Construction and evaluation of a universal influenza mRNA vaccine

Yuying TIAN; Zhuoya DENG; Cong LI; Fang SUN; Rui CAO; Penghui YANG

doi:10.16016/j.2097-0927.202312014

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Construction and evaluation of a universal influenza mRNA vaccine

Basic Medicine | 更新时间：2026-01-09

- Construction and evaluation of a universal influenza mRNA vaccine
- Journal of Army Medical University Vol. 46, Issue 7, Pages: 725-731(2024)
- 作者机构：
  
  1.010110 呼和浩特, 内蒙古医科大学基础医学院
  2.100853 北京, 中国人民解放军总医院第一医学中心肝胆胰外科医学部研究所
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202312014
  CLC： R392.2;R392.7;R511.7
- Received：04 December 2023，
  
  Revised：2024-01-16，
  
  Published：15 April 2024
- 稿件说明：
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Yuying TIAN, Zhuoya DENG, Cong LI, et al. Construction and evaluation of a universal influenza mRNA vaccine[J]. Journal of Army Medical University, 2024, 46(7): 725-731.
DOI：

Yuying TIAN, Zhuoya DENG, Cong LI, et al. Construction and evaluation of a universal influenza mRNA vaccine[J]. Journal of Army Medical University, 2024, 46(7): 725-731. DOI： 10.16016/j.2097-0927.202312014.

摘要

目的

构建通用流感mRNA疫苗并全面评价其免疫保护效果。

方法

优化流感病毒株A/California/04/2009的血凝素(hemagglutinin

HA)、核蛋白(nucleoprotein

NP)和基质蛋白2胞外区(matrix protein 2 ectodomain

M2e)抗原序列

并将HA、NP和3个串联的M2e (3M2e)分别克隆至pcDNA3.1载体

通

过线性化、体外转录、酶学法加帽、酶促加尾合成mRNA

命名为mRNA-HA、mRNA-NP和mRNA-3M2e。3种mRNA分别转染293T细胞后

通过免疫荧光实验鉴定蛋白的表达。利用脂质纳米颗粒分别包裹mRNA-HA、mRNA-NP和mRNA-3M2e并测量其粒径和电位。再将3种mRNA等体积混合制备成Comb-mRNA疫苗。将28只6周雌性Balb/c小鼠(体质量为18~22 g)按简单随机分组法分为2组: LNP组(

=14)和Comb-mRNA组(

=14)。通过血凝抑制(hemagglutination inhibition

HI)、微量中和(microneutralization

MN)实验评价流感mRNA疫苗诱导小鼠产生的血清抗体滴度; 通过流式细胞术评价Comb-mRNA疫苗诱导的细胞免疫反应。采用5LD

野生型H1N1流感病毒株感染小鼠评价Comb-mRNA疫苗的免疫保护效果。

结果

成功构建mRNA-HA、mRNA-NP和mRNA-3M2e

且3种mRNA均能在293T细胞表达。脂质纳米颗粒包裹mRNA平均粒径为(119.53±6.5) nm

平均电位为(-8.23±1.3) mV。与LNP组比较

Comb-mRNA组疫苗的HI几何平均滴度(geometric mean titer

GMT)达179.6

MN的GMT达201.6

同时诱导IFNγ

CD4

/CD8

T细胞比例升高。Comb-mRNA组在加强免疫后2周能够提供对5LD

野生流感H1N1亚型病毒的保护。

结论

通用流感疫苗候选疫苗Comb-mRNA能够诱导小鼠免疫反应并保护小鼠免受病毒感染。

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 (

=14) and Comb-mRNA group (

=14).Hemagglutination inhibition (HI) method and microneutralization (MN) test were used to evaluate the s

erum antibody titer induced by Comb-mRNA vaccines.The mice were infected by 5LD

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 5LD

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.

关键词

Keywords

references

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