Immune efficacy of 3M2e-based universal influenza DNA vaccine delivered by cell penetrating peptides in mice
10.13200/j.cnki.cjb.004291
- VernacularTitle:细胞穿透肽递送的流感病毒M2胞外区通用DNA疫苗在小鼠体内的免疫效力
- Author:
LIU Honggang
- Publication Type:Journal Article
- Keywords:
Influenza A virus;
M2 extracellular domain;
Universal DNA vaccine;
Cell penetrating peptides(CPPs);
Cross protection
- From:
Chinese Journal of Biologicals
2024;37(9):1030-1036
- CountryChina
- Language:Chinese
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Abstract:
Objective To design an effective universal DNA vaccine for influenza A virus and detect its immune efficacy in mice.Methods The DNA sequence containing the M2(3M2e)extracellular domain of H1N1,H3N2 and H9N2 subtypes of influenza A virus was synthesized and cloned into vector pVAX1 to construct eukaryotic expression plasmid pVAX1-3M2e. The plasmid pVAX1-3M2e was mixed with cell penetrating peptides(CPPs)RVG9dR,Protamine,and RVG9dR +Protamine peptide complex(1∶1)at different mass fractions(1∶0. 5,1∶1,1∶2,1∶4,and 1∶8),separately,and gel retardation was performed to determine the proper binding of DNA and CPPs. The delivery efficiency of the CPPs was detected by cell fluorescence assay at cellular level(293T cells). Male BALB/c mice were immunized intramuscularly with plasmid pVAX1-3M2e delivered by RVG9dR + Protamine(two doses:25 μg pVAX1-3M2e + 500 μg polypeptide complex,50 μg pVAX1-3M2e + 1 000 μg polypeptide complex,8 mice in each group)to detect the cross-protection effect of universal influenza A virus DNA vaccine against three subtypes of influenza viruses(H1N1,H3N2 and H9N2).Results The optimum binding ratios of DNA to RVG9dR,Protamine,and RVG9dR + Protamine were 1∶2,1∶2,and 1∶1,respectively. RVG9dR + Protamine had superior delivery efficiency to plasmid pVAX1-3M2e at the mass fraction of 1∶20. The protective rates of 50 μg pVAX1-3M2e + 1 000 μg polypeptide complex against three subtypes of influenza virus in mice were all 100%.Conclusion The plasmid pVAX1-3M2e delivered by RVG9dR + Protamine polypeptide complex provides cross protection against three subtypes of influenza A virus in mice.
