尤文肉瘤细胞总RNA转染的DC疫苗体外诱导尤文肉瘤患者特异性抗肿瘤免疫的实验研究  

Experimental Study of Specific Antitumor Immunity Induced by Dendritic Cell Vaccine Transfected with Tumor Cell Total RNA in Ewing Sarcoma Patients

作　　者：王剑[1] 桑宏勋[1] 王臻[1] 郭征[1] 

机构地区：[1]第四军医大学西京医院骨科研究所,西安710033

出　　处：《科学技术与工程》2006年第13期1772-1776,共5页Science Technology and Engineering

基　　金：国家自然科学基金(39770749)资助

摘　　要：为了探讨尤文肉瘤细胞总RNA转染的DC疫苗体外诱导特异性抗肿瘤免疫的能力,采用分离尤文肉瘤患者外周血单核细胞体外诱导DC细胞,Trizol法提取患者尤文肉瘤细胞总RNA,用总RNA转染DC并诱导特异性CTL的扩增,用RTPCR方法检测肿瘤总RNA加载的DC细胞的表达,用MTT法检测淋巴细胞的增殖和CTL的杀伤活性。结果RTPCR测定显示尤文肉瘤细胞总RNA转染的DC细胞可以呈递肿瘤特异性抗原,并特异性地表达其特有序列EWSFLI1。经尤文肉瘤细胞总RNA转染的DC特异性表面标志及功能相关分子表达均上调,转染后的DC可显著刺激自体T淋巴细胞增殖。诱导的特异性CTL对携带EWSFLI1抗原的靶细胞的杀伤率显著高于LAK细胞和未经转染的DC。说明尤文肉瘤细胞总RNA转染的DC疫苗可在体外诱导出特异性抗肿瘤免疫。To observe ability of DC vaccine transfected with tumor cell total RNA to induce specific antitumor immunity in Ewing sarcoma patients in vitro, DCs were generated from Ewing＇ s sarcoma patients＇peripheral blood mononuclear cells（PBMC）. Total RNA was isolated from Ewing＇s sarcoma cell by Trizol. Autologous DCs transfected with total RNA of Ewing＇s sarcoma cell were used to induce specific CTL proliferation. The expression of DCs tansfected with tumor total RNA was measured by RT-PCR method. T cells pro- liferation and specific cytotoxicity were measured by MTT method. RT-PCR shows that DCs pulsed with Ewing＇sarcoma cells total RNA can present tumor specific antigen and express its characteristic sequence EWS-FLI1 specificly. Transfected DCs exhibited dramatically increased expression of specific membrane markers and function-associated molecules, and were more potent in stimulating autologous T cell proliferation than that of control DCs. Specific CTLs induced by transfected DCs showed higher cytotoxicity than LAK and control DCS against EWS-FLI1 positive Ewing＇sarcoma cells. It is conclused that Ewing＇s sarcoma patients＇autologous DCs transfected with tumor total RNA are effective vaccines in stimulating specific antitumor T-cell immunity in vitro.

关 键 词：树突状细胞 尤文肉瘤 RNA 肿瘤免疫 疫苗 

分 类 号：R392.9[医药卫生—免疫学]