表达EBV LMP2的小鼠骨髓间充质干细胞免疫原性初讨  

A preliminary study on the immunological effect of mouse mesenchymal stem cells expressing EBV LMP2

作　　者：孙立莹[1] 杜海军[2] 李红霞[2] 叶树清[2] 周玲[2] 曾毅[2] 王湛[2] 

机构地区：[1]北京工业大学生命与生物工程学院,北京100022 [2]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,北京100052

出　　处：《中国病毒病杂志》2015年第3期201-207,共7页Chinese Journal of Viral Diseases

基　　金：国家高技术研究发展计划(863计划)项目(2012AA02A404);国家重点基础研究发展计划(973计划)项目(2011CB504800);环境与病毒肿瘤学北京市重点实验室开放课题(EV02012021)

摘　　要：目的贴壁法分离小鼠骨髓间充质干细胞(MSCs),观察表达EBV LMP2蛋白的小鼠MSCs(MSCs-LMP2)体内诱导特异性CTL应答效果。方法贴壁培养法分离BALB/c小鼠MSCs,流式细胞术鉴定MSCs纯度。利用携带EBV LMP2基因的非复制重组5型腺病毒(Ad5-LMP2)感染已鉴定小鼠MSCs,制备表达EBV LMP2蛋白的MSCs-LMP2。使用MSCs-LMP2肌内注射的方式免疫BALB/c小鼠,每次免疫剂量6×105个细胞,免疫间隔为1周,连续免疫5次。末次免疫后1周分离小鼠脾淋巴细胞,Elispot方法检测EBV LMP2特异性CTL应答。结果贴壁培养法分离、纯化的小鼠骨髓MSCs纯度可达91.8%;Ad5-LMP2感染的MSCs能很好表达LMP2蛋白,形成MSCs-LMP2,MSCs-LMP2免疫同种小鼠后,在1×106个小鼠脾淋巴细胞中,诱导产生的LMP2特异性CTL能达到323个。结论贴壁培养法能够获得纯度较高的小鼠骨髓MSCs;LMP2修饰的MSCs(MSCs-LMP2)能诱导同种小鼠产生特异性CTL应答。Objective To isolate mouse mesenchymal stem cells（MSCs）using adherent culture and observe the specific CTL response induced in mouse in vivo by MSCs expressing EBV LMP2（MSCs-LMP2）. MethodsBALB/c mouse MSCs were isolated using adherent culture and the purity of MSCs obtained was determined with flow cytometry.The purified mouse MSCs were infected with a replication-deficient recombinant adenovirus carrying the EBV LMP2gene（Ad5-LMP2）to prepare MSCs-LMP2 expressing EBV LMP2.BALB/c mice were immunized through intramuscular injection of MSCs-LMP2,with a dose of 6×105 cells/mouse each time,at an interval of 1week,and for 5times in total.One week after the last immunization,the mouse splenocytes were isolated and EBV LMP2-specific CTL was detected using Elispot assay. Results The purity of MSCs obtained from the isolation and purification of mouse MSCs using adherent culture was as high as 91.8%.MSCs infected by Ad5-LMP2 were able to well express LMP2 and constitute MSCs-LMP2.After immunizing the same strain of mice with MSCs-LMP2,LMP2-specific CTL response was induced in 323 out of 1×106mouse splenocytes. Conclusions High purity MSCs can be obtained using adherent culture;MSCs obtained by modified EBV LMP2（MSCs-LMP2）can induce LMP2 specific CTL response in the same strain of mice in vivo.

关 键 词：EB病毒潜伏膜蛋白2 间充质干细胞 细胞毒性T淋巴细胞 鼻咽癌 

分 类 号：R392[医药卫生—免疫学]