Zhou Xiaoyuan, Zhang Xinglang, Han Yahui, Jia Qiuhong, Gao Hongwei
Yellow River Fisheries Research Institute, Chinese Academy of Fishery Sciences, 2 Fenghui Road, South of Sanqiao, District of Weiyang, Xi'an 710086, China.
Viruses. 2017 Jul 25;9(8):195. doi: 10.3390/v9080195.
The Chinese giant salamander iridovirus (CGSIV), belonging to the genus in the family , is the causative agent of an emerging infectious disease causing high mortality of more than 90% and economic losses in Chinese giant salamanders in China. In this study, a recombinant baculovirus-based vaccine expressing the CGSIV major capsid protein (MCP) was developed and its protective immunity in Chinese giant salamanders was evaluated. The recombinant nucleopolyhedrosis virus (AcNPV), expressing CGSIV MCP, designated as AcNPV-MCP, was generated with the highest titers of 1 × 10⁸ plaque forming units/mL (PFU/mL) and confirmed by Western blot and indirect immunofluorescence (IIF) assays. Western blot analysis revealed that the expressed MCP reacted with mouse anti-MCP monoclonal antibodies at the band of about 53 kDa. The results of IIF indicated that the MCP was expressed in the infected 9 (Sf9) cells with the recombinant baculovirus, and the Chinese giant salamander muscle cells also transduced with the AcNPV-MCP. Immunization with the recombinant baculovirus of AcNPV-MCP elicited robust specific humoral immune responses detected by ELISA and neutralization assays and potent cellular immune responses in Chinese giant salamanders. Importantly, the effective immunization conferred highly protective immunity for Chinese giant salamanders against CGSIV challenge and produced a relative percent of survival rate of 84%. Thus, the recombinant baculovirus expressing CGSIV MCP can induce significant immune responses involving both humoral and cell-mediated immunity in Chinese giant salamanders and might represent a potential baculovirus based vaccine candidate for Chinese giant salamanders against CGSIV.
中国大鲵虹彩病毒(CGSIV)属于 科 属,是一种新兴传染病的病原体,在中国导致中国大鲵死亡率超过90%并造成经济损失。在本研究中,开发了一种表达CGSIV主要衣壳蛋白(MCP)的重组杆状病毒疫苗,并评估了其在中国大鲵中的保护性免疫。构建了表达CGSIV MCP的重组苜蓿银纹夜蛾核型多角体病毒(AcNPV),命名为AcNPV-MCP,其最高滴度为1×10⁸ 空斑形成单位/毫升(PFU/毫升),并通过蛋白质免疫印迹法和间接免疫荧光(IIF)分析进行了确认。蛋白质免疫印迹分析显示,表达的MCP在约53 kDa的条带处与小鼠抗MCP单克隆抗体发生反应。IIF结果表明,MCP在感染重组杆状病毒的草地贪夜蛾细胞系9(Sf9)中表达,中国大鲵肌肉细胞也被AcNPV-MCP转导。用AcNPV-MCP重组杆状病毒免疫可引发通过ELISA和中和试验检测到的强烈特异性体液免疫反应以及中国大鲵中的有效细胞免疫反应。重要的是,有效免疫为中国大鲵提供了针对CGSIV攻击的高度保护性免疫,产生了84%的相对存活率。因此,表达CGSIV MCP的重组杆状病毒可在中国大鲵中诱导涉及体液免疫和细胞介导免疫的显著免疫反应,可能代表一种针对中国大鲵抵抗CGSIV的潜在杆状病毒疫苗候选物。