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Publishing Language: Chinese

Construction and evaluation of a universal influenza mRNA vaccine

Yuying TIAN1,2, Zhuoya DENG2, Cong LI1,2, Fang SUN2, Rui CAO1,2, Penghui YANG1,2( )
College of Basic Medical Science, Inner Mongolia Medical University, Hohhot, Inner Mongolia Autonomous Region, 010110, China
Institute of Hepato-Pancreato-Biliary Surgery, The First Medical Center, Chinese PLA General Hospital, Beijing, 100853, China
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Abstract

Objective

To construct a universal influenza mRNA vaccine and evaluate its immunogenicity.

Methods

The 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.

Results

The 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.

Conclusion

Comb-mRNA, an influenza vaccine candidate, can induce immune responses and protect mice from influenza virus challenge.

Keywords

mRNA vaccineinfluenza vaccinehemagglutininnucleoproteinmatrix protein 2 ectodomain
CLC number: R392.2;R392.7;R511.7 Document code: A

References

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Journal of Army Medical University
Volume 46 Issue 7,
April 2024
Pages 725-731
DOI: 10.16016/j.2097-0927.202312014

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Cite this article:
TIAN Y, DENG Z, LI C, et al. Construction and evaluation of a universal influenza mRNA vaccine. Journal of Army Medical University, 2024, 46(7): 725-731. https://doi.org/10.16016/j.2097-0927.202312014

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Received: 04 December 2023
Revised: 16 January 2024
Published: 15 April 2024
© 2024 Journal of Army Medical University