摘要
为评价CpG基序对新城疫病毒(NDV)DNA免疫效果的影响,本研究将NDVF48E9株的F基因分别克隆至真核表达载体pVAX1和含有3个CpG基序的载体pVAX1-CpG中,分别命名为pV-F和pVC-F,体外表达检测结果显示,pV-F和pVC-F外源基因表达量无明显差别。将pVAX1、pVAX1-CpG、pV-F和pVC-F经腿部肌肉多点注射3周龄SPF鸡(200μg/只),2周后以相同的剂量加强免疫,二免后2周通过滴鼻以103EID50NDV的F48E9强毒株进行攻毒。抗体检测结果表明,免疫组的抗体水平与对照组相比差异显著(p<0.01),pVC-F免疫组的抗体水平高于pV-F免疫组;淋巴细胞检测结果表明,pV-F和pVC-F免疫组与对照组显著差异(p<0.01);攻毒保护率结果显示,pVC-F和pV-F组的保护率分别为60%和50%,对照组为0。以上结果表明,CpG基序对机体细胞免疫反应有一定的增强作用。
To investigate the effects of CpG oligonucleotides on immune efficacy of DNA immunization,the F gene of NDV F48E9 strain was cloned to vector pVAX1 and pVAX1-CpG（containing three CpG sequences）,respectively,to obtain recombinant plasmids pV-F and pVC-F.The recombinant plasmids were transfected into 293T cells,respectively,and F gene expression were detected by indirect immunofluorescence and western blot.The results showed that no obvious difference in F gene expression was detected from pV-F and pVC-F.SPF chickens were injected with plasmids pV-F and pVC-F at 200 μg for each chicken and boosted with the same dose three weeks later.The antibody level of immunization groups was significantly higher（p0.01） than that of the controls,the percentage of CD4＋ and CD8＋ T lymphocytes in the peripheral blood of the inoculated groups was also significantly higher（p0.01）.The injected chickens showed 60 %（PVC-F） and 50 %（PV-F） protection after challenge with 1000 EID50 NDV F48E9 strain three weeks after the second immunization.Somehow,the CpG sequence showed immune enhancement to the DNA immunization.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第7期554-558,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
兽医生物技术国家重点实验室基本科研业务费(NKLVBP200803)
