流感血凝素mRNA脂质纳米颗粒疫苗诱导呼吸道黏膜免疫
收稿日期: 2026-02-26
修回日期: 2026-03-26
录用日期: 2026-04-20
网络出版日期: 2026-06-29
基金资助
北京市自然科学基金资助项目（L258075）
An Influenza Hemagglutinin mRNA Lipid Nanoparticle Vaccine Elicits Respiratory Mucosal Immunity
Received date: 2026-02-26
Revised date: 2026-03-26
Accepted date: 2026-04-20
Online published: 2026-06-29
目的：研究mRNA黏膜流感疫苗，开发一种基于流感病毒血凝素（HA）的mRNA脂质纳米颗粒（LNPs），通过雾化吸入评估其免疫原性和安全性。方法：采用体外转录合成编码HA的mRNA，利用微流控技术制备包载HA mRNA的LNPs。以LNPs的理化性质和细胞转染效率为指标筛选最优处方，并考察其溶酶体逃逸能力、细胞摄取机制及细胞毒性。通过肌内注射与肺部雾化吸入方式免疫小鼠，比较其诱导的体液免疫与呼吸道黏膜免疫反应，并初步评价疫苗一般安全性。结果：合成的HA mRNA通过转染试剂转染细胞效率为77.6%，包载到LNPs中后，筛选得到的最优LNP处方（氮磷比8∶1）细胞转染效率达到90%以上。在细胞中，6 h内可实现有效的溶酶体逃逸，摄取主要依赖动力蛋白与小窝蛋白-1介导的内吞作用。雾化吸入免疫小鼠后，不仅能诱导高水平的HA特异性血清IgG抗体，还能诱导黏膜免疫IgA抗体的产生。免疫期间小鼠体重稳步增长，血清生化指标未见异常，重要脏器亦无明显病理损伤。结论：本研究合成的HA mRNA具有翻译表达蛋白能力，制备的最优HA mRNA-LNPs在体外具有良好的细胞递送效率与抗原表达能力，在体内可同时激发强效的体液和黏膜双重免疫，且安全性良好。
娄经虎
,
吴智能
,
程艺
,
李蒙
,
刘楠
,
王增明
,
高翔
,
郑爱萍
,
张慧
.
流感血凝素mRNA脂质纳米颗粒疫苗诱导呼吸道黏膜免疫
Objective： To investigate mRNA mucosal influenza vaccines， develop mRNA lipid nanoparticles （LNPs） based on influenza virus hemagglutinin （HA）， and assess their immunogenicity and safety through nebulized inhalation.Methods： mRNA encoding HA was synthesized via in vitro transcription， and LNPs encapsulating HA mRNA were prepared using microfluidic technology. The optimal formulation was identified based on the physicochemical properties and cell transfection efficiency of the LNPs. Additionally， the lysosomal escape capability， cell uptake mechanisms， and cytotoxicity of the LNPs were investigated. Mice were immunized via both intramuscular injection and pulmonary nebulized inhalation. The humoral and respiratory mucosal immune responses elicited were compared， and the general safety of the vaccine was preliminarily assessed.Results： The transfection efficiency of the synthesized HA mRNA in cells using transfection reagents was 77.6%. Following encapsulation in LNPs， the cell transfection efficiency of the optimal LNP formulation， characterized by a nitrogen-phosphorus ratio of 8∶1， exceeded 90%. Effective lysosomal escape occurred within 6 h， with the uptake pathway reliant on endocytosis mediated by dynamin and caveolin-1. After nebulized inhalation immunization in mice， high levels of HA-specific serum IgG antibodies were induced， alongside the production of mucosal immune IgA antibodies. Throughout the immunization period， the mice exhibited a steady increase in body weight， with no abnormalities detected in serum biochemical indicators and no significant pathological damage observed in vital organs.Conclusion： The HA mRNA synthesized in this study demonstrates the capacity to translate and express proteins. The optimal HA mRNA-LNPs that were prepared and screened exhibit excellent cell delivery efficiency and antigen expression capability in vitro， effectively stimulating robust humoral and mucosal dual immunity in vivo， while also ensuring good safety.
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