Department of Microbiology and Immunology, and Institute of Endemic Disease, College of Medicine, Seoul National University, Seoul, 110-799, South Korea.
Department of Occupational Health, College of Natural Sciences, Catholic University of Daegu, Daegu, 712-702, South Korea.
Toxicol In Vitro. 2018 Feb;46:304-312. doi: 10.1016/j.tiv.2017.10.018. Epub 2017 Oct 18.
Due to considerable constraints in using animals for risk assessment, much effort has been directed at developing non-animal test methods. Developing assays for skin sensitization, the leading cause of contact dermatitis, is particularly important, but there are currently no in vitro skin sensitization tests that completely replace animal tests. HaCaSens, a simple skin sensitization test using non-transformed HaCaT cells, predicts keratinocyte activation by skin sensitizers with 75% sensitivity, 83% specificity and 77% accuracy in a previous study using 22 coded substances. Although the data show promising results, the number of tested substances is insufficient to prove predictive capacity. Moreover, reproducibility among different laboratories has not been studied. Here, three laboratories participated in a validation in order to assess HaCaSens feasibility for official validation. To examine transferability, intra- and inter-lab reproducibility and predictive capacity, HaCaSens was assessed on a set of 30 test substances coordinated by the Validation Management Team (VMT). The results showed satisfactory transferability as well as intra- and inter-laboratory reproducibility. Further assessment of its predictive capacity on 20 test substances demonstrated a sensitivity of 81.8% (18/22), specificity of 87.5% (7/8), and accuracy of 83.3% (25/30) in identifying skin sensitizers, which is comparable with presently validated assays, KeratinoSens™ and LuSens. This validation study shows that the HaCaSens assay is easily transferable, reproducible and highly predictable for identifying skin sensitizers.
由于在风险评估中使用动物存在相当大的限制，因此人们投入了大量精力来开发非动物测试方法。开发皮肤致敏性的检测方法尤为重要，皮肤致敏性是接触性皮炎的主要原因，但目前还没有完全替代动物测试的体外皮肤致敏性测试方法。HaCaSens 是一种使用非转化 HaCaT 细胞的简单皮肤致敏性测试方法，在前一项使用 22 种编码物质的研究中，该测试方法对皮肤致敏剂诱导角质形成细胞激活的敏感性为 75%，特异性为 83%，准确性为 77%。尽管数据显示出有希望的结果，但测试物质的数量不足以证明其预测能力。此外，不同实验室之间的重现性尚未得到研究。在这里，三个实验室参与了验证，以评估 HaCaSens 用于正式验证的可行性。为了研究可转移性、实验室内部和实验室之间的重现性以及预测能力，由验证管理小组（VMT）协调的 30 种测试物质对 HaCaSens 进行了评估。结果表明，该方法具有令人满意的可转移性以及实验室内部和实验室之间的重现性。进一步对 20 种测试物质的预测能力进行评估表明，该方法对皮肤致敏剂的识别具有 81.8%的敏感性（18/22）、87.5%的特异性（7/8）和 83.3%的准确性（25/30），与目前经过验证的 KeratinoSens™ 和 LuSens 检测方法相当。这项验证研究表明，HaCaSens 检测方法易于转移、重现性高且对识别皮肤致敏剂具有高度预测性。
