Validation of the GARD™skin assay for assessment of chemical skin sensitizers – ring trial results of predictive performance and reproducibility

Toxicological Sciences, kfz108,

Henrik Johansson, Robin Gradin, Angelica Johansson, Els Adriaens, Amber Edwards, Veronika Zuckerstätter, Anders Jerre, Florence Burleson, Helge Gehrke, Erwin Roggen

Proactive identification of chemicals with skin sensitizing properties is a key toxicological endpoint within chemical safety assessment, as required by legislation for registration of chemicals. In order to meet demands of increased animal welfare and facilitate increased testing efficiency also in non-regulatory settings, considerable efforts have been made to develop non-animal approaches to replace current animal testing.

Genomic Allergen Rapid Detection (GARD) is a state-of-the-art technology platform, the most advanced application of which is the assay for assessment of skin sensitizing chemicals, GARDskin. The methodology is based on a dendritic cell (DC)-like cell line, thus mimicking the mechanistic events leading to initiation and modulation of downstream immunological responses. Induced transcriptional changes are measured following exposure to test chemicals, providing a detailed evaluation of cell activation. These changes are associated with the immunological decision-making role of DCs in vivo and include among other phenotypic modifications, up-regulation of co-stimulatory molecules, induction of cellular and oxidative stress pathways and xenobiotic responses and provide a holistic readout of substance-induced DC activation.Here, results from an inter-laboratory ring trial of GARDskin, conducted in compliance with OECD guidance documents and comprising a blinded chemical test set of 28 chemicals, are summarized. The assay was found to be transferable to naïve laboratories, with an inter-laboratory reproducibility of 92.0%. The within-laboratory reproducibility ranged between 82.1-88.9%, while the cumulative predictive accuracy across the three laboratories was 93.8%.It was concluded that GARDskin is a robust and reliable method for the identification of skin sensitizing chemicals and suitable for stand-alone use or as a constituent of integrated testing. These data form the basis for the regulatory validation of GARDskin.
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