人畜共患冠状病毒对人类健康构成持续威胁,新发现的蝙蝠传播病毒如猪急性腹泻综合征冠状病毒(SADS-CoV)导致仔猪高死亡率。体外研究表明,SADS-CoV可以感染来自不同物种的细胞系,包括人类,强调其对人类健康的潜在风险。然而,缺乏研究病毒进入的工具,随着疫苗或抗病毒治疗的缺乏,延续了这种威胁。为了解决这个问题,我们设计了水泡性口炎病毒(VSV)的感染性分子克隆,用SADS-CoV尖峰(S)替换其天然糖蛋白(G),并在3\'前导区插入Venus报告基因,以产生具有复制能力的rVSV-Venus-SADS病毒。rVSV-Venus-SADSS的连续传代导致在S蛋白的细胞质尾部中鉴定出11个氨基酸的截短,由于S蛋白的细胞膜锚定增加,这允许更有效的病毒繁殖。S蛋白整合到rVSV-Venus-SADSSΔ11颗粒中,对SADS-CoVS1蛋白免疫兔的血清中和敏感。此外,我们发现TMPRSS2促进SADS-CoV刺突介导的细胞进入。此外,我们使用初免-加强免疫策略评估了接种rVSV-Venus-SADSSΔ11的小鼠的血清中和能力,揭示针对SADS-CoV感染的有效中和抗体。总之,我们开发了一种安全实用的工具,用于研究SADS-CoV入室情况,并探索重组VSV载体SADS-CoV疫苗的潜力.重要的人畜共患冠状病毒,如猪急性腹泻综合征冠状病毒(SADS-CoV),对人类和动物健康构成持续威胁。为了解决这个问题,我们通过修改水泡性口炎病毒(VSV)设计了一个安全有效的工具,创造了一种具有复制能力的rVSV-金星-SADS病毒。通过连续通道,我们优化了病毒以增强膜锚定,病毒繁殖的关键因素。这种改良的病毒,rVSV-Venus-SADSSΔ11,被证明容易中和,为潜在的疫苗开辟道路。此外,我们的研究揭示了TMPRSS2在SADS-CoV进入中的作用。用rVSV-Venus-SADSSΔ11接种的小鼠产生了针对SADS-CoV的有效中和抗体。总之,我们的工作为研究SADS-CoV入门提供了一种安全实用的工具,并探索了重组VSV载体SADS-CoV疫苗的前景.
Zoonotic coronaviruses pose a continuous threat to human health, with newly identified bat-borne viruses like swine acute diarrhea syndrome coronavirus (SADS-CoV) causing high mortality in piglets. In vitro studies indicate that SADS-CoV can infect cell lines from diverse species, including humans, highlighting its potential risk to human health. However, the lack of tools to study viral entry, along with the absence of vaccines or antiviral therapies, perpetuates this threat. To address this, we engineered an infectious molecular clone of Vesicular Stomatitis Virus (VSV), replacing its native glycoprotein (G) with SADS-CoV spike (S) and inserting a Venus reporter at the 3\' leader region to generate a replication-competent rVSV-Venus-SADS S virus. Serial passages of rVSV-Venus-SADS S led to the identification of an 11-amino-acid truncation in the cytoplasmic tail of the S protein, which allowed more efficient viral propagation due to increased cell membrane anchoring of the S protein. The S protein was integrated into rVSV-Venus-SADS SΔ11 particles, susceptible to neutralization by sera from SADS-CoV S1 protein-immunized rabbits. Additionally, we found that TMPRSS2 promotes SADS-CoV spike-mediated cell entry. Furthermore, we assessed the serum-neutralizing ability of mice vaccinated with rVSV-Venus-SADS SΔ11 using a prime-boost immunization strategy, revealing effective neutralizing antibodies against SADS-CoV infection. In conclusion, we have developed a safe and practical tool for studying SADS-CoV entry and exploring the potential of a recombinant VSV-vectored SADS-CoV vaccine.IMPORTANCEZoonotic coronaviruses, like swine acute diarrhea syndrome coronavirus (SADS-CoV), pose a continual threat to human and animal health. To combat this, we engineered a safe and efficient tool by modifying the Vesicular Stomatitis Virus (VSV), creating a replication-competent rVSV-Venus-SADS S virus. Through serial passages, we optimized the virus for enhanced membrane anchoring, a key factor in viral propagation. This modified virus, rVSV-Venus-SADS SΔ11, proved susceptible to neutralization, opening avenues for potential vaccines. Additionally, our study revealed the role of TMPRSS2 in SADS-CoV entry. Mice vaccinated with rVSV-Venus-SADS SΔ11 developed potent neutralizing antibodies against SADS-CoV. In conclusion, our work presents a secure and practical tool for studying SADS-CoV entry and explores the promise of a recombinant VSV-vectored SADS-CoV vaccine.