肝素酶特异性细胞毒T淋巴细胞表位多抗原肽负载的树突状细胞疫苗体外抗肿瘤免疫效应研究  

Study on in vitro antitumor immune responses elicited by dendritic cells pulsed with heparanase multiple antigen peptides containing cytotoxic T lymphocyte epitopes

作　　者：王国珍[1] 汤旭东[1] 吴玉云[1] 李宁[1] 李长珠[1] 陈陵[1] 房殿春[1] 杨仕明[1] 

机构地区：[1]第三军医大学西南医院全军消化病研究所,重庆400038

出　　处：《第三军医大学学报》2011年第8期761-764,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30971341)~~

摘　　要：目的研究肝素酶(heparanase,Hpa)细胞毒T淋巴细胞(cytotoxic T lymphocyte,CTL)表位多抗原肽(multi-ple antigen peptides,MAP)疫苗能否诱导更强的Hpa特异性CTL反应。方法 HLA-A2.1限制性肝素酶CTL表位多抗原肽负载正常人外周血来源(peripheral blood mononuclear cell,PBMC)树突状细胞(dendritic cell,DC),诱导CTL,采用4 h标准51Cr释放实验检测上述CTL对不同肿瘤细胞的免疫杀伤效应;采用ELISPOT实验检测效应细胞分泌IFN-γ的能力。结果肝素酶CTL表位MAP疫苗诱导的CTL对Hpa阳性且HLA-A2.1匹配的KATO-Ⅲ胃癌细胞、U2OS骨肉瘤细胞、SW480结肠癌细胞具有杀伤效应,且其杀伤效应强于相应的单肽,在最大效应细胞/靶细胞(effector/target,E/T)时高出率均大于16%;其对HLA-A2.1阴性的HepG2肝癌细胞和Hpa阴性的MCF-7乳腺癌细胞不具有杀伤效应,但是对MCF-7/Hpa乳腺癌细胞和HepG2/HLA-A2肝癌细胞具有杀伤效应,且其杀伤效应强于相应的单肽,在最大E/T时高出率均大于18%;其对自体淋巴细胞和DC不具有杀伤效应。另一方面人肝素酶CTL表位MAP疫苗诱导的IFN-γ分泌水平强于相应的单肽。结论肝素酶CTL表位MAP疫苗能激发较相应单肽更强的特异性和非特异性抗肿瘤效应。Objective To study whether the multiple antigen peptides（MAPs） of heparanase（Hpa） can elicit stronger Hpa-specific cytotoxic T lymphocyte（CTL） responses against various tumors in vitro.Methods Dendritic cells（DCs） were pulsed with HLA-A2.1-restricted Hpa MAPs and Hpa single peptides to induce Hpa-specific CTLs.The lysis of KATO-Ⅲ gastric cancer cells,U2OS osteogenic sarcoma cells,SW480 colon cancer cells,HepG2 liver cancer cells,MCF-7 breast cancer cells,HepG2/HLA-A2.1 liver cancer cells,and MCF-7/Hpa breast cancer cells by Hpa-specific CTLs was examined by 4-h standard 51Cr release assay.The IFN-γ production of the effector cells was tested with ELISPOT.Results Compared with those induced by Hpa single peptides,the CTLs induced by Hpa MAPs had stronger specific lysis effects on Hpa-and HLA-A2.1-positive tumor cells（KATO-Ⅲ,U2OS,and SW480）,by more than 16% at the maximum effector/target（E/T） ratio.Neither Hpa-negative MCF-7 cells nor HLA-A2.1-negative HepG2 cells were lysed by the CTLs.The effector cells induced by Hpa MAPs,however,could lyse HepG2/HLA-A2.1 cells（HLA-A2.1-positive） and MCF-7/Hpa cells（Hpa-positive）,and had stronger specific lysis effects than those induced by Hpa single peptides,by more than 18% at the maximum E/T ratio.Hpa MAPs could not elicit lysis of auto-logous lymphocytes and DCs.Compared with Hpa single peptides,Hpa MAPs could increase the frequency of IFN-γ-producing T cells.Conclusions This study shows that Hpa MAPs can elicit stronger specific and nonspecific antitumor immune responses by Hpa-specific CTLs.

关 键 词：肝素酶 CTL表位 多抗原肽 树突状细胞 肿瘤疫苗 

分 类 号：R73-362[医药卫生—肿瘤] R392.13[医药卫生—临床医学]