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超顺磁性氧化铁标记自然调节性T细胞体外研究

发表时间:2012-06-15  浏览量:1156  下载量:364
全部作者: 陈唯唯,余道武,熊伟,穆可涛,漆剑频,朱文珍
作者单位: 华中科技大学同济医学院附属同济医院放射科;华中科技大学同济医学院附属同济医院器官移植研究所
摘 要: 目的:体外采用磁共振(magnetic resonance,MR)对比剂超顺磁性氧化铁(superparamagnetic iron oxide,SPIO)对调节性T细胞(regulatory T cells,Treg)进行标记,研究SPIO对调节性T细胞的生物学影响,为SPIO对调节性T细胞体内回输治疗的磁共振监测提供依据。方法:采用试剂盒分离纯化小鼠自然调节性T细胞。将分离的调节性T细胞与浓度分别为0.01,0.05,0.10,0.15 g/L的二巯基丁二酸(DMSA)包被的SPIO共同孵育,采用普鲁士蓝染色和原子吸收分光光度计法检测调节性T细胞对SPIO的摄取情况。采用噻唑蓝(MTT)方法检测标记后的调节性T细胞的存活情况。通过流式细胞仪检测Foxp3的表达状况,了解标记后的调节性T细胞的表型变化。通过检测对淋巴细胞的抑制增殖作用了解标记后调节性T细胞的功能变化。对琼脂凝胶标记后的细胞重悬液进行磁共振扫描,了解标记后细胞的信号变化。结果:调节性T细胞可摄取SPIO,SPIO浓度越大,摄取SPIO量越大;在一定浓度范围内,SPIO对调节性T细胞存活率无明显影响,可使调节性T细胞保持表型,同时维持一定的免疫调节功能。MR扫描图像显示SPIO标记的细胞T2加权相信号明显降低。结论:DMSA包被的SPIO体外可以成功标记调节性T细胞。在一定浓度范围内,SPIO对调节性T细胞的表型、活力、免疫调节作用无明显影响。
关 键 词: 医学影像学;超顺磁性氧化铁;磁共振成像;调节性T细胞;体外实验
Title: Preliminary study on labeling natural regulatory T cells with superparamagnetic iron oxide in vitro
Author: CHEN Weiwei, YU Daowu, XIONG Wei, MU Ketao, QI Jianpin, ZHU Wenzhen
Organization: Department of Radiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology; Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology
Abstract: Objective: To label the regulatory T cells with superparamagnetic iron oxide (SPIO) and evaluate the biological effects of SPIO on the regulatory T cells in vitro. Methods: The natural regulatory T cells were isolated from the spleen of mice with EasySep Mouse CD4+ T cell pre- enrichment kit and Mouse CD25 Selection kit. The cells were then incubated with SPIO for 2 h at the iron concentration of 0.01, 0.05, 0.10 and 0.15 g/L. Cells incubated without SPIO served as the control. The uptake capability of SPIO was evaluated by Prussian blue staining and measured quantitatively with atomic absorption spectrometry. The cell viability was monitored by MTT method. The suppressive effect of regulatory T cells on the proliferation of CD4+ T cells was analyzed in proliferation assays. Results: SPIO was uptaken by regulatory T cells in a concentration- dependence manner. Within a certain range of the concentration, SPIO had no obvious effects on the viability, phenotype and suppressive function of the regulatory T cells. The MR images showed the significant decrease of magnetic resonance (MR) signal in T2 weighted images resulted from SPIO labeling of cells. Conclusion: the regulatory T cells were successfully labeled with SPIO in vitro. In a certain range of concentration, SPIO have no effects on phenotype, viability and function of regulatory T cells.
Key words: medical imaging; superparamagnetic iron oxide; magnetic resonance imaging; regulatory T cells; in vitro
发表期数: 2012年6月第11期
引用格式: 陈唯唯,余道武,熊伟,等. 超顺磁性氧化铁标记自然调节性T细胞体外研究[J]. 中国科技论文在线精品论文,2012,5(11):1039-1046.
 
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