In vitro Method for Identifying Skin Sensitisers Combining Peptide Binding with ARE/EpRE Mediated Gene Expression in Human Skin Cells

Method Description

The SenCeeTox is an in vitro approach in which data on chemical reactivity towards Glutathione (GSH), gene expression profiling and cell viability are generated in order to identify the sensitisation potential of chemicals and to assign them into potency categories.

The method is addressing two biological mechanisms: a) the reactivity towards proteins and b) the activation of the ARE/EpRE signaling pathways in human keratinocytes, both of which are considered to be relevant steps in the inductions phase of skin sensitisation. Protein reactivity is measured in a cell-free system whereas gene induction is measured by qRT-PCR using mRNA isolated from 3D human epidermis models. Viability of cells is measured in parallel with the MTT assay. Such parameters are used in combination in a weighted algorithm to derive an in vitro toxicity index (IVTI) that is then used to predict a sensitiser potency category.