携带外源表位的嵌合重组2型猪圆环病毒病毒样颗粒作为抗原载体

Virus-like particles of chimeric recombinant porcine circovirus type 2 as antigen vehicle carrying foreign epitopes.

作者信息

Zhang Huawei, Qian Ping, Liu Lifeng, Qian Suhong, Chen Huanchun, Li Xiangmin

机构信息

State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.

出版信息

Viruses. 2014 Dec 5;6(12):4839-55. doi: 10.3390/v6124839.

Virus-like particles (VLPs) of chimeric porcine circovirus type 2 (PCV2) were generated by replacing the nuclear localization signal (NLS; at 1-39 aa) of PCV2 capsid protein (Cap) with classical swine fever virus (CSFV) T-cell epitope (1446-1460 aa), CSFV B-cell epitope (693-716 aa) and CSFV T-cell epitope conjugated with B-cell epitope. The recombinant proteins were expressed using the baculovirus expression system and detected by immunoblotting and indirect immunofluorescence assay. The abilities to form PCV2 VLPs were confirmed by transmission electron microscopy. Immunogenicities of the three recombinant proteins were evaluated in mice. Our Results indicated that Cap protein NLS deletion or substitution with CSFV epitopes did not affect the VLPs assembly. Three chimeric Cap proteins could form VLPs and induce efficient humoral and cellular immunity against PCV2 and CSFV in mice. Results show that PCV2 VLPs can be used as an efficient antigen carrier for delivery of foreign epitopes, and a potential novel vaccine.

通过用经典猪瘟病毒（CSFV）T细胞表位（第1446 - 1460位氨基酸）、CSFV B细胞表位（第693 - 716位氨基酸）以及与B细胞表位偶联的CSFV T细胞表位替换猪圆环病毒2型（PCV2）衣壳蛋白（Cap）的核定位信号（NLS；第1 - 39位氨基酸），产生了嵌合型PCV2病毒样颗粒（VLPs）。使用杆状病毒表达系统表达重组蛋白，并通过免疫印迹和间接免疫荧光测定法进行检测。通过透射电子显微镜确认形成PCV2 VLPs的能力。在小鼠中评估了这三种重组蛋白的免疫原性。我们的结果表明，Cap蛋白NLS缺失或被CSFV表位取代不影响VLPs组装。三种嵌合Cap蛋白可形成VLPs，并在小鼠中诱导针对PCV2和CSFV的高效体液免疫和细胞免疫。结果表明，PCV2 VLPs可作为递送外源表位的有效抗原载体，以及一种潜在的新型疫苗。