MRNA Vaccines Encoding the HA Protein of Influenza A H1N1 Virus Delivered by Cationic Lipid Nanoparticles Induce Protective Immune Responses in Mice.

Fulong Nan,Maopeng Wang,Xinyu Zhuang,Ningyi Jin,Yanxin Qi,He Zhang,Huijun Lu,Chang Li,Mingyao Tian,Ning Yu

doi:10.3390/vaccines8010123

Abstract

The design of the mRNA vaccine involves the selection of in vitro transcription (IVT) systems and nonviral delivery vectors. This study aimed to verify the effect of 5’ and 3’ untranslated region (UTR) sequences on the translation efficiency of mRNA. Three modes of IVT-mRNA systems (IVT-mRNA-n1/n2/n3) with diverse UTRs were constructed, and EGFP (enhanced green fluorescent protein) and HA (hemagglutinin) gene of H3N2 influenza virus were introduced into each of them. The results showed that the mode of 5’ and 3’ UTRs originating from human β-globulin was better than the mode of UTRs from human α-globulin, and the n3 mode was the best. mEGFP-n3, mH3HA-n3, and mLuciferease-n3 were prepared to compare the effect of cationic lipid nanoparticle (LNP) with that of mannose-conjugated LNP (LNP-Man) on the efficiency of gene delivery. The results showed that the effect of LNP-Man was better than that of LNP both in vitro and in vivo. Choosing appropriate ligands might help in vaccine design. After selecting the IVT-mRNA-n3 system and delivery vectors, mRNA vaccines were constructed against the H1N1 influenza virus, and C57BL/6 mice were immunized through intranasal administration. The results showed that mRNA vaccines could elicit both humoral and cellular immune responses and completely protect mice from the tenfold LD50 H1N1 influenza virus challenge.

Highlights

Influenza viruses belong to the family of Orthomyxoviridae, with negative-sence, single-stranded, and segmental RNA genomes
BHK-21, MDCK, HeLa, HEK-293, BEAS-2B, and NCI-H226 cells were transfected with mEGFP-n3 to verify the effect of the in vitro transcription (IVT)-mRNA-n3 system on other cell lines
5’ and 3’ untranslated region (UTR) originating from human β-globulin were better than those from human α-globulin in the design of the IVT-mRNA system

Summary

Introduction

Influenza viruses belong to the family of Orthomyxoviridae, with negative-sence, single-stranded, and segmental RNA genomes. Influenza A and B viruses cause threats to human health they can give rise to mild-to-severe respiratory infections. According to the statistics of the World Health Organization, seasonal influenza viruses (A/H1N1/H3N2), as well as influenza B viruses (B/Yamagata/Victoria), Vaccines 2020, 8, 123; doi:10.3390/vaccines8010123 www.mdpi.com/journal/vaccines. Vaccination is the best protection measure to prevent influenza. Seasonal influenza viruses can evolve constantly through antigen drift and antigen shift. These punctuated antigenic changes, especially in HA (the main target of protective antibodies), lead to evasion of the immunity elicited by previous infection or vaccination [2,3]. Vaccine strains need to be renewed annually [4]

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