A comparative evaluation of in vitro skin sensitisation tests: the human cell-line activation test (h-CLAT) versus the local lymph node assay (LLNA).
We previously developed the human cell-line activation test (h-CLAT) in vitro skin sensitisation test, based on our reported finding that a 24-hour exposure of THP-1 cells (a human monocytic leukaemia cell line) to sensitisers is sufficient to induce the augmented expression of CD86 and CD54. The aim of this study is to confirm the predictive value of h-CLAT for skin sensitisation activity by employing a larger number of test chemicals. One hundred chemicals were selected, according to their categorisation in the local lymph node assay (LLNA), as being: extreme, strong, moderate and weak sensitisers, and non-sensitisers. The correlation of the h-CLAT results with the LLNA results was 84%. There were some false negatives (e.g. benzoyl peroxide, hexyl cinnamic aldehyde) and some false positives (e.g. 1-bromobutane, diethylphthalate). Eight out of the 9 false negatives (89%) were water-insoluble chemicals. The h-CLAT could positively predict not only extreme and strong sensitisers, but also moderate and weak sensitisers, though the detection rates of weak sensitisers and non-sensitisers were comparatively low. Some sensitisers enhanced both CD86 and CD54 levels, and some enhanced the level of only one of them. The use of the combination of CD86 and CD54 induction as a positive indicator, improved the accuracy of the test. In conclusion, the h-CLAT is expected to be a useful cell-based in vitro method for predicting skin sensitisation potential.
Ashikaga
Takao
T
Shiseido Co. Ltd, Quality Assessment Centre, Yokohama, Kanagawa, Japan. takao.ashikaga@to.shiseido.co.jp
Sakaguchi
Hitoshi
H
Sono
Sakiko
S
Kosaka
Nanae
N
Ishikawa
Makie
M
Nukada
Yuko
Y
Miyazawa
Masaaki
M
Ito
Yuichi
Y
Nishiyama
Naohiro
N
Itagaki
Hiroshi
H
D000826:Animal Testing Alternatives / Q000379:methods* / Q000592:standards; D000818:Animals; D015703:Antigens, CD / Q000187:drug effects / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D007114:Immunization; D023441:Local Lymph Node Assay; D008198:Lymph Nodes / Q000276:immunology; D009930:Organic Chemicals / Q000494:pharmacology; D011237:Predictive Value of Tests; D012867:Skin / Q000276:immunology; D012882:Skin Tests / Q000379:methods*
D015703:Antigens, CD; D009930:Organic Chemicals