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OBJECTIVE To investigate the immunogenicity and immune-protective efficiency of influenza A mRNA vaccines based on mRNA vaccine technology platform and provide new research strategies for the development of multivalent seasonal influenza mRNA vaccines. METHODS mRNA-H1N1 encapsulated by lipid nanoparticles(LNP) was transfected into HEK 293T cells, and the expression of LNP/mRNA-H1N1 was analyzed by Western blot and immunofluorescence microscopy in vitro, respectively. Furthermore, LNP/mRNA H1N1 was administered via intramuscular injection to SPF grade BALB/c mice, and the HA specific serum IgG titers induced by LNP/mRNA H1N1 were detected by ELISA. The secretion level of cytokine IFN-γ induced by LNP/mRNA-H1N1 was detected by ELISpot, HI titers induced by LNP/mRNA-H1N1 was measured by hemagglutination inhibition (HI) assay. Finally, the immune-protective efficacy LNP/mRNA-H1N1 was detected by the homologous H1N1 virus challenge experiment. RESULTS The specific expression of antigen protein HA could be detected by Western blot and fluorescence microscope, its molecular weight was approximately 75 000. Furthermore, LNP/mRNA-NH1N1 could induce significant humoral immune response, cellular immune response, and HI titers in BALB/c mice after prime and boost immunization. More importantly, LNP/mRNA-NH1N1 could provide 100% immune protection efficiency against H1N1 virus challenge. CONCLUSION LNP/mRNA-NH1N1 is an effective influenza mRNA candidate vaccine, laying the foundation for future preclinical safety evaluation trials in large animals and providing research ideas for the development of mRNA vaccines for other subtypes of influenza or other infectious disease vaccines.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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