Construction and evaluation of Bandavirus davieense virus mRNA vaccine
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1.Huadong Medical Institute of Biotechniques;2.LiuXinjian

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The Project of National Vaccine Innovation Platform

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    Abstract:

    Objective: Construction of Bandavirus davieense virus (DBV) vaccine based on the strategy of mRNA vaccine and evaluation the immunologic characteristics by immunizing BALB/c mice. Method: The coding sequence of DBV glycoprotein Gn was optimized and synthesized, then inserted into pGEM-3Zf(+) plasmid. The linearization plasmids were enzyme digested by BamH I, cap and polyA tailing were added with polymerase to complete in vitro transcription. Western blot verified the protein expression by transient transfection of the mRNA into 293FT cells. The mRNA was delivered by lipid nanoparticles, and immunized BALB/c mice in a low (2μg), middle (5μg) and high ( 20μg) dose once every two weeks,i.m. The antibody titer in mice serum was detected by ELISA, and the ability of antibody to neutralize DBV in vitro was detected at the cellular level by virus neutralization assay. Results: The mRNA obtained in the present study could transcript and express in vitro. It is also induced the mice to express high titer neutralizing antibodies in the low, middle and high dosage groups. Furthermore, the high dose group could induce the expression of the specific neutralizing antibodies more than 10 weeks. The virus neutralization assay showed that the antibodies in mice serum had the ability to avoid the virus infecting the cells. Conclusion: The DBV mRNA vaccine was successfully constructed, which may lay the basis for the further of preventing DBV infection.

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History
  • Received:December 21,2023
  • Revised:March 26,2024
  • Adopted:August 01,2024
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