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Surface displaying of swine IgG1 Fc enhances baculovirus-vectored vaccine efficacy by facilitating viral complement escape and mammalian cell transduction

机译:猪IgG1 Fc的表面展示通过促进病毒补体逃逸和哺乳动物细胞转导来增强杆状病毒载体疫苗的效力

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摘要

Baculovirus-mediated gene transfer has been developed as a vaccine design strategy against a number of diseases without apparent viral replication. However, it has been hampered by complement-dependent inactivation, thus hindering the in vivo application of baculovirus. A variety of approaches have been exploited to bypass the complement system in the serum. In this study, we constructed and screened a series of baculovirus vectors displaying complement interfering factors, of which a baculovirus vector displaying swine IgG1 Fc (pFc) showed the highest complement antagonism (75.6%). Flow cytometry analysis of transduced cells demonstrated that the baculovirus display of pFc had a significant increase in transduction efficiency and transgene expression of reporter genes. On this basis, a VSV-G-pseudotyped with swine IgG1 Fc surface displayed baculovirus vector was developed to express the classical swine fever virus (CSFV) E2 gene. The translational enhancers Syn21 and P10UTR were incorporated to improve the antigen expression. The E2 gene was efficiently expressed in both insect and mammalian cells. Pigs immunized with this recombinant baculovirus developed high levels of E2-specific antibody, CSFV-specific neutralizing antibody and IFN-γ-secreting cellular immune responses. These results demonstrate that the strategy of surface-displaying swine IgG1 Fc has a great potential to improve the efficiency of baculovirus-vectored vaccine for CSFV and other swine pathogens.Electronic supplementary materialThe online version of this article (doi:10.1186/s13567-017-0434-5) contains supplementary material, which is available to authorized users.
机译:杆状病毒介导的基因转移已被开发为针对许多没有明显病毒复制的疾病的疫苗设计策略。然而,它已经被补体依赖性失活所阻碍,因此阻碍了杆状病毒的体内应用。已经开发出多种方法来绕过血清中的补体系统。在这项研究中,我们构建并筛选了一系列展示补体干扰因子的杆状病毒载体,其中展示猪IgG1 Fc(pFc)的杆状病毒载体表现出最高的补体拮抗作用(75.6%)。对转导细胞的流式细胞术分析表明,杆状病毒展示的pFc具有明显提高的报道基因转导效率和转基因表达。在此基础上,开发了以猪IgG1 Fc表面展示杆状病毒载体为假型的VSV-G型,以表达经典猪瘟病毒(CSFV)E2基因。整合翻译增强子Syn21和P10UTR以改善抗原表达。 E2基因在昆虫和哺乳动物细胞中均有效表达。用这种重组杆状病毒免疫的猪产生了高水平的E2特异性抗体,CSFV特异性中和抗体和分泌IFN-γ的细胞免疫应答。这些结果表明,表面展示猪IgG1 Fc的策略在提高杆状病毒载体疫苗对CSFV和其他猪病原体的效率方面具有巨大潜力。电子补充材料本文的在线版本(doi:10.1186 / s13567-017- 0434-5)包含补充材料,授权用户可以使用。

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