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Unsaturated compounds induce up-regulation of CD86 on dendritic cells in the in vitro sensitization assay LCSA

  • Immunotoxicology
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Abstract

Unsaturated compounds are known to cause false-positive reactions in the local lymph node assay (LLNA) but not in the guinea pig maximization test. We have tested a panel of substances (succinic acid, undecylenic acid, 1-octyn-3-ol, fumaric acid, maleic acid, linoleic acid, oleic acid, alpha-linolenic acid, squalene, and arachidonic acid) in the loose-fit coculture-based sensitization assay (LCSA) to evaluate whether unspecific activation of dendritic cells is a confounder for sensitization testing in vitro. Eight out of 10 tested substances caused significant up-regulation of CD86 on dendritic cells cocultured with keratinocytes and would have been classified as sensitizers; only succinic acid was tested negative, and squalene had to be excluded from data analysis due to poor solubility in cell culture medium. Based on human data, only undecylenic acid can be considered a true sensitizer. The true sensitizing potential of 1-octyn-3-ol is uncertain. Fumaric acid and its isomer maleic acid are not known as sensitizers, but their esters are contact allergens. A group of 18- to 20-carbon chain unsaturated fatty acids (linoleic acid, oleic acid, alpha-linolenic acid, and arachidonic acid) elicited the strongest reaction in vitro. This is possibly due to the formation of pro-inflammatory lipid mediators in the cell culture causing nonspecific activation of dendritic cells. In conclusion, both the LLNA and the LCSA seem to provide false-positive results for unsaturated fatty acids. The inclusion of T cells in dendritic cell-based in vitro sensitization assays may help to eliminate false-positive results due to nonspecific dendritic cell activation. This would lead to more accurate prediction of sensitizers, which is paramount for consumer health protection and occupational safety.

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Acknowledgments

The manuscript does not contain clinical studies or patient data. The LCSA is patented in Germany with the application number DE102007006736B4 (patent holder: Mrs. Dagmar Briechle). The authors have no financial relationship with the patent holder and declare no conflict of interest. Data presented in this publication are part of the doctoral thesis of Thomas Frohwein submitted to Charité – Universitätsmedizin Berlin. We thank Prof. Dr. med. Björn Dirk Krapohl for providing primary human keratinocytes for our experiments. We are grateful for a grant of the Sonnenfeld Stiftung, Berlin, providing financial support for the FACSCalibur.

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Authors and Affiliations

  1. Institute of Clinical Pharmacology and Toxicology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Thomas Armin Frohwein, Anna Sonnenburg, Ralf Stahlmann & Maximilian Schreiner

  2. Department of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Torsten Zuberbier

  3. Department of Internal Medicine (Abt. I), Bundeswehrkrankenhaus Berlin, Scharnhorststraße 13, 10115, Berlin, Germany

    Maximilian Schreiner

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  1. Thomas Armin Frohwein
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  2. Anna Sonnenburg
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  5. Maximilian Schreiner
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Correspondence to Maximilian Schreiner.

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Frohwein, T.A., Sonnenburg, A., Zuberbier, T. et al. Unsaturated compounds induce up-regulation of CD86 on dendritic cells in the in vitro sensitization assay LCSA. Arch Toxicol 90, 927–936 (2016). https://doi.org/10.1007/s00204-015-1527-4

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