The relationship between CD86/CD54 expression and THP-1 cell viability in an in vitro skin sensitization test – human cell line activation test (h-CLAT)

Abstract

Recent regulations for cosmetics in Europe prohibit animal testing for evaluating the sensitization potential of chemicals to improve animal welfare. Yet, there is not an acceptable Organization for Economic Co-operation and Development non-animal skin sensitization test method. Several in vitro skin sensitization methods that focus on the activation of Langerhans cells, including human cell lines, are being evaluated as possible alternatives. In our previous study, we optimized our human cell line activation test (h-CLAT) using THP-1 cells (monocytic leukemia cell line) and conducted an inter-laboratory study. We found that measuring CD86/CD54 expression may be useful for predicting skin sensitization. The aim of this study was to confirm the relationship between CD86/CD54 expression and THP-1 cell viability in the h-CLAT. In this study, 21 allergens (e.g., dinitrochlorobenzene, p-phenylenediamine, Ni) and 8 non-allergens (e.g., SLS, lactic acid) were evaluated. For each chemical, more than 10 concentrations that gave a predicted cell viability range of 20-95% were used. The data showed that expression patterns of CD86/CD54 differed depending on chemical. For most allergens, cytotoxicity (65-90% cell viability) was needed for enhancement of CD86/CD54 expression. The criteria of "CD86 > or = 150 or CD54 > or = 200" resulted in an accuracy of 93%, which confirms appropriate cut-off criteria for h-CLAT. Furthermore, a good correlation was observed between EC3 of local lymph node assay and EC150(CD86) or EC200(CD54) of h-CLAT (12 or 16 chemicals, respectively), which would provide a useful estimate of allergic potency. These findings suggest that h-CLAT would be a good robust in vitro skin sensitization test.

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

“…However, despite promising data there are technical issues for the routine use of these cells such as donor variability, low cell numbers and availability of human blood (Rougier et al, 2000). To avoid such issues, human cell lines, with characteristics similar to DCs, such as THP-1 and MUTZ-3 cells have been utilised (Sakaguchi et al, 2009). MUTZ-3 comprises the most physiologically similar cell line to its in vivo DC counterpart and represents the immortalised equivalent of CD34 + DC precursors (Larsson et al, 2006).…”

Evaluation of Release and Accumulation of Metal Ions from Titanium and Nickel by Accelerated Dissolution Test in Simulated Body Environments

“…Therefore, we are now evaluating the sensitizing properties of metal ions by an in vitro method using human monocytic cells. 21,22 The information provided from this research may be utilized as an important starting point to estimate the allergenic properties of metallic biomaterials.…”

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.…”