Development and validation of a new in vitro assay designed to measure contact allergen-triggered oxidative stress in dendritic cells.
Development and validation of a new in vitro assay designed to measure contact allergen-triggered oxidative stress in dendritic cells.
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DOI：
10.1016/j.jdermsci.2012.08.003
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发表时间：
2012-11
影响因子：
4.6
通讯作者：
Takashima A
中科院分区：
文献类型：
--
作者：
Miyazawa M;Takashima A
已知选择的接触性过敏原诱导树突状细胞（DC）的表型和功能成熟。DC中发生的这种变化已被用作测定读数来预测小化学品的皮肤致敏潜力。为了满足对可靠的体外试验以识别接触性过敏原的迫切需求，我们试图开发一种基于DC的检测方法，旨在检测致敏剂诱导的早期变化。采用基因芯片技术和定量RT-PCR技术检测小鼠皮肤和XS 106 DC细胞暴露于二硝基氟苯（DNFB）后的基因表达谱。通过测量氧化应激水平间接检查活性氧物质（ROS）的产生-用荧光染料CM-H2 DCFDA标记XS 106 DC，暴露于测试化学品，然后通过流式细胞仪检查荧光信号。DNFB诱导小鼠皮肤和XS 106 DC中多种氧化还原调节基因的mRNA表达。这些基因的表达是诱导过氧化氢和ROS抑制剂，diphenyleneiodonium阻断。快速和显着的活性氧产生诱导的25个测试的28种皮肤致敏剂，但只有3个测试的21种皮肤刺激物。我们的小规模验证研究证明了我们的基于DC的ROS产生测定法在检测具有不同致敏效力的结构多样的接触性过敏原方面的实用性。推测DCs中ROS的产生可能是致敏阶段的早期事件是很有吸引力的。
Selected contact allergens are known to induce phenotypic and functional maturation of dendritic cells (DCs). Such changes occurring in DCs have been employed as assay readouts to predict skin-sensitizing potentials of small chemicals. To respond to the urgent needs for reliable in vitro tests to identify contact allergens, we sought to develop a DC-based assay designed to detect early change(s) induced by sensitizers. Signature gene expression profiles of skin sensitization were determined by GeneChip and quantitative RT-PCR analyses of RNA samples harvested from mouse skin and XS106 DC line after exposure to dinitrofluorobenzene (DNFB). Production of reactive oxygen species (ROS) was examined indirectly by measuring the level of oxidative stress – XS106 DCs were labeled with a fluorescent dye, CM-H2DCFDA, exposed to test chemicals, and then examined for fluorescence signals by flow cytometer. DNFB induced abundant mRNA expression of several redox regulatory genes in both mouse skin and XS106DCs. Expression of these genes was inducible by hydrogen peroxide and blocked by a ROS inhibitor, diphenyleneiodonium. Rapid and significant ROS production was induced by 25 of the 28 tested skin sensitizers, but only by 3 of the 21 tested skin irritants. Our small-scale validation study demonstrates the practical utility of our DC-based ROS production assay to detect structurally diverse contact allergens with varying sensitizing potencies. It is tempting to speculate that ROS production in DCs may represent an early event during the sensitization phase.
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DOI：
10.1073/pnas.172398899
发表时间：
2002-09-03
影响因子：
11.1
作者：
Dinkova-Kostova, AT;Holtzclaw, WD;Talalay, P
通讯作者：
Talalay, P
影响因子：
3.8
作者：
Gerberick, GF;Vassallo, JD;Lepoittevin, JP
通讯作者：
Lepoittevin, JP
影响因子：
6.5
作者：
Bergstrom, Moa Andresen;Ott, Hagen;Baron, Jens M.
通讯作者：
Baron, Jens M.
影响因子：
6.5
作者：
Gildea, Lucy A.;Ryan, Cindy A.;Gerberick, G. Frank
通讯作者：
Gerberick, G. Frank
影响因子：
82.9
作者：
Matsue, H;Matsue, K;Takashima, A
通讯作者：
Takashima, A