harmonized system of classification and labelling of chemicals (GHS). Part 3: health hazards. United Nations, New York. Scholar
  39. van Loveren H, Cockshott A, Gebel T, Gundert-Remy U, de Jong WH, Matheson J, McGarry H, Musset L, Selgrade MK, Vickers C (2008) Skin sensitization in chemical risk assessment: report of a WHO/IPCS international workshop focusing on dose–response assessment. Regul Toxicol Pharmacol 50(2):155–199CrossRefGoogle Scholar
  40. Wahlberg JE, Boman A (1985) Guinea pig maximization test. Curr Probl Dermatol 14:59–106CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.DABMEB Consultancy Ltd.SharnbrookUK

Personalised recommendations