表达EB病毒潜伏膜蛋白2A(LMP2A)的小鼠移植瘤模型的建立

Establishment of the Animal Model Implanted With Tumor Expressing EB Virus Latent Membrane Protein 2A(LMP2A)

目的:建立表达EB病毒LMP2A小鼠移植瘤模型,为树突状细胞治疗EB病毒相关肿瘤的动物试验奠定基础。方法:将EB病毒LMP2A基因克隆至逆转录质粒pLXSN中,重组质粒用脂质体Clonfectin转入BALB/c小鼠来源的骨髓瘤细胞SP2/0,经G418筛选获得阳性克隆,亚克隆得到EB病毒LMP2A表达株并经PCR、RT-PCR、IFA、流式细胞仪鉴定。腹股沟皮下接种该细胞入BALB/c小鼠得到移植瘤模型,两周后切除肿瘤组织并作苏木精-伊红(H-E)染色切片。结果:构建出含EB病毒LMP2A基因的重组逆转录质粒,获得表达EB病毒LMP2A的SP2/0细胞,该细胞在BALB/c小鼠可生长成肿块。结论:成功地构建表达EB病毒潜伏膜蛋白2A(LMP2A)的小鼠移植瘤模型。

Objective: To establish an animal model implanted with EB virus LMP2A expressing tumor for the evaluation of dendritic cells (DCs) based immunotherapy of EB virus associated tumors in vivo. Methods: The EB virus LMP2A was cloned into retrovirus vector pLXSN. The recombinant vector was transfected into BALB/c mice derived SP2/0 cells by Clonfectin and then cells were screened by G418 to get positive clones and amplified. Positive cells were identified by means of PCR and, RT-PCR, IFA, FACS test. Those cells were injected into BABL/c mice to establish tumor animal model. The tumor was removed after 2 weeks and subjected to histological examination (H-E stain). Results: The retrovirus vector containing LMP2A gene was constructed. The SP2/0 cells expressing LMP2A were established. Those LMP2A-expressing SP2/0 cells can become tumors in BALB/c mice. Conclusion: The animal model implanted with. EB virus LMP2A expressing tumor was established.