A triangular approach for the validation of new approach methods for skin sensitization

Main Article Content

Andreas Natsch , Robert Landsiedel, Susanne N. Kolle
[show affiliations]


The availability of reference data is a key requirement for the development of new approach methods (NAM), i.e., in vitro, in chemico and in silico methods and integrated approaches, like defined approaches (DA), which combine these data sources. Reference data are of even greater importance for regulatory acceptance. In contrast to most other adverse effects, human skin sensitization data on many chemicals are available, next to data from animal studies, such as the local lymph node assay (LLNA). Skin sensitization NAM data can therefore be compared to different reference datasets. Recent publications and validation at the OECD focused on human and LLNA reference data. The “2 out of 3” DA (2o3 DA) is the first DA for skin sensitization solely based on experimental data from validated tests and was recently adopted as an OECD test guideline. Here we review the predictivity of the 2o3 DA on multiple human and LLNA reference datasets. Concomitantly, we compare the predictivity of the LLNA for human data within the same datasets. Comparing predictivity of methods not only bilaterally (NAM or DA vs. animal method) but including human data in a triangle “NAM data – animal data – human data” offers a comprehensive assessment of the NAM’s and DA’s predictivity. In all these assessments, the 2o3 DA was superior to the LLNA in predicting human skin sensitization hazard. This highlights the importance of a holistic view of reference data instead of limiting validation of NAMs and DAs to data from a single animal test only.

Article Details

How to Cite
Natsch, A., Landsiedel, R. and Kolle, S. N. (2021) “A triangular approach for the validation of new approach methods for skin sensitization”, ALTEX - Alternatives to animal experimentation, 38(4), pp. 669–677. doi: 10.14573/altex.2105111.

Basketter, D. A., Alepee, N., Ashikaga, T., Barroso, J. et al. (2014). Categorization of chemicals according to their rela-tive human skin sensitizing potency. Dermatitis 25, 11-21. doi:10.1097/der.0000000000000003

Basketter, D. A., Lea, L. J., Cooper, K., Stocks, J. et al. (1999). Threshold for classification as a skin sensitizer in the local lymph node assay: A statistical evaluation. Food and Chemical Toxicology 37, 1167-1174. doi:10.1016/s0278-6915(99)00112-x

Bauch, C., Kolle, S. N., Ramirez, T., Eltze, T. et al. (2012). Putting the parts together: Combining in vitro methods to test for skin sensitizing potentials. Regul Toxicol Pharmacol 63, 489-504. doi:10.1016/j.yrtph.2012.05.013; Corri-gendum: doi:10.1016/j.yrtph.2012.08.014

Casati, S., Aeby, P., Kimber, I., Maxwell, G. et al. (2009). Selection of chemicals for the development and evaluation of in vitro methods for skin sensitisation testing. Altern Lab Anim 37, 305-12. doi:10.1177/026119290903700313

Dimitrov, S. D., Low, L. K., Patlewicz, G. Y., Kern, P. S. et al. (2005). Skin sensitization: Modeling based on skin me-tabolism simulation and formation of protein conjugates. International Journal of Toxicology 24, 189-204. doi:10.1080/10915810591000631

EC - European Commission (2009). Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products. Official Journal of the European Union https://eur-lex.europa.eu/eli/reg/2009/1223/oj/eng.

EC (2013). Full EU ban on animal testing for cosmetics enters into force. https://ec.europa.eu/commission/presscorner/detail/en/IP_13_210.

Emter, R., Ellis, G. and Natsch, A. (2010). Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro. Toxicol. Appl. Pharmacol. 245, 281-290. doi:10.1016/j.taap.2010.03.009

Ezendam, J., Braakhuis, H. M. and Vandebriel, R. J. (2016). State of the art in non-animal approaches for skin sensitiza-tion testing: from individual test methods towards testing strategies. Arch Toxicol. doi:10.1007/s00204-016-1842-4

Gabbert, S., Mathea, M., Kolle, S. N. and Landsiedel, R. (2020). Accounting for Precision Uncertainty of Toxicity Test-ing: Methods to Define Borderline Ranges and Implications for Hazard Assessment of Chemicals. Risk Anal. doi:10.1111/risa.13648

Gray, L. E., Jr., Furr, J. R., Lambright, C. S., Evans, N. et al. (2020). Quantification of the uncertainties in extrapolating from in vitro androgen receptor antagonism to in vivo hershberger assay endpoints and adverse reproductive development in male rats. Toxicological Sciences 176, 297-311. doi:10.1093/toxsci/kfaa067

Haneke, K. E., Tice, R. R., Carson, B. L., Margolin, B. H. et al. (2001). ICCVAM evaluation of the murine local lymph node assay. Data analyses completed by the National Toxicology Program Interagency Center for the Evalua-tion of Alternative Toxicological Methods. Regul Toxicol Pharmacol 34, 274-86. doi:10.1006/rtph.2001.1498

Hoffmann, S., Kleinstreuer, N., Alepee, N., Allen, D. et al. (2018). Non-animal methods to predict skin sensitization (I): the Cosmetics Europe database(). Crit. Rev. Toxicol. in press, 1-15. doi:10.1080/10408444.2018.1429385

ICCVAM (1999). The Murine Local Lymph Node Assay: A Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals/Compounds. The Results of an Independent Peer Review Evaluation Coordinated by the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the National Toxicology Program Center for the Evaluation of Alternative Toxicological Methods (NICEATM). NIH Publi-cation No. 99-4494. doi:https://ntp.niehs.nih.gov/iccvam/docs/immunotox_docs/llna/llnarep.pdf

ICCVAM (2009). Recommended Performance Standards: Murine Local Lymph Node Assay ICCVAM report NIH Publi-cation No. 09-7357

