Fan Huiying, Pan Yongfei, Fang Liurong, Wang Dang, Wang Shengping, Jiang Yunbo, Chen Huanchun, Xiao Shaobo
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, 1 Shi-zi-shan Street, Wuhan 430070, China.
J Virol Methods. 2008 Jun;150(1-2):21-6. doi: 10.1016/j.jviromet.2008.02.011. Epub 2008 Apr 3.
Baculovirus has emerged recently as a novel and attractive gene delivery vehicle for mammalian cells. Porcine circovirus type 2 (PCV2) is known to be associated with post-weaning multisystemic wasting syndrome (PMWS), an emerging swine disease which results in tremendous economic losses. In this study, baculovirus pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G) was used as a vector to express capsid (Cap) protein, the most important immunogen of PCV2, under the transcriptional control of cytomegalovirus immediate early (CMV-IE) enhancer/promoter. The resultant recombinant baculovirus (BV-G-ORF2) efficiently transduced and expressed the Cap protein in mammalian cells, as demonstrated by Western blot and flow cytometric analyses. After direct vaccination with 1x10(8) or 1x10(9)plaque forming units (PFU)/mouse of BV-G-ORF2, significant PCV2-specific ELISA antibodies, neutralizing antibodies, as well as cellular immune responses could be induced in mice. BV-G-ORF2 exhibited better immunogenicity than a DNA vaccine encoding the Cap protein, even at a dose of 1x10(8)PFU/mouse. Taken together, the improved immunogenicity of BV-G-ORF2, together with the unique advantages of pseudotype baculovirus, including easy manipulation, simple scale-up, lack of toxicity, and no pre-existing antibody against baculovirus in the hosts, indicate that pseudotype baculovirus-mediated gene delivery can be utilized as an alternative strategy to develop a new generation of vaccines against PCV2 infection.
杆状病毒最近已成为一种用于哺乳动物细胞的新型且有吸引力的基因递送载体。已知猪圆环病毒2型(PCV2)与断奶后多系统消瘦综合征(PMWS)有关,PMWS是一种新出现的猪病,会导致巨大的经济损失。在本研究中,用泡状口炎病毒糖蛋白(VSV-G)假型化的杆状病毒被用作载体,在巨细胞病毒立即早期(CMV-IE)增强子/启动子的转录控制下表达PCV2最重要的免疫原衣壳(Cap)蛋白。如蛋白质免疫印迹和流式细胞术分析所示,所得重组杆状病毒(BV-G-ORF2)在哺乳动物细胞中有效地转导并表达了Cap蛋白。用1×10⁸或1×10⁹蚀斑形成单位(PFU)/小鼠的BV-G-ORF2直接免疫小鼠后,可在小鼠体内诱导出显著的PCV2特异性ELISA抗体、中和抗体以及细胞免疫反应。即使在剂量为1×10⁸PFU/小鼠时,BV-G-ORF2也比编码Cap蛋白的DNA疫苗表现出更好的免疫原性。综上所述,BV-G-ORF2免疫原性的提高,以及假型杆状病毒的独特优势,包括易于操作、简单放大、无毒性以及宿主中不存在针对杆状病毒的预先存在的抗体,表明假型杆状病毒介导的基因递送可作为开发新一代抗PCV2感染疫苗的替代策略。