人外周血单核细胞释放PrP^C与其分泌Exosomes相关性研究  

Human Peripheral Blood Monocytes Release PrP^C in Association with Exosomes

作　　者：王桂花[1] 周向梅[1] 尹晓敏[1] 杨利峰[1] 杨建民[1] 赵德明[1] 

机构地区：[1]中国农业大学动物医学院,国家动物海绵状脑病实验室,北京100193

出　　处：《细胞生物学杂志》2009年第6期877-881,共5页Chinese Journal of Cell Biology

基　　金：国家科技支撑计划(No.2008BA154B06);国家自然科学基金(No.30871854)资助项目~~

摘　　要：为探求人外周血单核细胞是否分泌exosomes以及exosomes的分泌与细胞外环境中PrP^C的关系。采用密度梯度离心的方法从人外周血单核细胞系的培养基中提取exosomes,并利用电镜技术和免疫印迹分析对其形态学和生物学特征进行鉴定。结果显示所提取的exosomes主要分布在1.13～1.16 g/ml的密度层中,平均直径在30～100 nm之间,呈杯状,这与相关报道关于其他细胞分泌的exosomes特征分析的结果相一致;与exosomes相关的分子Tsg101、flotillin-1、Hsp70、PrP^C均为阳性标记,而作为内质网标志蛋白的钙连接蛋白在exosomes上没有检测到。以上结果说明人外周血单核细胞分泌的exosomes没有被细胞器污染,而且细胞向培养基中释放PrP^C与其分泌exosomes有关。In order to investigate whether human peripheral blood monocytes secrete exosomes, and exosomes associate with PrPC released into extracellular. Exosomes were isolated from the human peripheral blood monocytes medium by density gradient centrifugation, and their morphology and biological characteristics were identified by electron microscope and Western blot analysis. The results showed the density of fractions containing exosomes was between 1.13 and 1.16 g/ml. The exosomes were 30-100 nm in size and cup-shaped morphology, which are similar to the previous report for exosomes released from other cells. The associated molecules, Tsg101, flotillin-1, Hsp70, PrP^C were probed. While calnexin, a luminal marker of the endoplasmic reticulum, was not detected. We suggest that human peripheral blood monocytes can secrete exosomes without contaminated by organelles, and the release of PrPc associated with exosomes.

关 键 词：传染性海绵状脑病 细胞型朊蛋白 EXOSOMES flotillin-1 TSG101 

分 类 号：R742.9[医药卫生—神经病学与精神病学]