Jaworska, J., Dancik, Y., Kern, P., Gerberick, F. et al. (2013). Bayesian integrated testing strategy to assess skin sensiti-zation potency: From theory to practice. J. Appl. Toxicol. 33, 1353-1364. doi:10.1002/jat.2869

Johansson, H., Albrekt, A. S., Borrebaeck, C. A. and Lindstedt, M. (2013). The GARD assay for assessment of chemical skin sensitizers. Toxicol In Vitro 27, 1163-9. doi:10.1016/j.tiv.2012.05.019

Jowsey, I. R., Basketter, D. A., Westmoreland, C. and Kimber, I. (2006). A future approach to measuring relative skin sensitising potency: A proposal. Journal of Applied Toxicology 26, 341-350. doi:10.1002/jat.1146

Kleinstreuer, N. C., Hoffmann, S., Alepee, N., Allen, D. et al. (2018). Non-animal methods to predict skin sensitization (II): an assessment of defined approaches (*). Crit Rev Toxicol, 1-16. doi:10.1080/10408444.2018.1429386

Kolle, S. N., Hill, E., Raabe, H., Landsiedel, R. et al. (2019). Regarding the references for reference chemicals of alter-native methods. Toxicology in Vitro 57, 48-53. doi:10.1016/j.tiv.2019.02.007

Kolle, S. N., Landsiedel, R. and Natsch, A. (2020). Replacing the refinement for skin sensitization testing: Considera-tions to the implementation of adverse outcome pathway (AOP)-based defined approaches (DA) in OECD guidelines. Regul Toxicol Pharmacol 115, 104713. doi:10.1016/j.yrtph.2020.104713

Kolle, S. N., Mathea, M., Natsch A., and Landsiedel R. (in press). Assessing Experimental Uncertainty in Defined Approaches: Borderline Ranges for In Chemico and In Vitro Skin Sensitization Methods Determined from Ring Trial Data. Appl In Vitro Toxicol. doi:10.1089/aivt.2021.0003

Lacroix, G., Koch, W., Ritter, D., Gutleb, A. C. et al. (2018). Air-Liquid Interface In Vitro Models for Respiratory Toxi-cology Research: Consensus Workshop and Recommendations. Appl In Vitro Toxicol 4, 91-106. doi:10.1089/aivt.2017.0034

Leontaridou, M., Urbisch, D., Kolle, S. N., Ott, K. et al. (2017). The borderline range of toxicological methods: Quantifi-cation and implications for evaluating precision. ALTEX 34, 525-538. doi:10.14573/altex.1606271

Natsch, A., Ryan, C. A., Foertsch, L., Emter, R. et al. (2013). A dataset on 145 chemicals tested in alternative assays for skin sensitization undergoing prevalidation. J Appl Toxicol 33, 1337-52. doi:10.1002/jat.2868

Nukada, Y., Miyazawa, M., Kazutoshi, S., Sakaguchi, H. et al. (2013). Data integration of non-animal tests for the de-velopment of a test battery to predict the skin sensitizing potential and potency of chemicals. Toxicology in Vitro 27, 609-618. doi:10.1016/j.tiv.2012.11.006

OECD (2018a). In vitro skin sensitisation assays addressing the key event on activation of dendritic cells on the adverse outcome pathway for skin sensitisation. OECD testing guidelines 442e. doi:10.1787/9789264264359-en

OECD (2018b). Test No. 442D: In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method. OECD Guidelines for the Testing of Chemicals No. 442D. doi:10.1787/9789264229822-en

OECD (2020). In Chemico Skin Sensitisation Assays addressing the Adverse Outcome Pathway, key event on covalent binding to proteins. OECD testing guidelines 442c. doi:10.1787/9789264229709-en

OECD (2021a). Guideline No. 497: Defined Approaches on Skin Sensitisation. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris https://www.oecd-ilibrary.org/environment/guideline-no-497-defined-approaches-on-skin-sensitisation_b92879a4-en. doi:10.1177/026119290703500311

OECD (2021b). Series on Testing and Assessment No. 336: Supporting document to the Guideline (GL) on Defined Approaches (DAs) for Skin Sensitisation- Annex 2. Organisation for Economic Cooperation and Development, Paris.

OECD (2021c). Series on Testing and Assessment No. 336: Supporting document to the Guideline (GL) on Defined Approaches (DAs) for Skin Sensitisation- Annex 3. Organisation for Economic Cooperation and Development, Paris.

OECD (2021d). Series on Testing and Assessment No. 336: Supporting document to the Guideline (GL) on Defined Approaches (DAs) for Skin Sensitisation- Annex 4. Organisation for Economic Cooperation and Development, Paris.

OECD (2021e). Series on Testing and Assessment No. 336: Supporting document to the Guideline (GL) on Defined Approaches (DAs) for Skin Sensitisation- Annex 6. Organisation for Economic Cooperation and Development, Paris.

Rovida, C., Basketter, D., Casati, S., de Silva, O. et al. (2007). Management of an integrated project (Sens-it-iv) to devel-op in vitro tests to assess sensitisation. Altern. Lab. Anim. 35, 317-22. doi:10.1177/026119290703500311

Takenouchi, O., Miyazawa, M., Saito, K., Ashikaga, T. et al. (2013). Predictive performance of the human Cell Line Activation Test (h-CLAT) for lipophilic chemicals with high octanol-water partition coefficients. J Toxicol Sci 38, 599-609. doi:10.2131/jts.38.599

Urbisch, D., Mehling, A., Guth, K., Ramirez, T. et al. (2015). Assessing skin sensitization hazard in mice and men using non-animal test methods. Regul. Toxicol. Pharmacol. 71, 337-51. doi:10.1016/j.yrtph.2014.12.008

Most read articles by the same author(s)

1 2 > >>