首页> 外文期刊>Journal of Virological Methods >Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine circovirus type 2 in mice.
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Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine circovirus type 2 in mice.

机译:表达2型猪圆环病毒衣壳蛋白的重组假型杆状病毒的构建及其免疫原性。

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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.
机译:杆状病毒最近作为哺乳动物细胞的新颖且有吸引力的基因递送载体而出现。已知2型猪圆环病毒(PCV2)与断奶后多系统消耗综合症(PMWS)有关,这是一种正在出现的猪病,造成巨大的经济损失。在这项研究中,假性与水泡性口腔炎病毒糖蛋白(VSV-G)杆状病毒被用作表达衣壳蛋白(Cap)的载体,衣壳蛋白是PCV2最重要的免疫原,在巨细胞病毒即刻早期(CMV-IE)增强子的转录控制下/发起人。 Western印迹和流式细胞仪分析表明,所得重组杆状病毒(BV-G-ORF2)在哺乳动物细胞中有效转导并表达了Cap蛋白。用1x10(8)或1x10(9)斑块形成单位(PFU)/小鼠BV-G-ORF2直接接种疫苗后,可以在小鼠中诱导出重要的PCV2特异性ELISA抗体,中和抗体以及细胞免疫应答。即使以1x10(8)PFU /小鼠的剂量,BV-G-ORF2也比编码Cap蛋白的DNA疫苗具有更好的免疫原性。综上所述,BV-G-ORF2的增强的免疫原性以及假型杆状病毒的独特优势,包括易于操作,简单的按比例放大,无毒性以及宿主中不存在针对杆状病毒的抗体,表明该假型杆状病毒介导的基因传递可以用作开发新一代抗PCV2感染疫苗的替代策略。

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