小鼠B7-2基因转染EL4细胞作用的研究  

The study of mouse lymphoma cell EL4 transfected with mB7-2 gene

作　　者：秦雪梅[1] 马道新[1] 张庆英[2] 纪春岩[1] 徐从高[1] 

机构地区：[1]山东大学齐鲁医院血液科,济南250012 [2]青岛市第三人民医院,山东266041

出　　处：《中国免疫学杂志》2006年第7期600-603,共4页Chinese Journal of Immunology

摘　　要：目的:探讨小鼠免疫共刺激信号B7-2转化小鼠淋巴瘤细胞株EL4后,转化细胞介导的免疫作用。方法:(1)提取经LPS刺激后的小鼠脾细胞内的总RNA,以此为模板进行反转录PCR扩增,获得特异性的小鼠mB7-2 cDNA的产物并测序。将扩增的mB7-2 cDNA片段克隆到逆转录病毒表达载体pLXSN中,然后包装到PA317细胞中;(2)收集并浓缩PA317/mB7-2产生的病毒上清,感染EL4细胞,获得EL4/mB7-2;(3)体外进行混合淋巴细胞培养,检测EL4/mB7-2刺激小鼠脾脏淋巴细胞分泌IL-2情况。结果:成功得到了表达载体pLXSN-mB7-2和转基因的EL4/mB7-2。在体外,EL4/mB7-2刺激小鼠脾脏T淋巴细胞分泌的IL-2的量明显超过野生型EL-4细胞。结论:mB7-2基因转染EL-4细胞可活化T细胞分泌IL-2,为今后研究比较共刺激信号在肿瘤免疫、自身免疫病发病机制以及器官移植等方面的作用奠定基础。Objective：To assess the potential of mouse immune-costimulating signal B7-2 in inducing immune effect. Methods ： （ 1 ）The specific mB7-2cDNA fragment from LPS-stimulated mouse splenocytes was obtained by reverse transcription and polymerase chain reaction. The obtained fragment was inserted to pLXSN plasmid. Then the plasmid pLXSN-mB7-2 was packaged into PA317 cells;（2）The EL4/mB7-2 was obtained by infecting the EL4 by the concentrated virus particles produced by PA317/mB7-2; （3）In vitro, the secretion of IL-2 of EL4/mB7-2 stimulated-lymphocytes was detected by using mixed lymphocytes culture. Results： pLXSN-mBT-2 and transgenetic EL4/mB7-2 cells were obained successfully. IL-2 production in 24h in the supernatant stimulated by EL4/ mBT-2 was much higher than wild EL4 cells. Conclusion：EL4/mBT-2 can activated T cell. to produce IL-2. This research lay foundation for the research of the function of immune-costimulating signal in the tumor immunity and treatment, the mechanism of autoimmune disease and organ transplantation.

关 键 词：mB7-2分子 载体构建 EL4 生物活性 

分 类 号：R392[医药卫生—免疫学]