@article{TIAN2024, 
author = {Yuying TIAN and Zhuoya DENG and Cong LI and Fang SUN and Rui CAO and Penghui YANG},
title = {Construction and evaluation of a universal influenza mRNA vaccine},
year = {2024},
journal = {Journal of Army Medical University},
volume = {46},
number = {7},
pages = {725-731},
keywords = {mRNA vaccine, influenza vaccine, hemagglutinin, nucleoprotein, matrix protein 2 ectodomain},
url = {https://www.sciopen.com/article/10.16016/j.2097-0927.202312014},
doi = {10.16016/j.2097-0927.202312014},
abstract = {ObjectiveTo construct a universal influenza mRNA vaccine and evaluate its immunogenicity.MethodsThe antigen sequence of hemagglutinin (HA), nucleoprotein (NP) and matrix protein 2 ectodomain (M2e) in influenza A/California/04/2009 was optimized. HA, NP and 3 tandem M2e (3M2e) were cloned into pcDNA3.1 vector, respectively. Then the mRNAs were synthesized by linearization, in vitro transcription, enzymatic capping and enzymatic tailing, and named as mRNA-HA, mRNA-NP and mRNA-3M2e, respectively. The protein expression of the 3 kinds of mRNAs in 293T cells was detected by immunofluorescence assay. Comb-mRNA vaccine was prepared by enveloped mRNA-HA, mRNA-NP and mRNA-3M2e with lipid nanoparticles, respectively, and the particle size and potential were identified. Twenty-eight 6-week-old female BALB/c mice (18~22 g) were randomly divided into LNP group (n=14) and Comb-mRNA group (n=14). Hemagglutination inhibition (HI) method and microneutralization (MN) test were used to evaluate the serum antibody titer induced by Comb-mRNA vaccines. The mice were infected by 5LD50 wild-type H1N1 influenza virus to evaluate the protective efficacy.ResultsThe mRNA-HA, mRNA-NP and mRNA-3M2e were successfully constructed, and the 3 mRNAs could be expressed in 293T cells. The average size of mRNA encapsulated by lipid nanoparticles was 119.53±6.5 nm, and the average potential was -8.23±1.3 mV. The geometric mean titer (GMT) of HI and MN in the Comb-mRNA group were 179.6 and 201.6, compared with the LNP group. The ratio of IFN-γ+CD4+/CD8+T cells was increased. The Comb-mRNA group could provide protection against 5LD50 wild type influenza H1N1 virus after 2 weeks of booster immunization.ConclusionComb-mRNA, an influenza vaccine candidate, can induce immune responses and protect mice from influenza virus challenge.}
}