EB病毒潜伏膜蛋白2重组腺病毒的构建及其免疫效果的研究  

Effective Induction of Epstein-Barr Virus-Specific Cytotoxic T Lymphocytes in Mice by Latent Membrane Protein 2 Recombined Adenovirus

作　　者：姚家伟[1] 周玲[1] 王琦[1] 左建民[1] 曾毅[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《中国肿瘤》2003年第1期45-47,共3页China Cancer

基　　金：国家"973";"863"资助项目(G1998051201;2001AA217091)

摘　　要：为了进一步研究以EB病毒潜伏膜蛋白2穴LMP2)为靶抗原制备EB病毒相关肿瘤的治疗性疫苗,利用AdEasy系统构建了EBV鄄LMP2的重组腺病毒。PCR鉴定结果证实病毒DNA中含有目的基因的特异性片段;RT鄄PCR证明了外源基因在真核细胞中得以转录;免疫酶和Westernblot的结果也显示LMP2蛋白在真核细胞得到表达。将扩增后的病毒感染Hela细胞测定病毒滴度为1.5×109pfu/ml。将重组病毒以灌胃、肌肉注射和滴鼻的方式感染小鼠,经免疫荧光检测其特异性抗体,LDH法检测特异性CTL的杀伤作用,结果发现三种给药途径均可诱发小鼠针对LMP2的特异性体液免疫和细胞免疫,而用Ad5作为对照组的小鼠则没有引起相应的免疫反应。该研究将有益于进一步探索抗肿瘤特异性主动免疫作用在阻止肿瘤的生长、扩散或复发中的作用。对诱导机体产生特异性抗瘤活性有重要意义。In order to develop an EBV-LMP2vaccine,we constructed recombined adenovirus(rAd)bearing the full-length cDNA for LMP2driven by CMV.As time-consuming and tedious conventional procedure for construction of rAd,we chose AdEasy system to construct rAds.PCR and dot blot showed the LMP2gene integrated into the genome of rAds.LMP2specific mRNA was detected in infected293cells by RT-PCR.We obtained the LMP2recombinant adenovirus by Immunoenzyme method and Western blot assays.The titer of amplified rAds was1.5×10 9 Pfu/ml.Balb/c mice were immunized by the rAds with three routes.Preliminary results showed the rAds could elicit LMP2specific cellular and humoral immune responses.

关 键 词：EB病毒 潜伏膜蛋白2 重组腺病毒 构建 免疫效果 肿瘤 

分 类 号：R73－3[医药卫生—肿瘤]