AbstractThe development of broad-spectrum coronavirus vaccines is essential to prepare for future respiratory virus pandemics. We demonstrated broad neutralization by a trivalent subunit vaccine, formulating the receptor-binding domains of SARS-CoV, MERS-CoV, and SARS-CoV-2 XBB.1.5 with Alum and CpG55.2.

摘要广谱冠状病毒疫苗的开发对于准备未来的呼吸道病毒大流行至关重要。我们证明了三价亚单位疫苗的广泛中和作用，用明矾和CpG55.2配制了SARS-CoV，MERS-CoV和SARS-CoV-2 XBB.1.5的受体结合结构域。

Vaccinated mice produced cross-neutralizing antibodies against all three human Betacoronaviruses and others currently exclusive to bats, indicating the epitope preservation of the individual antigens during co-formulation and the potential for epitope broadening..

接种疫苗的小鼠产生了针对所有三种人类Betacoronaviruses和目前蝙蝠独有的其他病毒的交叉中和抗体，表明在共同配制过程中单个抗原的表位保存以及表位扩大的潜力。。

Given the emergence of three highly pathogenic and deadly human Betacoronaviruses—severe acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome (MERS)-CoV, and SARS-CoV-2— within the past two decades, together with the looming threat of additional zoonotic viral spillovers, creating a broadly protective coronavirus vaccine capable of safeguarding against the five major lineages of the virus (Embecovirus, Sarbecovirus, Merbecovirus, Nobecovirus, and Hibecovirus)1 remains an urgent global health challenge.

鉴于在过去二十年中出现了三种高致病性和致命性的人类β-冠状病毒-严重急性呼吸综合征（SARS）-冠状病毒，中东呼吸综合征（MERS）-冠状病毒和SARS-CoV-2-以及其他人畜共患病毒溢出的威胁，创造了一种广泛保护的冠状病毒疫苗，能够抵抗该病毒的五个主要谱系（Embecovirus，Sarbecovirus，Merbecovirus，Nobecovirus和Hibecovirus）1仍然是一个紧迫的全球健康挑战。

Compared to current vaccines, which are either species- or variant-specific, a more broadly protective pan-coronavirus vaccine could provide considerable epidemiological, clinical, and economic value to mitigate the global disease burden2.Early during the COVID-19 pandemic, the development of broadly protective coronavirus vaccines was motivated by the observation of cross-clade Sarbecovirus neutralizing antibodies in previously SARS-CoV-infected and then Pfizer BioNTech BNT162b2-vaccinated individuals3.

与目前的物种或变异特异性疫苗相比，更广泛的保护性泛冠状病毒疫苗可以提供相当大的流行病学，临床和经济价值，以减轻全球疾病负担2。在COVID-19大流行早期，广泛保护性冠状病毒疫苗的开发是由于在先前感染SARS-CoV的个体中观察到交叉进化枝沙伯病毒中和抗体，然后是辉瑞生物技术公司BNT162b2接种的个体3。

Now, with support from national and international funding agencies, multiple broadly protective vaccines are under development4,5.We focused our coronavirus vaccine development efforts on low-cost, easy-to-produce RBD-based vaccines6. In 2013, we developed RBD219-N1 as a SARS-CoV vaccine that protected in a mouse challenge model7, and when MERS-CoV emerged, we produced a MERS RBD vaccine that protected mice against a lethal MERS-CoV infection8,9.

现在，在国家和国际资助机构的支持下，正在开发多种广泛保护性疫苗4,5。我们将冠状病毒疫苗的开发工作重点放在低成本，易于生产的RBD疫苗6上。2013年，我们开发了RBD219-N1作为SARS-CoV疫苗，可在小鼠攻击模型中保护7，当MERS-CoV出现时，我们生产了一种MERS-RBD疫苗，可保护小鼠免受致命的MERS-CoV感染8,9。

In 2020, our RBD-based SARS-CoV-2 vaccine was efficacious in various animal models10,11 and transitioned into the clinic, where it was proven safe, immunogenic, and effective12. With our vaccine technology, two vaccines (Corbevax, Biological E Ltd, India; and Indovac, Bio Farma, Indonesia) were produced for les.

