摘 要：猴痘病毒（MPXV）的持续流行对全球公共卫生构成威胁，因此亟需开发高效、安全且具高度特异性的新型疫苗。本研究基于自组装纳米颗粒mi3平台，设计并构建了一种多价猴痘亚单位疫苗，旨在针对MPXV的4个关键抗原（A35R、B6R、E8L和M1R）进行协同免疫。通过密码子优化，在HEK-293F细胞中经重组表达并纯化后获得上述抗原蛋白，进一步利用SpyTag-SpyCatcher共价连接系统将其高效、定向展示于mi3纳米颗粒表面，形成结构稳定的纳米颗粒疫苗（NP组），同时设立蛋白混合组（SU组）作为对照。在BALB/c小鼠模型中，研究团队系统评估了疫苗诱导的体液与细胞免疫应答。结果表明，所制备的纳米颗粒疫苗粒径均一、结构完整。动物免疫试验显示，与SU组相比，NP组能诱导更高水平的抗原特异性IgG抗体（P<0.01），并激起强烈的Th1/Th2混合型免疫反应，体现为多种细胞因子（包括IL-2、IL-4、IL-5和IL-6）分泌水平显著升高。转录组分析进一步提示，该纳米颗粒疫苗可能通过调控IL-17等相关信号通路增强机体免疫应答。综上所述，本研究成功研发了一种基于mi3纳米颗粒的MPXV多价亚单位候选疫苗，并在小鼠模型中证明其具有良好的免疫原性，为下一代猴痘疫苗的研发提供了重要的试验依据与技术平台。
关键词：猴痘病毒；亚单位疫苗；纳米颗粒；mi3；免疫原性
中图分类号：R373.1         文献标志码：A     DOI：10.3969/j.issn.1007-7146.2026.03.004

Abstract: The ongoing global spread of monkeypox virus (MPXV) poses a persistent public health threat, underscoring the urgent need for novel vaccines that are effective, safe, and highly specific. In this study, we designed and constructed a multivalent subunit vaccine based on the self-assembling mi3 nanoparticle platform, targeting four key MPXV antigens (A35R, B6R, E8L, and M1R) for synergistic immunization. Following codon optimization, the antigen proteins were recombinantly expressed in HEK-293F cells and purified. Using the SpyTag-SpyCatcher covalent conjugation system, these antigens were efficiently and site-specifically displayed on the surface of mi3 nanoparticles, resulting in structurally stable nanovaccines (NP group). A soluble protein mixture group (SU group) was included as a control. Systemic humoral and cellular immune responses were evaluated in BALB/c mouse models. The prepared nanoparticle vaccines showed uniform size distribution and structural integrity. Immunization experiments revealed that the NP group induced significantly higher levels of antigen-specific IgG antibodies compared to the SU group (P<0.01) and elicited a potent Th1/Th2-mixed immune response, as reflected by markedly elevated secretion of multiple cytokines including IL-2, IL-4, IL-5, and IL-6. Transcriptomic analysis further suggested that the nanovaccine may enhance immune responses through modulation of key signaling pathways such as IL-17. In summary, we successfully developed a mi3 nanoparticle-based multivalent subunit vaccine candidate against MPXV, which demonstrated strong immunogenicity in a mouse model, providing an important experimental foundation and a technical platform for the development of next-generation monkeypox vaccines.
Keywords: monkeypox virus; subunit vaccine; nanoparticle; mi3; immunogenicity
（Acta Laser Biology Sinica, 2026, 35(3): 222-232）