冻存后人脐血DC介导的食管癌细胞瘤苗实验研究  

Experimental study of the cancer vaccine prepared by fusions of esophageal carcinoma cell line with dendritic cell derived from cord blood after cryopreservation

作　　者：陈素钻[1] 陈康文 俞晶[1] 谢丽华[1] 张娟[1] 罗丽莉[1] 郭光华[1] 

机构地区：[1]汕头大学医学院第一附属医院,广东汕头515041 [2]汕头市妇女儿童医院,广东汕头515041

出　　处：《现代肿瘤医学》2008年第7期1075-1079,共5页Journal of Modern Oncology

基　　金：广东省自然科学基金资助项目(编号:021228)

摘　　要：目的:探讨杂交瘤技术制备脐血树突状细胞(DC)和食管癌细胞的融合瘤苗在低温冻存后的生物学特性及特异性CTL活性。方法:分离脐血CD34+干细胞诱导扩增为成熟DC,与EC109细胞经聚乙二醇(PEG)法融合;免疫磁珠法筛选EC109-DC;鉴定瘤苗表型;-80℃低温冰箱冻存融合细胞EC109-DC,3周后融冻;观察融冻瘤苗表型及致瘤性;MTT法测定瘤苗诱导淋巴细胞增殖能力及体外特异性免疫应答能力。结果:融冻后疫苗体外半悬浮生长;同时高表达FR和CD80;融冻后瘤苗接种小鼠体内未见肿瘤形成;未冻存和冻存后融合疫苗EC109-DC活化的T淋巴细胞,可产生针对EC109细胞的特异性杀伤作用。结论:冻存未破坏融合疫苗的完整性;融冻后瘤苗无体内致瘤性,安全可靠;-80℃低温冰箱短期冻存对EC109-DC融合疫苗生物学特性及特异的CTL活性无明显影响,可望为DC疫苗提供简单可行的保存方法。Objective：To develop fusion vaccine of esophageal carcinoma cell and DC, and study the biological characteristics and induction antitumor immunity after cryopreservation. Methods： Fusion vaccine was produced by traditional PEG fusing, cytokine inducing, CD34^＋ magnetic microbead marker sorting, and MACS. The fused cells were frozen and stored at - 80 degrees C for three weeks. Both the flash vaccine and the cryopreservated vaccine were assayed for their immunophenotypes by flow cytometry. Oncogenicity of the cord blood derived DC based esophageal carcinoma vaccine after cryopreservation were tested. Lymphocytes proliferation reaction and the CTL cytotoxicity were examined with MTT assay both for the flash vaccine and the cryopreservated vaccine. Results ： The fusion cells after cryopreservation was expressed highly with FR and CD80. No tumor was found in spleen, lung and liver after the injection of fusion vaccine after cryopreservation ;The specific CTL activity was well preserved after cryopreservation, and there was no statistic significant difference when compared with that of the fresh fused vaccine （ P 〉 0.05 ）. Conclusion ： Cryopreservation does not cause significant changes in the phenotypes of the fused vaccine cells. The fusion vaccine after thawing have no oncogenicity in vivo. A simple method of freezing and storage at - 80℃ is practical. It can well preserve the phenotypes and morphology of the fusion cells. It will be a new storage method for cord blood derived DC -based vaccine.

关 键 词：低温保存 树突状细胞 食管癌细胞 细胞融合 癌症疫苗 

分 类 号：R730[医药卫生—肿瘤] R735.1[医药卫生—临床医学]