表达猪白介素2基因重组腺病毒的构建及其免疫增强作用  

Construction and Immunoenhancement of a Recombinant Adenoviruses Encoding Porcine Interleukin 2

作　　者：何雷[1] 张彦明[1] 向华[1] 唐青海[1] 徐彦召[1] 杨小云[1] 王静[1] 代晨[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农业学报》2010年第11期1-7,共7页Acta Agriculturae Boreali-occidentalis Sinica

基　　金："863"重大项目"家畜重要病毒病基因工程疫苗研究和创制"(2006AA10A204)

摘　　要：以猪白介素2基因(pIL-2)为研究对象构建表达pIL-2的重组腺病毒,并研究其作为细胞因子佐剂的免疫增强作用。用RT-PCR的方法扩增pIL-2基因,将pIL-2基因克隆到腺病毒穿梭载体AdTrack中,构建出重组腺病毒穿梭质粒AdTrack-pIL-2,然后在大肠杆菌BJ5183内和骨架载体AdEasy同源重组,获得重组腺病毒骨架载体AdEasy-pIL-2,转染HEK293细胞,包装出重组腺病毒Ad-pIL-2,并通过兔体交互试验来检测Ad-pIL-2的免疫增强作用。经PCR、RT-PCR和Western blot检测,成功构建了Ad-pIL-2,病毒滴度可以达到108.25pfu/mL,并通过兔体交互免疫试验证明Ad-pIL-2可以提高兔的抗体水平。用腺病毒构建的Ad-pIL-2在兔体上具有免疫增强作用,为下一步的猪体试验研究和细胞因子佐剂的开发奠定基础。The recombinant adenovirus carrying porcine interleukin 2（pIL-2） was constructed and the immunologic enhancement of it was studied as a cytokine adjuvant.The pIL-2 gene fragment was amplified by RT-PCR and was inserted into adenovirus shuttle vector AdTrack-CMV to produce the AdTrack-pIL-2.Linearized by Pme I,it was transferred into E.coli BJ5183 cells where AdEasy-pIL-2 was generated.The recombinant adenovirus Ad-pIL-2 was propagated after transfecting AdEasy-pIL-2 into HEK293 cells,and the immunoenhancement of it was evaluated on rabbits then.pAd-pIL-2 could carry and express pIL-2 protein by identification of PCR,RT-PCR and Western-blot.The titer of it was 10^8.25 pfu/mL.It could improve the IgG Level of rabbits compared to the group without being inoculated.Recombinant adenovirus vector Ad-pIL-2 was constructed successfully and the immune enhancing effect of it was evaluated on rabbits,which may lay a foundation for the further study of the effect on pigs as a cytokine adjuvant.

关 键 词：猪白介素2 重组腺病毒 细胞因子 佐剂 

分 类 号：S852.5[农业科学—基础兽医学]