EB病毒潜伏膜蛋白2DNA疫苗的构建及其初步免疫效果观察  

Induction of Epstein-Barr virus-specific cytotoxic T lymphocytes by using latent membrane protein 2 DNA vaccine

作　　者：朱伟严[1] 周玲[1] 王琦[1] 姚家伟[1] 曾毅[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052

出　　处：《中华微生物学和免疫学杂志》2002年第2期185-190,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家" 973"资助项目;" 86 3"资助项目 (G19980 5 12 0 1;2 0 0 1AA2 170 91)

摘　　要：目的　以EB病毒潜伏膜蛋白 2 (latentmembraneprotein 2 ,LMP2 )为靶基因 ,构建EBV LMP2的侯选DNA疫苗 ;并初步探讨其在小鼠体内诱导特异性细胞毒方面的作用 ,为研制鼻咽癌 (na sopharyngealcarcinoma ,NPC)等EB病毒相关肿瘤的治疗性疫苗提供有益资料。方法　将EB病毒LMP2全cDNA片段克隆至含CMV早期启动子的真核表达载体pcDNAⅢ上 ,构建CMV启动子EB病毒LMP2DNA疫苗。在体外将重组质粒转染COS细胞 ,以RT PCR和间接免疫荧光法检测重组质粒在转染的COS细胞中的转录和表达。采用 5 0 μg、10 0 μg、2 0 0 μg 3种质粒剂量进行初步的小鼠免疫试验。结果　重组质粒可在真核细胞中有效地转录和表达LMP2。免疫接种小鼠后可诱发机体产生针对LMP2蛋白的特异性体液免疫和细胞免疫。 5 0 μg、10 0 μg、2 0 0 μg 3种免疫剂量产生的抗体水平差异不大。在免疫接种 6周后 ,重组质粒免疫的小鼠产生针对LMP2的特异性CTL均明显高于空载体免疫小鼠。 3种剂量的CTL结果显示 :在 10 0 μg、2 0 0 μg免疫组 ,小鼠诱导产生的CTL水平要略高于 5 0 μg免疫组。结论　EB病毒重组质粒LMP2免疫小鼠可以诱发小鼠产生特异的体液和细胞免疫应答。Objective To develop an EBV-LMP2 DNA vaccine, we have constructed a plasmid DNA encoding the full-length cDNA for LMP2 driven by CMV. Methods The recombined plasmids expressed LMP2 well in COS7 cells and after immunization with it the BALB/c mice induced both cellular and humoral immune responses. To charaterize the effect of DNA vaccine dosage, three groups of mice were immunized with 50μg, 100μg, 200μg doses of pcDNAⅢ-LMP2 respectively. Results Three groups showed no difference in inducing humoral immune response. But in inducing cellular immune response, of 50μg were less effective than doses 100μg, 200μg. Conclusion Our results recommend LMP2 vaccine in future clinical studies. In EBV-seropositive individuals, if LMP2 protein can be presented in endogenously process way to induce specific CTL responses in EBV-IgA/VCA-seropositive individuals, it can protect host against cancer development and establish the CTL-based tumor therapy for NPC. The present work may be useful to the development of novel vaccines of EBV associated tumors.

关 键 词：EB病毒 潜伏膜蛋白2 特异性细胞毒作用 核酸疫苗 DNA疫苗 免疫效果观察 

分 类 号：R392[医药卫生—免疫学]