肝素酶CTL表位多抗原肽（MAP）负载的树突状细胞疫苗体外抗肿瘤免疫效.docVIP

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 王国珍，汤旭东，吴玉云，李 宁，李长珠，陈 陵，房殿春，杨仕明 （400038重庆，第三军医大学西南医院全军消化病研究所） [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（multiple 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。 [关键词]肝素酶；CTL表位；多抗原肽；树突状细胞；肿瘤疫苗 Heparanase CTL epitope MAP-pulsed DCs elicit immune responses against various tumors in vitro WANG Guo-zhen，TANG Xudong，WU Yuyun， LI Nin， LI Changzhu，CHEN ling， FANG Dianchun，YANG Shiming（Institute of Gastoenterology， Southwest Hospital， The Third Miltary Medical University， Chongqing， 400038） [ABSTRACT] Objective The present paper is designed to study whether the heparanase multiple antigen peptide could elicit stronger heparanase-specific cytotoxic T lymphocytes （CTLs） against various tumors in vitro. Methods Dendritic cells （DCs） were pulsed with HLA-A2.1-restricted multiple antigen peptide of heparanase to induce heparanase-specific CTLs. Specific lysis to various tumor cell lines was examined by a 4 h standard 51Cr release assay. We use ELISPOT to test IFN-γ–producing of the effector cell. Results comparing to Heparanase epitope peptide，Heparanase multiple antigen peptide could elicit stronger specific lysis to gastric cancer cells KATO-III， osteogenic sarcoma cells U2OS and colonic cancer cells SW480， which were heparanase and HLA-A2.1positive, at the highest E/T the hyper-rate is grater than 16 percent.Effector induced by the Heparanase multiple antigen peptide neighter lyse MCF-