CAJ | 학술논문

目的体外诱导甲胎蛋白(AFP)158-166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50 ml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HLA-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T2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.
목적 체외유도갑태단백(AFP)158-166특이성T세포,검측기대AFP+간암세포적특이성살상능력.방법 채용류식세포술급PCR-SSP법종20례건강지원자중사선기인형HLA-A0201양성자,매례취외주혈50 ml획취단핵세포,세포인자첩벽점부배양수획수돌상세포(DC).장DC부재HLA-A0201표위태AFP158-166후안1∶10비례여신선림파세포혼합배양、증식,새서람법검측기대HepG2、부재화미부재AFP158-166적T2세포적살상능력.결과 성숙DC표면항원CD83、CD80화CD86양성솔분별위(81.3 ±2.4)％、(92.8±1.4)％화(70.5 ±1.9)％.유도적T세포재효파비40∶1시대HepG2、부재화미부재AFP158-166적T2세포살상솔분별위(61.1±3.4)％、(70.6±2.4)％화(16.3 ±2.7)％.결론 인외주혈체외유도적AFP158-166특이성T세포가살상AFP+ HepG2화부재AFP158-166적T2세포.
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 carriers were selected as blood donors by using flow cytometry and PCR-SSP from 20 healthy volunteers.Peripheral 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 AFP158-66 loaded T2 cells.