Byamba Dashlkhumbe, Kim Tae Gyun, Kim Dong Hyun, Je Jeong Hwan, Lee Min-Geol
Department of Dermatology and Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea.
Ann Dermatol. 2010 Aug;22(3):269-78. doi: 10.5021/ad.2010.22.3.269. Epub 2010 Aug 5.
Although reactive oxygen species (ROS) have been produced in both mouse bone marrow-derived dendritic cells (DCs) and XS-106 DCs by contact sensitizers and irritants in previous studies, the generation of ROS in human monocyte-derived DCs (MoDCs) and their role in contact hypersensitivity (CHS) has yet to be elucidated.
The purpose of this study was to determine whether contact allergens and irritants induce ROS in MoDCs and, if so, to evaluate the role of contact allergen and irritant induced-ROS in MoDCs in CHS.
Production of ROS was measured by 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H(2)DCFDA) assay. Surface CD86 and HLA-DR molecules were detected by flow cytometry. Protein carbonylation was detected by Western blotting.
ROS were produced by contact allergens such as dinitrochlorobenzene (DNCB) and thimerosal and the irritant benzalkonium chloride (BKC). DNCB-induced, but not BKC-induced, ROS increased surface CD86 and HLA-DR molecules on MoDCs and induced protein carbonylation. These changes were reduced in the presence of antioxidant N-acetyl cysteine.
Our results suggest that DNCB-induced ROS may be different from those induced by irritant BKC. The DNCB-induced ROS may be associated with the CHS response, because they activate surface molecules on DCs that are important for generating immune reactions.
尽管在先前的研究中,接触性致敏剂和刺激物已在小鼠骨髓来源的树突状细胞(DCs)和XS - 106 DCs中产生了活性氧(ROS),但人类单核细胞来源的树突状细胞(MoDCs)中ROS的产生及其在接触性超敏反应(CHS)中的作用尚未阐明。
本研究的目的是确定接触性变应原和刺激物是否会在MoDCs中诱导ROS产生,如果是,则评估接触性变应原和刺激物诱导的ROS在MoDCs的CHS中的作用。
通过5 -(和 - 6)- 氯甲基 - 2',7' - 二氯二氢荧光素二乙酸酯(CM - H₂DCFDA)测定法测量ROS的产生。通过流式细胞术检测表面CD86和HLA - DR分子。通过蛋白质印迹法检测蛋白质羰基化。
接触性变应原如二硝基氯苯(DNCB)、硫柳汞和刺激物苯扎氯铵(BKC)可产生ROS。DNCB诱导而非BKC诱导的ROS增加了MoDCs表面的CD86和HLA - DR分子,并诱导了蛋白质羰基化。在抗氧化剂N - 乙酰半胱氨酸存在下,这些变化减少。
我们的结果表明,DNCB诱导的ROS可能与刺激物BKC诱导的ROS不同。DNCB诱导的ROS可能与CHS反应相关,因为它们激活了DCs上对产生免疫反应很重要的表面分子。