Rabies virus-based oral and inactivated vaccines protect minks against SARS-CoV-2 infection and transmission

doi:10.1016/j.jia.2024.07.024

Journal of Integrative Agriculture

2025, Vol. 24

Issue (3): 1198-1211 DOI: 10.1016/j.jia.2024.07.024

Animal Science · Veterinary Medicine

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Rabies virus-based oral and inactivated vaccines protect minks against SARS-CoV-2 infection and transmission

Hong Huo^{1, 2*}, Shuang Xiao^1*, Jinming Wang^1*, Xijun Wang¹, Jinying Ge¹, Gongxun Zhong^{1, 2}, Zhiyuan Wen^{1, 2}, Chong Wang^{1, 2}, Jinliang Wang^{1, 2}, Han Wang¹, Xijun He², Lei Shuai^{1, 2#}, Zhigao Bu^{1, 2, 3#}

¹State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China

²National High Containment Laboratory for Animal Diseases Control and Prevention, Harbin 150069, China

³Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China

Highlights
● Recombinant rabies virus expressing the prefusion-stabilized SARS-CoV-2 spike protein as an oral or inactivated vaccine induced strong humoral immune responses against SARS-CoV-2 strains in mice and minks.
● rERAG_333E-S6P completely protected mice and minks against SARS-CoV-2 infection.
● rERAG_333E-S6P can largely protect minks against wild-type SARS-CoV-2 transmission via respiratory droplets.

Abstract
References

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摘要

新型冠状病毒感染（COVID-19）是一种由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的人兽共患病，对人类健康和公共卫生安全造成极大的影响。SARS-CoV-2易感动物范围广泛，其中水貂对SARS-CoV-2高度敏感，并可将SARS-CoV-2传播给人类。动物COVID-19疫苗的研发和应用是降低和阻断动物SARS-CoV-2感染和传播的有效手段。对于水貂和野生动物的新冠感染防控，口服免疫是预防SARS-CoV-2感染和传播的有效策略之一。

本研究以SARS-CoV-2原始株（WT）、德尔塔株（Delta）和奥密克戎BA.2（BA.2）株刺突蛋白（S）基因为靶基因，分别进行预融合、结构稳定等方面分子修饰，并人工合成可表达S6P、DS6P、BA2S6P蛋白的S基因。将合成的基因插入狂犬病病毒株（rabies virus, RABV）rERAG_333E的P和M基因之间，构建重组RABV病毒rERAG_333E/S6P、rERAG_333E/DS6P、rERAG_333E/BA2S6P。重组病毒体外感染试验结果显示，S6P、DS6P、BA2S6P均可稳定、高效表达预融合的S6P、DS6P、BA2S6P蛋白，并且不影响重组病毒的体外生长性状。重组RABVs作为口服疫苗分别单独或联合免疫BALB/c小鼠，体重变化与载体疫苗免疫组相比无显著差异，并能诱导产生高水平的SARS-CoV-2中和抗体、WT/Delta/BA.2交叉中和抗体反应、唾液sIgA反应及RABV中和抗体反应；作为肌注式灭活疫苗单独或联合免疫BALB/c小鼠，同样显示出优秀的安全性和免疫原性。此外，rERAG_333E/S6P作为口服、肌注式灭活疫苗均可有效阻止SARS-CoV-2在BALB/c小鼠上、下呼吸道的复制。

为评估重组RABVs作为新冠易感动物新冠疫苗的潜力，本研究将重组RABVs作为口服疫苗或肌注式灭活疫苗单独或联合免疫水貂，监测期内水貂均健活，且具有良好的免疫原性，联合免疫可诱导高效的SARS-CoV-2 WT/Delta/Omicron BA.2株交叉中和抗体反应及RABV中和抗体反应。水貂攻毒保护和传播阻断试验结果显示，rERAG_333E/S6P作为口服、肌注式灭活疫苗在水貂上均具有良好的SARS-CoV-2 WT株攻毒保护和传播阻断效果，可有效降低SARS-CoV-2对水貂的感染、肺脏损伤和传播。

本研究结果表明，表达SARS-CoV-2修饰S蛋白的重组RABVs可作为口服疫苗候选株应用于水貂、流浪动物、野生动物等，作为肌注式灭活疫苗候选株应用于伴侣动物；联合免疫的免疫策略有潜力成为抵抗多种SARS-CoV-2流行株的动物用COVID-19多价疫苗；借助RABV灭活疫苗的成熟生产工艺，有潜力作为人用COVID-19和狂犬病二联候选疫苗株。

Abstract

Minks are highly susceptible to SARS-CoV-2, and have transmitted SARS-CoV-2 to humans. Oral immunization is one of the most promising strategies to prevent SARS-CoV-2 infection and transmission in minks. Here, we generated 3 recombinant rabies viruses (RABV), rERAG_333E/S6P, rERAG_333E/DS6P and rERAG_333E/BA2S6P, expressing the prefusion-stabilized SARS-CoV-2 spike protein of wild-type (S6P), δ (DS6P) or BA.2 (BA2S6P) strain based on an oral rabies vaccine candidate (rERAG_333E). Oral or inactivated immunization of the 3 RABVs monovalent or trivalent were safe, and induced robust RABV neutralizing antibody and cross-antibody responses against the three SARS-CoV-2 in mice and minks. The challenge tests showed that 2 doses of rERAG_333E-S6P as an oral or inactivated vaccine completely protected mice against mouse-adapted SARS-CoV-2 infection in the upper and lower respiratory tracts, and largely prevented viral replication and lung damage caused by wild-type SARS-CoV-2 infection in minks. Notably, we also confirmed that 2 doses of rERAG_333E-S6P as an oral or inactivated vaccine can largely protect minks against wild-type SARS-CoV-2 transmission via respiratory droplets. Our findings suggest that rERAG_333E-based COVID-19 vaccines appear to be suitable oral candidates to protect minks from SARS-CoV-2 infection and transmission, and may serve as inactivated vaccines for further investigation in humans.

Keywords: SARS-CoV-2 rabies virus oral vaccine inactivated vaccine mink

Received: 20 February 2024 Accepted: 28 May 2024

Fund:

This study was supported by the National Key Research and Development Program of China (2021YFC2301700), and the Natural Science Foundation of Heilongjiang Province, China (YQ2022C040).

About author: #Correspondence Zhigao Bu, E-mail: buzhigao@caas.cn; Lei Shuai, E-mail: shuailei@caas.cn * These authors contributed equally to this work.

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Hong Huo, Shuang Xiao, Jinming Wang, Xijun Wang, Jinying Ge, Gongxun Zhong, Zhiyuan Wen, Chong Wang, Jinliang Wang, Han Wang, Xijun He, Lei Shuai, Zhigao Bu. 2025. Rabies virus-based oral and inactivated vaccines protect minks against SARS-CoV-2 infection and transmission. Journal of Integrative Agriculture, 24(3): 1198-1211.

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