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自组装貉细小病毒 VP2 蛋白赋予貉对 RDPV 疾病的免疫力:体外和体内研究。

Self-assembled raccoon dog parvovirus VP2 protein confers immunity against RDPV disease in raccoon dogs: in vitro and in vivo studies.

机构信息

School of Life Sciences, Changchun University of Technology, Changchun, 130012, China.

Institute of Special Animal and Plant Sciences of CAAS, Changchun, 130012, China.

出版信息

Virol J. 2021 Apr 15;18(1):79. doi: 10.1186/s12985-021-01549-5.

Abstract

BACKGROUND

Raccoon dog parvovirus (RDPV) causes acute infectious diseases in raccoon dogs and may cause death in severe cases. The current treatment strategy relies on the extensive usage of classical inactivated vaccine which is marred by large doses, short immunization cycles and safety concerns.

METHODS

The present study aimed at optimization of RDPV VP2 gene, subcloning the gene into plasmid pET30a, and its subsequent transfer to Escherichia coli with trigger factor 16 for co-expression. The protein thus expressed was purified with ammonium sulfate precipitation, hydrophobic chromatography, and endotoxin extraction procedures. VLPs were examined by transmission electron microscopy, dynamic light scattering, and the efficacy of VLPs vaccine was tested in vivo.

RESULTS

Results indicated that RDPV VP2 protein could be expressed soluble. Transmission electron microscopy and dynamic light scattering results indicated that RDPV VP2 self-assembled into VLPs. Hemagglutination inhibition antibody titers elicited by Al(OH) adjuvanted RDPV VLPs were comparable with RDPV inactivated vaccines, and the viral loads in the blood of the struck raccoon dogs were greatly reduced. Hematoxylin and eosin and Immunohistochemical results indicated that RDPV VLPs vaccine could protect raccoon dogs against RDPV infections.

CONCLUSIONS

These results suggest that RDPV VLPs can become a potential vaccine candidate for RDPV therapy.

摘要

背景

貉细小病毒(RDPV)可导致貉发生急性传染病,严重时可导致死亡。目前的治疗策略依赖于广泛使用经典的灭活疫苗,但存在剂量大、免疫周期短和安全性问题。

方法

本研究旨在优化 RDPV VP2 基因,将基因亚克隆到质粒 pET30a 中,并随后将其转移到带有触发因子 16 的大肠杆菌中进行共表达。表达的蛋白用硫酸铵沉淀、疏水层析和内毒素提取程序进行纯化。通过透射电子显微镜、动态光散射检查 VLPs,并在体内测试 VLPs 疫苗的功效。

结果

结果表明 RDPV VP2 蛋白可以表达为可溶性蛋白。透射电子显微镜和动态光散射结果表明 RDPV VP2 自我组装成 VLPs。用 Al(OH)佐剂的 RDPV VLPs 引起的血凝抑制抗体滴度与 RDPV 灭活疫苗相当,并且受感染的貉血液中的病毒载量大大降低。苏木精和伊红以及免疫组织化学结果表明,RDPV VLPs 疫苗可以保护貉免受 RDPV 感染。

结论

这些结果表明 RDPV VLPs 可以成为 RDPV 治疗的潜在疫苗候选物。

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