摘要
为探究猪圆环病毒2型(PCV2)优势T、B细胞表位对Cap蛋白免疫原性的影响,本研究将前期研究筛选到的PCV2 Rep和Cap蛋白中的T、B细胞表位,分别构建pET-rB-Cap、pET-rT-Cap和pET-rT-B-Cap重组质粒,并转化到大肠杆菌BL21中,经IPTG诱导表达后,采用SDS-PAGE和western blot检测3种重组蛋白:rB-Cap、rT-Cap和rT-B-Cap。结果显示,上述3种重组蛋白均高效表达且具有良好的反应原性。将70只4周龄~6周龄的BALB/c雌鼠随机分为7组,分别免疫不同的重组蛋白或者全病毒灭活疫苗,并对其产生的免疫原性进行检测。结果显示,各实验组小鼠的血清抗体水平均显著高于PBS组(p<0.01),其中rT-B-Cap组小鼠抗体水平显著高于PCV2全病毒灭活疫苗组和rCap组(p<0.01)。综上所述,PCV2 Cap蛋白同时串联T、B优势抗原表位时的免疫原性显著提高。本研究为PCV2多表位疫苗及基于Cap蛋白的亚单位疫苗的研究提供了新的途径。
The purpose of this study was to explore the effect of porcine circovirus type 2(PCV2) dominant T and B cell epitopes on the immunogenicity of Cap protein. The T and B cells epitopes in PCV2 Rep and Cap proteins identified in our previous research were used to construct the recombinant plasmids together this shortened Cap gene(pET-rB-Cap, pET-rT-Cap and pET-rT-B-Cap), respectively in this study. Then, the obtained plasmids were transformed into E.coli BL21. After induction by IPTG, three recombinant proteins(rB-Cap, rT-Cap, and rT-B-Cap) were identified by SDS-PAGE and western blot. The results showed that rB-Cap, rT-Cap, and rT-B-Cap were highly expressed and had good reactionogenicity. Seventy female BALB/c(4-6-weeks-old) mice randomly allocated into seven groups were immune with either different recombinant proteins(rCap, rB-Cap,rB-Cap, rT-Cap, rT-B-Cap and mixture proteins) or PCV2 inactivated vaccine. The immunogenicities of these were analyzed by testing the antibodies against PCV2 in mice. The results showed that there was significantly higher antibody levels in all the experimental groups than that in PBS control group(p<0.01). Meanwhile, the antibody levels was significantly higher in rT-B-Cap group than that in both PCV2 inactivated vaccine and rC ap groups(p<0.01). In conclusion, the immunogenicity of PCV2 Cap protein tandem with the T and B dominant epitopes was significantly improved in comparison with PCV2 inactivated vaccine. This study provides a new strategy for the study of PCV2 multi-epitope vaccine and Cap protein-based subunit vaccine.
作者
聂恺阳
吴云燕
易琳
吴祖雄
陈金顶
赵明秋
NIE Kai-yang;WU Yun-yan;YI Lin;WU Zu-xiong;CHEN Jin-ding;ZHAO Ming-qiu(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2019年第9期945-950,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家重点研发计划项目(2017YFD0500600、2016YFD0500700、2017YFD0501104)
国家自然科学基金(31472200)
广东省科技计划项目(2015B020230009)
广州市科技计划项目(201803020005)