2020年，我们基于RBD的SARS-CoV-2疫苗在各种动物模型中都是有效的10,11，并过渡到临床，在那里它被证明是安全的，免疫原性的和有效的12。利用我们的疫苗技术，为les生产了两种疫苗（Corbevax，Biological E Ltd，印度；Indovac，Bio Farma，印度尼西亚）。

Data availability

数据可用性

The data supporting this study’s findings are available from the corresponding author upon reasonable request.

根据合理的要求，通讯作者可以提供支持本研究结果的数据。

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Download referencesAcknowledgementsThis work was supported by Texas Children’s Hospital Center for Vaccine Development Intramural Funds, USA. In addition, we gratefully acknowledge the donations from various private individuals received in support of our vaccine development program. We thank Dr.

下载参考文献致谢这项工作得到了美国德克萨斯州儿童医院疫苗开发内部基金会的支持。此外，我们非常感谢为支持我们的疫苗开发计划而收到的各种私人捐款。我们感谢博士。

Nikolai Petrovsky (Vaxine Pty Ltd, Australia) for kindly sharing the CpG55.2 adjuvant. We thank Vincent Munster (NIAID) for providing the spike expression plasmid for SARS-CoV-2. All the authors would also like to extend their sincere gratitude to Diane Niño, Allan Ciciriello, and Kay Razavi for their invaluable contributions to the vaccine center’s COVID-19 vaccine program.Author informationAuthors and AffiliationsTexas Children’s Hospital Center for Vaccine Development, Houston, TX, 77030, USASyamala Rani Thimmiraju, Rakesh Adhikari, JeAnna R.

尼古拉·彼得罗夫斯基（澳大利亚Vaxine私人有限公司），感谢他分享CpG55.2佐剂。我们感谢Vincent Munster（NIAID）为SARS-CoV-2提供了尖峰表达质粒。所有作者还要衷心感谢黛安·尼诺，艾伦·西西里略和凯·拉扎维为疫苗中心的新型冠状病毒肺炎疫苗计划做出的宝贵贡献。作者信息作者和附属机构德克萨斯州休斯顿特克斯儿童医院疫苗开发中心，77030，USASyamala Rani Thimmiraju，Rakesh Adhikari，JeAnna R。

Redd, Maria Jose Villar, Jungsoon Lee, Zhuyun Liu, Yi-Lin Chen, Amandeep Kaur, Nestor L. Uzcategui, Shannon E. Ronca, Wen-Hsiang Chen, Bin Zhan, Ulrich Strych, Maria Elena Bottazzi, Peter J. Hotez & Jeroen PolletDepartment of Pediatrics, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, 77030, USASyamala Rani Thimmiraju, Rakesh Adhikari, JeAnna R.

里德（Redd）、玛利亚·何塞·维拉尔（MariaJoseVillar）、李俊顺（Jungsoon Lee）、刘朱云（Zhuyun Liu）、陈宜林（Yi Lin Chen）、阿曼迪普·考尔（Amandeep Kaur）、内斯特·乌兹卡特吉（Nestor L.Uzcategui）、香农·E·朗卡（Shannon E.Ronca）、陈文祥（Wen Xiang Chen）、詹斌（Bin Zhan）、乌尔里希·斯特里奇（UlrichStrych）、玛利亚·埃琳娜·博塔齐（MariaElenaBottazzi）、彼得·霍特斯（PeterJ。

Redd, Maria Jose Villar, Jungsoon Lee, Zhuyun Liu, Yi-Lin Chen, Amandeep Kaur, Nestor L. Uzcategui, Shannon E. Ronca, Wen-Hsiang Chen, Bin Zhan, Ulrich Strych, Maria Elena Bottazzi, Peter J. Hotez & Jeroen PolletDepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, 77030, USASuman Sharma, Shannon E.

里德（Redd），玛利亚·何塞·维拉尔（MariaJoseVillar），李俊顺（Jungsoon Lee），刘朱云（Zhuyun Liu），陈宜林（Yi Lin Chen），阿曼迪普·考尔（Amandeep Kaur），内斯特·乌兹卡特吉（Nestor L.Uzcategui），香农·E·朗卡（Shannon E.Ronca），陈文祥（Wen Xiang Chen），詹斌（Bin Zhan），乌尔里希·斯特里奇（UlrichStrych），玛利亚·埃琳娜·博塔齐（Maria。

Ronca, Jason T. Kimata & Jeroen PolletDepartment of Biology, Baylor University, Waco, TX, 76706, USAMaria Elena Bottazzi & Peter J. HotezJames A. Baker III Institute for Public Policy, Rice University, Houston, TX, 77.

