体外诱导甲胎蛋白抗原表位肽特异性T细胞的研究  

In vitro induction of cefetoprotein antigen epitope a-fetoprotein158-166-specific T cells

作　　者：孙龙昊[1] 章由贤[1] 何向辉[1] 章志翔[1] 朱理玮[1] 

机构地区：[1]天津医科大学总医院普通外科,300052

出　　处：《中华实验外科杂志》2012年第11期2162-2164,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81172167）

摘　　要：目的体外诱导甲胎蛋白（AFP）156-166特异性T细胞，检测其对AFP＋肝癌细胞的特异性杀伤能力。方法采用流式细胞术及PCR-SSP法从20例健康志愿者中筛选基因型HLA-A0201阳性者，每例取外周血50ml获取单核细胞，细胞因子贴壁黏附培养收获树突状细胞（DC）。将DC负载HIA．A0201表位态AFP158-166后按1：10比例与新鲜淋巴细胞混合培养、增殖，噻唑蓝法检测其对HepG2、负载和未负载AFP158-166的T2细胞的杀伤能力。结果成熟DC表面抗原CD83、CD80和CD86阳性率分别为（81．3±2．4）％、（92．8±1．4）％和（70．5±1．9）％。诱导的T细胞在效靶比40：1时对HepG2、负载和未负载AFP158-166的眈细胞杀伤率分别为（61．1±3．4）％、（70．6±2．4）％和（16．3±2．7）％。结论人外周血体外诱导的AFP158-166特异性T细胞可杀伤AFP＋HepG2和负载AFP158-166的T2细胞。Objective To stimulate the α-fetoprotein （AFP）158-166-specific T cells in vitro and evaluate their specific cytotoxicity for AFP＋ hepatic carcinoma cells. Methods HLA-A0201 genotype carri- ers were selected as blood donors by using flow Cytometry and PCR-SSP from 20 healthy volunteers. Periph- eral blood mononuclear cells were harvested from 50 mL peripheral blood of each donor and cultured with cytokines to induce dendritic cells （DCs）. Donor lymphocytes were cultured with AFP158-166-loaded DCs at the ratio of 10： 1. Proliferative lymphocytes were collected and the cytotoxicity for hepatoma carcinoma cells （HepG2） , AFP158-166 loaded T2 cells and naive T2 cells was analyzed by using methyl thiazol tetrazolium （MTT） assay. Results The positive rate of DCs surface antigens CD83, CD80 and CD86 was （81.3± 2.4 ） %, （ 92. 8 ± 1.4 ） % and （ 70. 5 ± 1.9 ） % respectively. The cytotoxicity of AFP158-166-specific T cells to HepG2, AFP158-166 loaded T2 and naive T2 cells was （61.1±3.4）%, （70.6 ±2.4）% and （16.3 ± 2. 7 ） % respectively at the effector-target ratio of 40： 1. Conclusion The AFP158-166-specific T cells can be induced in vitro from the peripheral blood of healthy donors and have specific cytotoxicity for HepG2 and AFP158-166 loaded T2 ceils.

关 键 词：甲胎蛋白 树突状细胞 T细胞 癌 肝细胞 

分 类 号：R735.7[医药卫生—肿瘤]