小鼠肝癌细胞(H22)源外泌体的体内抗肝癌作用研究.docVIP

小鼠肝癌细胞(H22)源外泌体的体内抗肝癌作用研究 李 静，沈 宜，杨 麟，段 红，郑维平（重庆医科大学基础医学院病理生理教研室,重庆400016） 摘 要: 目的 在荷瘤动物模型观察小鼠肝癌细胞（H22）来源的外泌体(exosomes)激发宿主抗肝癌免疫应答的效应。方法 用自行分离纯化的小鼠肝癌细胞H22源外泌体免疫小鼠，然后给小鼠皮下注射H22肿瘤细胞，观察肿瘤的生长状况。用MTT法检测小鼠脾细胞增殖情况和细胞毒活性;用免疫组织化学法检测肿瘤组织中CD4+、CD8+ T淋巴细胞浸润情况。结果 该外泌体使肿瘤出现时间延迟，肿瘤生长缓慢，小鼠生存率提高。促进脾淋巴细胞增殖并增强其细胞毒活性，促进CD4+ 和CD8+ T细胞活化。结论 小鼠肝癌细胞（H22）源外泌体能诱导抗肿瘤应答，有效地诱导特异性的杀伤肿瘤细胞活性，有针对亲本细胞的免疫保护作用。 关键词: 外泌体；肝细胞癌；抗肿瘤免疫；肿瘤疫苗 中图法分类号： 文献标识码：A Antitumor effects of the exosomes derived from a mouse hepatoma carcinoma cell line(H22) LI Jing, SHEN Yi,YANG Lin, DUAN Hong, ZHENG Wei-ping. (Department of Pathophsiology, College of Basic Medicine, Chongqing University of Medical Sciences,Chongqing400016,China) Abstract: Objective To investigate the induction of antitumor response and its immune mechanism by H22 cell-derived exosomes. Methods The mice were immunized by H22 cell-derived exosomes and then inoculated with the wild type H22 finally the Tumor growth and long period survival were observed．Lymphocyte proliferation and specific cytotoxic activity of mice splenic cells was determined by MTT. Infiltration of lymphocytes in tumor tissues was analysied by immuno－histochemical staining of CD4+、CD8+ . Results Exosomes could induce lymphocyte proliferation.,generate strong specific cytotoxic spleen lymphocyte response and activate CD4+ and CD8+T cells both in tumor sites. Conclusion Vaccination of H22 cell-derived exosomes was effective in the induction of protective immunity. Those exosomes have significant antitumor activities． Key words: Exosomes; Hepatocellular carcinoma; antitumor immunity; tumor vaccine 肝癌是最常见的恶性肿瘤之一，在中国、日本、新加坡、韩国等东南亚国家发病率较高，近年来，西方国家的肝癌发病率也逐渐升高。肝癌对化疗和放疗均不敏感，易早期转移和术后复发，其他的治疗效果也不理想。需要进一步探求治疗与预防的方法。近年来，对肝癌的肿瘤免疫治疗、肿瘤疫苗方面进行研究和探讨成为一个新的发展方向。由多种细胞分泌的外泌体能将信息传递给抗原呈递细胞（APC）而引发较 作者简介：李 静，女，四川省德阳市人，硕士，讲师，主要从事肿瘤免疫方面的研究。电话：（023E-mail:lijing122510@ 通信作者：沈 宜，电话：（023E-mail: shenyicq@163.com 收稿日期：2007-09-14；修回日期：2007-11-18 强的免疫反应， 而且肿瘤细胞源外泌体作为一种新的肿瘤抗原携