热应激小鼠肝癌细胞(H_(22))源Exosomes的抗肿瘤免疫机制  

The mechanism of immune antitumor effect of HS-Exo derived from heat stressed mouse hepatoma cell line(H_(22))

作　　者：孙迪[1] 杨麟[1] 沈宜[1] 汪少华[1] 向自武[1] 

机构地区：[1]重庆医科大学基础医学院病理生理学教研室,重庆400016

出　　处：《复旦学报（医学版）》2009年第6期681-686,691,共7页Fudan University Journal of Medical Sciences

基　　金：重庆市渝中区科委基金项目

摘　　要：目的制备小鼠肝癌细胞(H22)源Exosomes及热应激小鼠肝癌细胞(H22)源HS-Exo(heat stressedExosomes,HS-Exo),研究其对小鼠可能的抗肿瘤免疫机制。方法用超速分级离心和蔗糖密度梯度离心纯化获得Exosomes及HS-Exo,透射电镜观察其形态。以Exosomes为对照,观察HS-Exo的蛋白组分、产量及其激发宿主抗肝癌免疫应答效应,用Western blot方法检测两者所含有的相关蛋白情况。用MTT法检测免疫小鼠脾细胞增殖和脾淋巴细胞的细胞毒活性。用免疫组化法检测经两者免疫后小鼠肿瘤组织中CD4+、CD8+淋巴细胞浸润情况。结果HS-Exo与Exosomes形态相似,HS-Exo所含的重要免疫蛋白表达增加(P<0.05),用HS-Exo免疫小鼠后比用Exosomes免疫小鼠能更有效地抑制肿瘤生长,更好地诱导淋巴细胞增殖,更显著增强小鼠脾淋巴细胞的细胞毒活性以及更明显的肿瘤治疗作用(P<0.05)。结论热应激制备HS-Exo的方法具有可行性,HS-Exo比Exosomes具有更强的免疫活性和肿瘤治疗作用。Objective To prepare Exosomes secreted by mouse hepatoma cell （H22） and heat stressed Exosomes （HS-Exo） derived from heat stress-treated mouse hepatoma cell （H22）, in order to study the possible anti-tumor immune mechanism. Methods Exosomes and HS-Exo were purified by serial ultracentrifugation and sucrose density gradiant centrifugation, and were observed and identified by electron microscope. The components and production of the protein and the effects of the host immune response against hepatocellular carcinoma of HS-Exo were observed by using Exosomes as the control. Their immunological factors were detected by Western blot. Lymphocyte proliferation and specific cytotoxie activity of mouse splenic cells were determined by MTT. CD4^＋ and CD8^＋ lymphoeytes infiltration in mouse tumor tissues immunized by both were analysed by immunohistochemical staining. Results HS-Exo was similar in morphology to the Exosomes, the important immune-ralated protein expressed in HS-Exo was increased （P〈0.05）. HS-Exo immunized mouse group showed more effective inhibition of tumor growth, better-induced lymphocyte proliferation,more significantly enhanced the cytotoxic activity of spleen lymphocytes, as well as a more prominent role in tumor therapy than Exosomes immunized mouse control group （P〈0.05）. Conclusions Heat stress treatment method for the preparation of HS-Exo was feasible. HS-Exo had a stronger role in the immuneactivity and tumor treatment than control Exosomes.

关 键 词：EXOSOMES 小鼠肝癌细胞(H22) HS-Exo 肿瘤免疫 

分 类 号：R363[医药卫生—病理学]