摘要
为评价分子佐剂细胞毒性T淋巴细胞相关抗原4(CTLA-4)对IBDV VP2质粒DNA的免疫增强作用,试验采用重组质粒载体表达重组蛋白mLTA-VP2-CTLA-4和mLTA-CTLA-4,家兔毒性试验确定其安全性,后选择10日龄非免疫健康鸡进行随机分组试验,设不同剂量mLTA-CTLA-4加IBD活疫苗免疫组、不同剂量mLTAVP2-CTLA-4免疫组、IBD活疫苗免疫组及空白对照组,免疫后用ELISA法定期检测鸡血清抗体IgG及小肠黏膜抗体IgA效价;鸡在加强免疫后用IBDV野生毒株攻击,连续观察2周并计算保护率。结果显示,mLTA-VP2-CTLA-4免疫组、mLTA-CTLA-4加活疫苗共同免疫组产生的IgG和IgA抗体水平与活疫苗免疫组无明显差别,mLTA-CTLA-4加活疫苗共同免疫组产生的IgG和IgA抗体水平略高于mLTA-VP2-CTLA-4免疫组,疫苗对鸡的保护率为100%。结果表明,构建的IBDV分子佐剂DNA疫苗载体能表达无毒性的重组蛋白质,并能产生很好的免疫保护率,为进一步研究IBD亚单位疫苗奠定基础。
The study was conducted to evaluate the immune enhancement of molecular adjuvant CTLA-4on IBDV VP2 plasmid DNA.Fusion protein mLTA-CTLA-4and mLTA-VP2-CTLA-4were expressed and purified based on vector pET-mLTA-CTLA-4and pET-mLTA-VP2-CTLA-4.Protein toxicity tests were carried out on rabbits.10-day-old chickens were randomly divided into four groups,including different doses of mLTA-CTLA-4plus IBD vaccine groups,different doses of mLTA-VP2-CTLA-4groups,IBD live vaccine groups and control groups.Serum and mucosal samples were regularly collected for neutralization titer of IgG and IgA.After booster immunization,chickens were attacked by wild IBDV strain,and two weeks later protection rate were calculated.The results showed that levels of IgG and IgA in mLTA-VP2-CTLA-4groups,mLTACTLA-4with IBDV vaccine groups and conventional IBDV vaccine groups had no significant difference.IgG and IgA levels in mLTA-CTLA-4with IBDV vaccine groups were slightly higherthan that of mLTA-VP2-CTLA-4groups.The protection rate was 100%.The results indicate that constructed IBDV molecular adjuvant DNA vaccine vector can express non-toxic recombinant protein,which can produce a good immune efficiency in chickens.It lays a foundation for further study on IBD subunit vaccine.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2015年第5期824-829,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
江苏省博士后科研资助计划(1401077B)
博士后日常资助项目
江苏农牧科技学院"凤凰人才工程"项目
江苏农牧科技职业学院重点支持项目(NSFZD1405
NSFZD1305)
扬州朝天歌农牧科技有限公司横向合作课题(00010114012)
江苏省兽用生物制药高技术研究重点实验室开放课题(JSKLKF1403)
