摘要
目的: 探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)与细胞膜微结构域脂筏的关系。方法: 采用间接免疫荧光流式细胞术, 分析K562细胞表面TRAIL分子的表达。用FITC标记的霍乱毒素B亚单位 (CTx)对K562细胞的脂筏进行染色; 用抗FITC单克隆抗体(mAb)交联脂筏后, 以兔抗TRAIL分子抗体及Cy3标记的羊抗兔抗体进行间接免疫荧光染色, 激光共聚焦显微镜分析TRAIL分子与脂筏微结构域的关系。结果: FITC -CTx和抗FITCmAb可使脂筏发生交联。脂筏交联的同时TRAIL分子发生聚集。动态观察表明, 抗FITC抗体作用 20min后, 脂筏发生交联, 作用 30min时交联最明显; 随着脂筏的交联, TRAIL分子发生聚集。抗FITC抗体作用 40min后, 脂筏交联程度减弱, 且TRAIL分子逐渐被排除于脂筏之外。结论: 抗FITC抗体可用于CTx-FITC脂筏染色后脂筏交联的研究。TRAIL分子可能与细胞膜脂筏微结构域有关。
AIM: To investigate the relationship between tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) and cell membrane microdomain lipid rafts. METHODS: The expression of TRAIL on K562 cells was detected by indirect immunofluorescence staining and flow cytometry. The lipid rafts on K562 cells were detected with FITC-labeled cholera toxin B subunit (CTx-FITC) which bound to the GM1 glycosphingolipid in lipid rafts. The lipid rafts were then cross-linked with an anti-FITC mAb after binding with CTx-FITC. The localization of TRAIL on K562 cells was analyzed with rabbit anti-TRAIL antibody, Cy3-labeled goat-anti-rabbit IgG (Cy3-labeled GAR) and laser scanning confocal microscope. RESULTS: The cross-linking of lipid rafts appeared at 20 minutes; reached peak at 30 minutes and weakened around 40 minutes. After the cells were labeled with CTx-FITC and anti-FITC mAb TRAIL was aggregated in the cross-linked lipid rafts. CONCLUSION: The anti FITC mAb could be used for cross-linking of lipid rafts. TRAIL can be recruited to lipid rafts.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2005年第1期1-5,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金青年科学基金资助项目(No.30200250 )
国家自然科学基金资助项目(No. 30371281)
