Preparation and immunogenicity evaluation of a tick-borne encephalitis mRNA vaccine producing virus-like particle
10.13200/j.cnki.cjb.004535
- VernacularTitle:表达病毒样颗粒的森林脑炎mRNA疫苗的制备及其免疫原性评价
- Author:
LI Zhuang
- Publication Type:Journal Article
- Keywords:
Tick-borne encephalitis;
mRNA vaccine;
Virus-like particles(VLPs);
Immunogenicity
- From:
Chinese Journal of Biologicals
2025;38(08):897-901+907
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare an mRNA vaccine against tick-borne encephalitis virus(TBEV), verify its ability to produce virus-like particles(VLPs) in cells, and evaluate its immunogenicity in mice.Methods The gene sequence containing Japanese encephalitis virus(JEV) signal peptide, TBEV premature membrane glycoprotein(prM) and E protein was designed. The plasmids were constructed according to the sequence, and corresponding mRNA was synthesized by in-vitro transcription. The mRNA was transfected into HEK293T cells and then measured for the expression in cells by Western blot,and detected for the ability to self-assemble into VLPs in cells by electron microscopy. The mRNA lipid nanoparticles(LNPs)were prepared by Nano Assembler system, and the particle size and distribution of LNPs were measured. Female BALB/c mice were immunized by intramuscular injection at dosages of 2 and 10 μg mRNA vaccine respectively, 6 mice in each group, and the negative control group(PBS) was set. One week after booster immunization, orbital blood samples were collected and separated for serum. The binding antibody levels were detected by ELISA, and the neutralizing antibody levels were detected by plaque reduction assay.Results After mRNA transfection into HEK293T cells, a single band was found at the relative molecular mass of about 55 000, which was consistent with the theoretical relative molecular mass of E protein.Under electron microscope, spherical particles with a diameter of about 50 nm were observed, and there were obvious spikes on the surface. The average particle size of mRNA LNPs vaccine was 124. 4 nm and the polymer dispersity index(PDI) was0. 049. Compared with the negative control group, the levels of binding antibody and neutralizing antibody induced by 2 and10 μg mRNA significantly increased(t = 12. 4 and 10. 8, 19. 7 and 12. 0, respectively, each P < 0. 001).Conclusion A TBEV mRNA vaccine capable of producing VLPs in cells was successfully prepared, which demonstrated good immunogenicity in mice, providing experimental basis for the development of new generation TBEV vaccine in combination using of VLPs technology and mRNA technology.
