猪圆环病毒2型ORF2/猪白介素2嵌合表达质粒在猪体内诱导的保护性免疫反应  

Protective immunity reaction induced by porcine circovirus type 2(PCV2) ORF2/porcine interleukin-2(PoIL-2) expression plasmid in vivo

作　　者：闫若潜[1] 王东方[1] 吴志明[1] 李桂喜 刘淑敏[3] 刘梅芬[1] 赵明军[1] 王淑娟[1] 赵雪丽[1] 

机构地区：[1]河南省动物疫病预防控制中心,河南郑州450008 [2]河南商丘市动物疫病预防控制中心,河南商丘476000 [3]郑州市中心医院消毒供应中心,河南郑州450007

出　　处：《中国兽医学报》2014年第2期199-206,共8页Chinese Journal of Veterinary Science

基　　金：河南省科技创新人才计划资助项目(豫科人组[2011]1号)

摘　　要：为探讨猪圆环病毒2型(PCV2)ORF2/猪白介素-2(PoIL-2)嵌合重组表达质粒(rpcDNA3.1/PCV2-linker-PoIL-2)在猪体内的免疫效果和免疫保护效果进而研制高效PCV2核酸疫苗,将35只10日龄健康仔猪平均分成7组分别以rpcDNA3.1/PCV2-linker-PoIL-2重组表达质粒或PCV2ORF2基因原核表达的rCap蛋白进行免疫及免疫保护试验。共进行4次肌肉注射免疫,每次间隔2周;于第4次免疫后3周通过口腔和鼻腔途径感染PCV2细胞强毒。分别于第4次免疫后和攻毒后不同时间通过检测免疫猪血清抗体水平和外周血T淋巴细胞增殖活性、辅助性T细胞(Th)和细胞毒性T细胞(Tc)亚群的百分含量、排毒率和病毒血症阳性维持时间等指标以评价其免疫和免疫保护效果。结果表明,各免疫组猪均产生了抗PCV2特异性ELISA免疫抗体,但rCap蛋白免疫组猪抗体水平较低;rpcDNA3.1/PCV2-linker-PoIL-2质粒对猪体的免疫和免疫保护效果显著优于rpcDNA3.1/ORF2质粒;在rpcDNA3.1/PCV2-linker-PoIL-2质粒或rpcDNA3.1/ORF2质粒中添加rCap蛋白对重组质粒免疫及免疫保护效果无明显影响。因此,选取rpcDNA3.1/PCV2-linker-PoIL-2质粒为下一步研制PCV2核酸疫苗的主要成分。In order to explore a good candidate recombinant expression plasmid for porcine circovir us type 2 （PCV2） DNA vaccines, the immune effects and immunoprotection effects of the chimeric recombinant expression plasmid of rpcDNA3.1/PCV2-1inker-PoIL-2 which had been constructed by PCV20RF2 / porcine interleukin-2 （PoIL-2） in our previous study were evaluated. Thirty-five 10-day-old healthy piglets were equally divided into 7 groups which were immunized by rpcDNA 3.1/PCV2-1inker-PoIL-2 plasmid or rCap protein derived by recombinant prokaryotic expression by ORF2 of PCV2,respectively. All piglets were injected intramuscularly four times at 2-week in tervals. At 3 weeks after the fourth immunization, all pigs were challenged with PCV2 cells viru lent strain by intraoral and intranasal ways. After the fourth immunization and the different inter val times after challenge by PCV2 cells virulent strain,the immune effects and immunoprotection effects were evaluated by detection the indicators of swine immune serum antibody levels, theproliferation activity of peripheral blood T lymphocytes, the percentage contents of the T helper cells （Th） and cytotoxic T cells （Tc） ,the percentage of viral excretion and maintain time of virae mia,respectively. The results showed that all the immunized pigs produced the specific antibodies against PCV2, whereas the antibody levels of the pigs only immunized by rCap protein were lower. The immune effects and immunoprotection effects of groups immunized with rpcDNA3.1/PCV2- linker-PolL-2 plasmid were significantly better than the group immunized with rpcDNA3.1/ORF2 plasmid, and there were no obvious enhancing effects on immunization and immunoprotection when added rCap protein to the rpcDNA3.1/PCV2-1inker-PolL-2 or rpcDNA3.1/ORF2 plasmids. Therefore,the rpcDNA3.1/PCV2-1inker-PolL 2 plasmid will be selected as the main candidate in gredient of PCV2 DNA vaccine for the further study.

关 键 词：猪圆环病毒2型 猪白细胞介素2 rpcDNA3 1 PCV2-linker—PoIL-2重组表达质粒 免疫保护 

分 类 号：S852.65[农业科学—基础兽医学]