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Novel coronavirus vaccine based on vaccinia virus TianTan strain

A vaccinia virus, coronavirus technology, applied in the direction of microorganism-based methods, viruses, viral peptides, etc., can solve problems such as the inability to eliminate the threat of the new coronavirus, and achieve the effect of solving the threat

Inactive Publication Date: 2021-09-03
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are a few antiviral drugs that have a certain effect on the infection of the new coronavirus, they cannot eliminate the threat of the new coronavirus to humans, so vaccination is still an effective measure for the new coronavirus

Method used

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  • Novel coronavirus vaccine based on vaccinia virus TianTan strain
  • Novel coronavirus vaccine based on vaccinia virus TianTan strain
  • Novel coronavirus vaccine based on vaccinia virus TianTan strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Construction of recombinant vaccinia virus TK-S capable of expressing the new crown S protein

[0023] 1. Method

[0024] 1.1. Construction of recombinant plasmid pTK-S

[0025] After codon-optimizing the spike protein S coding sequence of the new coronavirus SARS-CoV-2, the nucleotide sequence encoding the SARS-CoV-2 spike protein shown in SEQ ID NO: 1 is obtained, which is introduced into the The recombinant plasmid pTK-S was obtained from the recombinant vector of the homologous arms (J1R, J3R) of the vaccinia virus TK region, the sequence of which is shown in SEQ ID NO:2. The constructed recombinant plasmid pTK-S mainly includes vaccinia virus P7.5, P11 promoter, S and TK region homology arms (J1R, J3R).

[0026] 1.2. Construction of recombinant virus TK-S

[0027] 1.2.1. Reorganization

[0028] 293T cells (5×10 5 ) were inoculated in a six-well plate without antibiotics in complete DMEM medium, cultured overnight, and when the cell growth was 60% to ...

Embodiment 2

[0044] Example 2 Mouse Immunization Experiment of Recombinant Vaccinia Virus TK-S Expressing New Crown S Protein

[0045] 1. Method

[0046] 1.1 Mouse immunization scheme

[0047] The 6-week-old BalB / C mice were divided into 3 groups, 5 mice in each group. The first group was negative control and immunized with PBS; the second group was virus vector control and vaccinia virus dTK lacking TK was immunized; the third group was recombinant In the vaccine group, the recombinant vaccinia virus TK-S with deletion of TK and insertion of S was immunized. On the 0th day, immunization was carried out through the leg muscles. The negative control group was immunized with 100 μl PBS, and the virus vector control group was immunized with 100 μl (1×10 8 PFU / ml) dTK, the recombinant vaccine group was immunized with 100 μl (1×10 8 PFU / ml) TK-S. In the third week, the same dose and the same route were used to boost once, and blood was collected on the 12th day after each immunization for a...

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Abstract

The invention discloses a novel coronavirus vaccine based on a vaccinia virus TianTan strain. The novel coronavirus vaccine is characterized in that on the basis of the vaccinia virus TianTan strain of a vaccinia vaccine strain independently researched and developed in China, recombinant plasmids with a novel coronavirus spike protein S coding gene and a TK region homologous arm are transfected, and recombination and screening processes are performed to obtain a recombinant virus TK-S with a novel coronavirus S protein coding sequence introduced into a TK region, and expression of the novel coronavirus S protein can be detected after cells are infected by the recombinant virus. Then, a mouse is immunized with the recombinant virus, and relatively high S-specific antibody titer and neutralizing antibody titer are detected in serum of the mouse. The invention establishes the novel coronavirus vaccine based on the expression of the S protein of the vaccinia virus TianTan strain, which can solve the threat of the current new coronavirus on the global public health, and is an effective measure for treating and preventing the novel coronavirus infection.

Description

technical field [0001] The invention relates to the field of vaccine preparation, in particular to a novel coronavirus vaccine based on the Tiantan strain of vaccinia virus. Background technique [0002] Poxvirus is a large DNA virus with a genome of about 200-300kbp, mainly including Vaccinia virus, Variola virus, Cowpox virus and Monkey poxvirus Wait. Vaccinia virus has the characteristics of wide host range, large foreign gene capacity (<25kbp) and cytoplasmic replication. It is currently used in the form of vectors in the construction of various vaccines and oncolytic viruses, such as human immunodeficiency virus ( Human immunodeficiency virus (HIV) vaccine, Ebola virus (Ebola virus) vaccine, etc. Vaccinia virus Tiantan strain (VTT) is a vaccine strain independently developed by China and successfully used to prevent smallpox. It has played an important role in the process of eliminating smallpox and has been used as an emergency reserve in China to prevent smallpox...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/215A61P31/14C12R1/93
CPCC12N7/00C07K14/005A61K39/12A61P31/14A61K2039/5256C12N2770/20034C12N2770/20022C12N2710/24021
Inventor 郭斐许丰雯梅珊范张玲
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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