Development of an in vitro skin sensitization test using human cell lines; human Cell Line Activation Test (h-CLAT) II. An inter-laboratory study of the h-CLAT

Identification of PDL-1 as a novel biomarker of sensitizer exposure in dendritic-like cells

“…Myriad reports have highlighted CD86 as a suitable marker for developing an in vitro test system (Aiba et al, 1997;Tuschl and Kovac, 2001;De Smedt et al, 2002;Aeby et al, 2004;Boisleve et al, 2004;Sakaguchi et al, 2006;Pepin et al, 2007), although Hulette et al (2005) reported that CD86 was an unreliable biomarker in primary DCs due to differences in donor variability and responsiveness. In contrast to these findings, in the present study, CD86 was found to be consistently and specifically up-regulated on MUTZ-3 cells in response to DNCB, Cin and PPD, although, moDCs only showed a response to the sensitizers DNCB and E. Thus, although CD86 may be a useful biomarker, marked differences were observed in the phenotypic responses to the sensitizers between the two cell types.…”

“…In contrast to these findings, in the present study, CD86 was found to be consistently and specifically up-regulated on MUTZ-3 cells in response to DNCB, Cin and PPD, although, moDCs only showed a response to the sensitizers DNCB and E. Thus, although CD86 may be a useful biomarker, marked differences were observed in the phenotypic responses to the sensitizers between the two cell types. Sakaguchi et al (2006) also observed variable responses depending on both the sensitizer and cell type used suggesting that sensitizers act differently depending on the cellular system (Sakaguchi et al, 2006). Recent work has indicated that the cell responsiveness to sensitizers is affected by external 'danger signals' (Lavergne et al, 2009) and hapten:protein interactions (Jenkinson et al, 2009) hours (Goldstein et al, 1996;Pype et al, 1999).…”

Keratinocytes, Innate Immunity and Allergic Contact Dermatitis - Opportunities for the Development of In Vitro Assays to Predict the Sensitizing Potential of Chemicals

“…Both assays apply chemicals to a monocytic cell line, U937 and THP-1 respectively, and measure the upregulation of CD86 protein on the cell surface. In addition to CD86, the upregulation of CD54 is measured in the h-CLAT assay (Sakaguchi et al, 2006;Sakaguchi et al, 2007;Sakaguchi et al, 2009;Ashikaga et al, 2010). Both assays are currently in ECVAM prevalidation.…”

An economic approach to non-animal toxicity testing for skin sensitisation

“…Activation Test (h-CLAT) Sakaguchi et al, 2006;Ashikaga et al, 2010;Sakaguchi et al, 2010), which have been validated by the European Centre for Validation of Alternative Methods (ECVAM; Italy). The DPRA is described in the testing guideline (TG)…”

“…Of these, one in chemico method and two in vitro methods were formally validated, i.e. the Direct Peptide Reactivity Assay (DPRA; (Gerberick et al, 2007)), the human Cell-Line Activation Test (h-CLAT; (Sakaguchi et al, 2006)) and the ARE-Nrf2 luciferase method covered by KeratinoSens TM (Emter et al, 2010).…”