College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.
BMC Vet Res. 2020 Feb 7;16(1):45. doi: 10.1186/s12917-020-2253-6.
Porcine circovirus type 2 (PCV2) is an economically important pathogen in the swine industry worldwide. Vaccination remains the principal tool to control PCV2-associated diseases (PCVADs). Current vaccines do not eliminate viral shedding in the environment. To enhance the efficacy of PCV2 vaccines, recombinant virus-like particles (VLPs) of PCV2 were generated by fusing a truncated form of flagellin FliC (TFlg: 85-111aa) with the PCV2 capsid protein (Cap).
The recombinant proteins were expressed in Escherichia coli and detected using Western blotting. The abilities of the recombinant proteins to assemble into VLPs were observed under transmission electron microscopy (TEM). The protective immune responses of recombinant VLPs were further evaluated by immunization of mice. The results showed that insertion of TFlg into C terminal of the Cap protein did not affect the formation of VLPs and boosted both humoral and cellular immune responses in mice. After a challenge with PCV2, in the Cap-TFlg vaccinated group, viremia was milder and viral loads were lower as compared with those in the Cap vaccinated group.
These results suggest that recombinant VLPs of PCV2 containing a TFlg adjuvant can be used as a promising PCV2 vaccine candidate.
猪圆环病毒 2 型(PCV2)是全球养猪业中一种具有重要经济意义的病原体。疫苗接种仍然是控制与 PCV2 相关疾病(PCVADs)的主要手段。目前的疫苗并不能消除环境中的病毒脱落。为了提高 PCV2 疫苗的效力,通过将 flagellin FliC 的截短形式(TFlg:85-111aa)与 PCV2 衣壳蛋白(Cap)融合,生成了重组病毒样颗粒(VLPs)。
重组蛋白在大肠杆菌中表达,并通过 Western blot 检测。使用透射电子显微镜(TEM)观察重组蛋白组装成 VLPs 的能力。通过免疫小鼠进一步评估了重组 VLPs 的保护免疫反应。结果表明,将 TFlg 插入 Cap 蛋白的 C 末端不会影响 VLPs 的形成,并增强了小鼠的体液和细胞免疫反应。在受到 PCV2 挑战后,与 Cap 疫苗接种组相比,Cap-TFlg 疫苗接种组的病毒血症较轻,病毒载量较低。
这些结果表明,含有 TFlg 佐剂的 PCV2 重组 VLPs 可用作有前途的 PCV2 疫苗候选物。