Ronca，Jason T.Kimata和Jeroen PolletDepartment of Biology，Baylor University，Waco，TX，76706，USAMaria Elena Bottazzi和Peter J.HotezJames A.Baker III公共政策研究所，莱斯大学，休斯顿，TX，77。

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PubMed Google ScholarContributionsConceptualization (Hypothesis and study design): S.R.T., W.-H.C., U.S., M.E.B., P.J.H., and J.P. Funding acquisition: M.E.B. and P.J.H. Lab experiments (Cloning): N.L.U., B.Z., S.S., J.T.K.; (Protein production): J.L., Z.L., A.K.; (Vaccine formulation): Y.-L.C.

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and J.P.; (Animal work): R.A., J.R.R., and S.E.R.; (Immunology): S.R.T., R.A., and M.J.V. Formal analysis (Data analysis): S.R.T., R.A., W.-H.C., J.T.K., B.Z., U.S., and J.P. Supervision (Final responsibility of teamwork) J.K., S.E.R., U.S., M.E.B., P.J.H., and J.P. Writing: S.R.T., J.T.K., U.S., and J.P.

和J.P。；（动物工作）：R.A.，J.R.R。和S.E.R。；（免疫学）：S.R.T.，R.A。和M.J.V.正式分析（数据分析）：S.R.T.，R.A.，W.H.C.，J.T.K.，B.Z.，美国和J.P.监督（团队合作的最终责任）J.K.，S.E.R.，U.S.，M.E.B.，P.J.H。和J.P.写作：S.R.T.，J.T.K.，U.S.，和J.P。

Visualization (Figures): S.R.T. and J.P. All authors have read, reviewed and agreed to the published version of the manuscript.Corresponding authorCorrespondence to.

可视化（数字）：S.R.T.和J.P.所有作者都阅读，审查并同意稿件的发布版本。对应作者对应。

Jeroen Pollet.Ethics declarations

杰伦·波莱特。道德宣言

Competing interests

相互竞争的利益

Several of the authors are co-inventors of a COVID-19 recombinant protein vaccine technology held by Baylor College of Medicine (BCM). BCM has licensed this technology non-exclusively to multiple companies dedicated to improving vaccine access in low- and middle-income countries, with no patent restrictions.

几位作者是贝勒医学院（BCM）持有的新型冠状病毒-19重组蛋白疫苗技术的共同发明人。BCM已将这项技术非独家许可给多家致力于改善中低收入国家疫苗获取的公司，没有专利限制。

The authors themselves are not directly involved in the licensing negotiations led by BCM. None of the authors have additional competing financial or non-financial interests..

作者本人并不直接参与BCM领导的许可谈判。没有一位作者有其他相互竞争的财务或非财务利益。。

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To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/..

要查看此许可证的副本，请访问http://creativecommons.org/licenses/by/4.0/..

Reprints and permissionsAbout this articleCite this articleThimmiraju, S.R., Adhikari, R., Redd, J.R. et al. A trivalent protein-based pan-Betacoronavirus vaccine elicits cross-neutralizing antibodies against a panel of coronavirus pseudoviruses.

转载和许可本文引用本文Thimmiraju，S.R.，Adhikari，R.，Redd，J.R。等人。一种基于三价蛋白质的泛β-冠状病毒疫苗引发针对一组冠状病毒假病毒的交叉中和抗体。

npj Vaccines 9, 132 (2024). https://doi.org/10.1038/s41541-024-00924-xDownload citationReceived: 28 February 2024Accepted: 12 July 2024Published: 22 July 2024DOI: https://doi.org/10.1038/s41541-024-00924-xShare this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

npj疫苗9132（2024）。https://doi.org/10.1038/s41541-024-00924-xDownload引文接收日期：2024年2月28日接受日期：2024年7月12日发布日期：2024年7月22日OI：https://doi.org/10.1038/s41541-024-00924-xShare本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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