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利用痘病毒作为载体优化 ASFV 蛋白的表达用于亚单位疫苗的开发。

Optimization in the expression of ASFV proteins for the development of subunit vaccines using poxviruses as delivery vectors.

机构信息

Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, 53706, USA.

Plum Island Animal Disease Center, Agricultural Research Service (ARS), Greenport, NY, 11944, USA.

出版信息

Sci Rep. 2021 Dec 6;11(1):23476. doi: 10.1038/s41598-021-02949-x.

Abstract

African swine fever virus (ASFV) causes a highly contagious hemorrhagic disease that affects domestic pig and Eurasian wild boar populations. To date, no safe and efficacious treatment or vaccine against ASF is available. Nevertheless, there are several reports of protection elicited by experimental vaccines based on live attenuated ASFV and some levels of protection and reduced viremia in other approaches such as DNA, adenovirus, baculovirus, and vaccinia-based vaccines. Current ASF subunit vaccine research focuses mainly on delivering protective antigens and antigen discovery within the ASFV genome. However, due to the complex nature of ASFV, expression vectors need to be optimized to improve their immunogenicity. Therefore, in the present study, we constructed several recombinant MVA vectors to evaluate the efficiency of different promoters and secretory signal sequences in the expression and immunogenicity of the p30 protein from ASFV. Overall, the natural poxvirus PrMVA13.5L promoter induced high levels of both p30 mRNA and specific anti-p30 antibodies in mice. In contrast, the synthetic PrS5E promoter and the S E/L promoter linked to a secretory signal showed lower mRNA levels and antibodies. These findings indicate that promoter selection may be as crucial as the antigen used to develop ASFV subunit vaccines using MVA as the delivery vector.

摘要

非洲猪瘟病毒(ASFV)引起一种高度传染性的出血性疾病,影响家猪和欧亚野猪种群。迄今为止,尚无针对 ASF 的安全有效的治疗或疫苗。然而,有几篇报道称,基于减毒活 ASFV 的实验疫苗可引起保护作用,并且在其他方法(如 DNA、腺病毒、杆状病毒和痘苗病毒疫苗)中可引起一定程度的保护和降低病毒血症。目前,ASF 亚单位疫苗的研究主要集中在 ASFV 基因组内递送保护性抗原和抗原发现上。然而,由于 ASFV 的复杂性,需要优化表达载体以提高其免疫原性。因此,在本研究中,我们构建了几种重组 MVA 载体,以评估不同启动子和分泌信号序列在 ASFV p30 蛋白表达和免疫原性中的效率。总的来说,天然痘病毒 PrMVA13.5L 启动子在小鼠中诱导了高水平的 p30 mRNA 和特异性抗 p30 抗体。相比之下,合成的 PrS5E 启动子和连接分泌信号的 S E/L 启动子显示出较低的 mRNA 水平和抗体。这些发现表明,启动子的选择可能与抗原一样重要,对于使用 MVA 作为递送载体开发 ASFV 亚单位疫苗至关重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d0ac/8648923/2f2b55099604/41598_2021_2949_Fig1_HTML.jpg

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