As can be seen in Fig. 2B, the replicates of both 2-nitro-1,4-phenylendiamine GSI-IX clinical trial group closely together in a 2D Sammon projection, indicating a strong robustness and reproducibility of the assay. If triplicate samples
of any one stimulation end up on both sides of the hyperplane, it should be regarded as a sensitizer. Indeed, while the cutoff of a sample being a sensitizer or a non-sensitizer is currently set to zero, this cutoff should and will be evaluated in connection with pre-validation of the assay. Furthermore, a sample being ambiguously classified by the SVM is likely a weak sensitizer, as the absolute value of the decision value may be correlated to the potency of the sensitizer; the further away from the cloud of negative samples a sensitizer is positioned, the higher its potency
as a sensitizer, as discussed in (Johansson et al., 2011). Prediction of a compound’s ability to induce skin sensitization is an important aspect of safety assessment of chemicals, and is currently performed with animal models, such as the murine LLNA. However, a number of factors, such as the REACH legislation and the 7th amendment to the Cosmetics Directive, buy Ibrutinib make animal models unsuitable for assessment of sensitization. Furthermore, these assays are known to not correlate perfectly with clinical experience of human data. Indeed, the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) reported the accuracy of the LLNA to be 72% (Haneke et al., 2001). Genomic allergen rapid detection, GARD, is a novel assay for assessment of sensitization. It is based on a genomic readout,
measuring 200 transcripts in the myeloid cell line MUTZ-3 following compound stimulation. Lepirudin The 200 transcripts, collectively called GARD Prediction Signature, participate in signaling pathways that are involved with recognition of foreign substances. A number of these pathways, such as nuclear factor-erythroid-related factor 2 (NRF2) mediated oxidative response, aryl hydrocarbon receptor (AHR) signaling and Toll-like receptor (TLR) signaling, are known to lead to transcription of cytoprotective enzymes and DC maturation (Johansson et al., 2011) as a response to xenobiotic challenges. Thus, GARD utilizes human MUTZ-3 as an in vitro DC model, taking advantage of its decision-making role in the immune response leading to skin sensitization for predicting sensitizing potency in unknown chemicals. As a consequence of being an assay with a biomarker signature as readout, simultaneously monitoring a number of different cell events, GARD is well suited to detect positive compounds from a wide chemical space. The assay has been shown to be robust and highly reproducible, as well as accurate, with respect to the 38 reference compounds run so far.