猪瘟病毒E2蛋白靶向化树突状细胞疫苗诱导的BALB/c小鼠体内/外免疫应答的研究  

The immune response of classic a swine fever virus E2 protein targeting to dendritic cell vaccine in vivo/vitro of BALB/c mice

作　　者：董炳梅 谢金文[2] 魏凤[2] 孙全文[3] 王爱华[3] 张春玲 沈志强[1,2] 王金良[2] 

机构地区：[1]山东绿都生物科技有限公司,山东滨州256600 [2]山东省滨州畜牧兽医研究院,山东滨州256600 [3]河北北方学院,河北张家口075051

出　　处：《中国预防兽医学报》2014年第4期305-309,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：山东省自然科学基金项目(ZR2010CQ012);河北北方学院重大项目(zd201307)

摘　　要：为确定猪瘟病毒(CSFV)糖基化E2蛋白(Gly-E2)的免疫效力,本研究首先利用体外糖基化修饰技术,在对E2蛋白(E2)进行N-糖基化修饰的基础上,分别进行Gly-E2、DCs与T细胞体外共培养并接种BALB/c小鼠。通过ELISA方法对细胞上清液及BALB/c小鼠血清中IFN-γ、IL-4及IgG含量进行测定,结合体外试验中CTL特异性杀伤率的检测,综合评价该疫苗的免疫效果。结果显示,CSFV E2的糖基化修饰显著增强了DCs的抗原提呈功能,可以同时促进Th1与Th2免疫应答,但主要以Th2体液免疫应答为主,刺激机体产生了快而持久的抗体保护,并且具有良好的免疫记忆,其免疫效果显著优于CSFV与E2;同时,随着Gly-E2浓度的提高,效应细胞的杀伤率均明显升高,表明Gly-E2可以更有效的刺激效应细胞的杀伤作用。以上结果表明,Gly-E2对提高CSFV的免疫反应具有良好的效果,作为新型疫苗开发的优选抗原,Gly-E2具有很深的开发潜力。In order to evaluate the immune efficacy of glycosylated E2 protein （Gly-E2） of classic swine fever virus （CSFV）, the Gly-E2, dendritic cells （DCs） and T cells were co-cultured and the BALB/c mice were inoculated with the Gly-E2. Consequently, the IL-4, IFN-γand IgG levels in cell culture supernatant and serum of BALB/c mice were detected by ELISA, and the rate of CTL specific killing was evaluated in vitro. The results show that the glycosylation of E2 enhanced antigen presentation of dendritic cells significantly, and promoted the Thl and Th2 immune response at the same time. In addition, the Th2 humoral immune response was more significant. The Gly-E2 sitimulated the body to produce quick and more durable antibody and had a better immune memory than that of CSFV and E2. Furthermore, the rate of killing effector cells significantly increased in a dose depend manner with the Gly-E2. These results proved that the Gly-E2 possessed a good preventive effect to classical swine fever. Therefore, the Gly-E2 has a potential for development of new vaccines.

关 键 词：猪瘟病毒 E2蛋白 树突状细胞疫苗 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]