Cui Junru, Yuan Fangfeng, Qin Jane, Jeon Ju Hyeong, Yun Dong Soo, Wang Tianlei, Xu Renhuan, Cao Hong, Tungate Ashleigh A, Netherton Christopher L, Chen Jianzhu
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.
Advanced RNA Vaccine Technologies, Inc., North Bethesda, Rockville, Maryland 20852, United States.
ACS Infect Dis. 2025 Aug 8;11(8):2104-2115. doi: 10.1021/acsinfecdis.5c00067. Epub 2025 Jul 9.
Viral capsid proteins are widely explored for subunit vaccine development but are often hampered by their complexity of production and low immunogenicity. Here, we report a simple approach to overcoming these challenges by combining mRNA vaccine technology with protein engineering. Using African swine fever virus (ASFV) capsid proteins P72 and penton as models, we engineered them into membrane-bound and secreted forms and compared their immunogenicity to that of the native intracellular form in mice and pigs through mRNA vaccination. The membrane-bound and secreted P72 and penton folded into their native multimeric structure independent of the viral chaperone, therefore preserving their conformational epitopes. The membrane-bound P72 and penton also elicited significantly stronger antibody and T cell responses than their secreted or intracellular counterparts. Our study provides a simple approach to enhancing folding, multimeric structure formation, and immunogenicity of viral capsid proteins for ASFV subunit vaccine development and immunogenicity of intracellular proteins in general.
病毒衣壳蛋白在亚单位疫苗开发中得到了广泛探索，但它们的生产复杂性和低免疫原性常常阻碍其发展。在此，我们报告一种通过将mRNA疫苗技术与蛋白质工程相结合来克服这些挑战的简单方法。以非洲猪瘟病毒（ASFV）衣壳蛋白P72和五邻体为模型，我们将它们设计成膜结合形式和分泌形式，并通过mRNA疫苗接种在小鼠和猪中比较它们与天然细胞内形式的免疫原性。膜结合型和分泌型P72和五邻体折叠成其天然多聚体结构，不依赖病毒伴侣蛋白，因此保留了它们的构象表位。膜结合型P72和五邻体也比其分泌型或细胞内型对应物引发了显著更强的抗体和T细胞反应。我们的研究为增强病毒衣壳蛋白的折叠、多聚体结构形成及免疫原性提供了一种简单方法，可用于ASFV亚单位疫苗开发及一般细胞内蛋白的免疫原性研究。
