Comparison and validation of an in vitro skin sensitization strategy using a data set of 33 chemical references

Allergic contact dermatitis (ACD) is an adverse health effect that develops following repeated exposure to skin sensitizing chemicals. An animal testing ban has been applied in EU, leading to development of reliably predictive non-animal methods. Several in vitro methods have been developed as alternatives but one single non-animal test method is not been sufficient to fully address since the LLNA test ban. Here, we have selected an ITS (Integrated Testing Strategy) for skin sensitization which focuses on three in vitro methods that covered the first three steps of the AOP (DPRA, SENS-IS or h-CLAT). The aim of this study was to compare these three methods due to the WoE approach based on a 2-out-of-3-assessment. The results of 33 references were compared to in vivo data (especially human). We have shown that tested firstly DPRA and SENS-IS have permitted to conclude on 29 of 33 chemicals, whereas DPRA and h-CLAT on 25, and SENS-IS and h-CLAT on 23. With this sequence, DPRA and SENS-IS and then h-CLAT in case of equivocal results, we conclude more quickly by performing fewer tests. Thereby, we have shown that it is better to follow a preferential sequence than testing chemicals simultaneously with these three methods.