Hoya Masatoshi, Hirota Morihiko, Suzuki Mie, Hagino Shigenobu, Itagaki Hiroshi, Aiba Setsuya
Shiseido Quality Assessment Center, Shiseido Co Ltd, Yokohama-shi, Kanagawa, Japan.
Toxicol In Vitro. 2009 Aug;23(5):911-8. doi: 10.1016/j.tiv.2009.03.010. Epub 2009 Apr 7.
In this study, with the aim of developing a cell-based in vitro photosensitization assay, we examined whether changes of CD86 and CD54 expression on cells of a human monocytic cell line, THP-1, could be used to assess the photosensitizing potential of chemicals. First, we identified suitable conditions of UV-irradiation (irradiation dose; 5.0 J/cm(2), irradiation intensity; 1.7 mW/cm(2)) by investigating the effect of UV-irradiation on CD86 and CD54 expression on untreated or 6-methylcoumarin (a representative photoallergen)-treated THP-1 cells (irradiation method). However, acridine, a representative photo-irritant, augmented CD86 and CD54 expression on THP-1 cells, apparently via induction of reactive oxygen species (ROS). In order to abolish the effect of ROS, we examined CD86 and CD54 expression on THP-1 cells treated with pre-irradiated chemicals (pre-irradiation method). We found that UV-irradiated photoallergens, but not photo-irritants, enhanced CD86 and/or CD54 expression on the THP-1 cells. Finally, based on the results of irradiation, non-irradiation, and pre-irradiation with 18 test chemicals, we built a decision tree, which allows us to distinguish between photoallergens and photo-irritants. We suggest that this system may be useful for in vitro evaluation of the photoallergic potential of chemicals.
在本研究中,为了开发一种基于细胞的体外光致敏检测方法,我们研究了人单核细胞系THP-1细胞上CD86和CD54表达的变化是否可用于评估化学物质的光致敏潜力。首先,通过研究紫外线照射对未处理或经6-甲基香豆素(一种代表性光变应原)处理的THP-1细胞上CD86和CD54表达的影响(照射方法),我们确定了合适的紫外线照射条件(照射剂量;5.0 J/cm²,照射强度;1.7 mW/cm²)。然而,吖啶,一种代表性的光刺激剂,明显通过诱导活性氧(ROS)增强了THP-1细胞上CD86和CD54的表达。为了消除ROS的影响,我们检测了经预照射化学物质处理的THP-1细胞上CD86和CD54的表达(预照射方法)。我们发现紫外线照射的光变应原而非光刺激剂增强了THP-1细胞上CD86和/或CD54的表达。最后,基于18种测试化学物质的照射、未照射和预照射结果,我们构建了一个决策树,它使我们能够区分光变应原和光刺激剂。我们认为该系统可能有助于体外评估化学物质的光变应潜力。