树突状细胞分泌的exosomes体外诱导特异性CTL抗肾癌的初步研究  

A Preliminary Study on Antitumor Effect of Specific Cytotoxic T Lymphocytes Activated by Exosomes Secreted from Dendritic Cell Against Renal Carcinoma

作　　者：王新利[1] 李卿[1] 高婷[1] 付立叶[1] 王扬[1] 蒋涛[1] 姜又红[1] 

机构地区：[1]中国医科大学附属第一医院肿瘤研究所第二研究室,沈阳110001

出　　处：《中国医科大学学报》2010年第1期4-6,共3页Journal of China Medical University

基　　金：辽宁省自然科学基金资助项目(20042093)

摘　　要：目的提取树突状细胞外来体(DC)分泌的外来体(exosomes),检测其体外刺激同源T细胞活化和诱导特异性细胞毒性T细胞(CTL)抗肾癌769-P的免疫效应。方法从健康志愿者外周血单个核细胞中诱导培养DC,流式细胞术检测DC的表型;通过超速离心法结合超滤法制备DC分泌的exosomes,透射电镜观察exosomes的形态,MTT法检测DC分泌的exosomes诱导T细胞的增殖和特异性CTL对769-P的体外杀伤活性。结果超速离心法结合超滤法能从DC上清中分离exosomes,并且肿瘤抗原致敏DC分泌的exosomes能有效促进T细胞的增殖及特异性CTL抗769-P效应,T细胞增殖倍数达(1.68±0.22)%,CTL杀瘤活性为(38.23±2.16)%。结论抗原致敏DC分泌的exosomes能有效诱导激活具有较强杀瘤活性的肿瘤特异性CTL。Objective To extract exosomes from dendritic cell（DC）and investigate antitumor effect of the special cytotoxic T lymphocytes activated by exosomes against 769-P. Methods Monocytes from peripheral blood of healthy donors were cultured to induce DC in intro and their phenotypes were analyzed by FACS. The exosomes were extracted from DC by ultrafiltration and ultracentrifugation and observed under electric microscope. Proliferation of T cells and the cytotoxicities of CTL against 769-P induced by exosome were measured by MTT. Results The exosomes could be separated from culture supernatant of DC by ultrafiltration and ultracentrifugation. The combination of the exosomes from DC and the 769-P antigen could effectively stimulate T cell proliferation and enhance their cytotoxicity,the absorption value of T cell proliferation（1.68±0.22）,the cytotoxicity of CTL（38.23±2.16）%. Conclusion The exosomes extracted by ultrafiltration and ultracentrifugation can present tumor antigen to T cells and induce the cytotoxicity of CTL.

关 键 词：树突状细胞 外来体 细胞毒性T细胞 肾癌 

分 类 号：R730.51[医药卫生—肿瘤]