狂犬病病毒载体新冠口服或肌注式灭活疫苗
可保护水貂阻断新冠病毒的感染和传播

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

狂犬病病毒载体新冠口服或肌注式灭活疫苗可保护水貂阻断新冠病毒的感染和传播

收稿日期:2024-02-20 接受日期:2024-05-28 出版日期:2025-03-20 发布日期:2025-02-28

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

Received:2024-02-20 Accepted:2024-05-28 Online:2025-03-20 Published:2025-02-28
About author: #Correspondence Zhigao Bu, E-mail: buzhigao@caas.cn; Lei Shuai, E-mail: shuailei@caas.cn * These authors contributed equally to this work.
Supported by:
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).

摘要/Abstract

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.

Key words: SARS-CoV-2 , rabies virus , oral vaccine , inactivated vaccine , mink

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. 狂犬病病毒载体新冠口服或肌注式灭活疫苗可保护水貂阻断新冠病毒的感染和传播[J]. Journal of Integrative Agriculture, 2025, 24(3): 1198-1211.

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. Rabies virus-based oral and inactivated vaccines protect minks against SARS-CoV-2 infection and transmission[J]. Journal of Integrative Agriculture, 2025, 24(3): 1198-1211.

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狂犬病病毒载体新冠口服或肌注式灭活疫苗可保护水貂阻断新冠病毒的感染和传播

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

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狂犬病病毒载体新冠口服或肌注式灭活疫苗 可保护水貂阻断新冠病毒的感染和传播

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

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狂犬病病毒载体新冠口服或肌注式灭活疫苗可保护水貂阻断新冠病毒的感染和传播