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Novel coronavirus compound vaccine based on DNA vector and replicating vaccinia virus vector

A vaccinia virus and virus technology, applied in the field of medical biology, can solve the problems of low protection, lack of broad-spectrum protection of mutant strains, and short duration of immune protection, and achieve the effect of preventing infection

Pending Publication Date: 2022-02-11
NAT CENT FOR AIDSSTD CONTROL & PREVENTION CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the short duration of immune protection, low protection, and lack of broad-spectrum protection against mutated strains in the existing novel coronavirus vaccine, the purpose of the present invention is to provide a DNA vector-based and replicative vaccinia virus against new coronavirus infection. The new coronavirus compound vaccine of the carrier to solve the problems existing in the existing technology

Method used

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  • Novel coronavirus compound vaccine based on DNA vector and replicating vaccinia virus vector
  • Novel coronavirus compound vaccine based on DNA vector and replicating vaccinia virus vector
  • Novel coronavirus compound vaccine based on DNA vector and replicating vaccinia virus vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: express new coronavirus S, the construction of N gene DNA vaccine

[0044] 1.1 DNA vaccine vector pDRVI3.0 and its characteristics

[0045] The DNA vaccine vector pDRVI3.0 was constructed by the Chinese Center for Disease Control and Prevention of STD and AIDS. In addition to carrying the optimally designed gene expression regulatory elements, the vector also contains a DNA sequence capable of transcribing and producing ssRNA (single-stranded RNA) with immune activation function (sequence shown in SEQ ID NO.4). The vector is disclosed in the Chinese invention patent CN101270363A, and the plasmid map is shown in figure 1 , the present invention makes it an integral part of this specification by reference.

[0046] 1.2 Target gene

[0047] The DNA vaccine expresses Sa and N antigens of 2019-nCoV. The Sa antigen is the extracellular region of the S gene of SARS-COV-2, without the transmembrane region and intracellular region, and the N antigen is the full...

Embodiment 2

[0057] Example 2: Construction and identification of recombinant vaccinia virus vaccine

[0058] The carrier used in this study is the Tiantan strain of vaccinia virus (preservation number: (CCTCC: V200416)). Vaccinia virus has the advantages of large carrier capacity, strong immunogenicity, and high safety, so we chose it as the carrier of the new crown compound vaccine.

[0059] 2.1 Synthetic genes

[0060] The antigen genes expressed by the recombinant vaccinia virus vaccine are S and N, the S gene only contains the extracellular region of the SARS-COV-2 S gene, and does not contain the transmembrane and intracellular regions, and the N antigen is the entire N of SARS-COV-2 long genes. Using the amino acid sequences of the S gene and the N gene of the new coronavirus strain NC_045512.2 as a template, it was adjusted according to the codon preference of mammalian cells and the gene was synthesized. Simultaneous gene optimization included: GC content, CpG content, mRNA seco...

Embodiment 3

[0103] Example 3: Study on Immunogenicity of DNA-Vaccinia Virus Vector Composite New Crown Vaccine

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Abstract

The invention provides a recombinant vaccinia virus capable of simultaneously expressing an extracellular segment of a new coronavirus S gene and a full-length segment of an N gene, and also provides application of an immunization kit containing the recombinant vaccinia virus, a DNA carrier capable of expressing the extracellular segment of the new coronavirus S gene and a DNA carrier capable of expressing the full-length segment of the N gene of the new coronavirus as a composite vaccine. According to the invention, experimental animals can be induced to generate specific CD4 and CD8 cell reactions aiming at new coronavirus N, S1 and S2 proteins by sequentially inoculating the three vaccines in the composite vaccine provided by the invention in 0, 2 and 4 weeks, the CD8T cell reaction is higher than the CD4T cell reaction, the induced T cell immunity can last for at least 3 months, and the combined antibody lasts for 6 months and is not obviously reduced, so that tt is shown that the composite vaccine can induce lasting immune response to SARS-CoV-2, and a neutralizing antibody induced by the vaccine has cross reaction activity and can prevent infection of various variants of the new coronavirus to a certain extent.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a DNA-vaccinia virus vector composite novel coronavirus vaccine, which can induce high-level humoral and cellular immune responses against novel coronavirus and prevent novel coronavirus infection. Background technique [0002] The pathogen of new coronary pneumonia is novel coronavirus-2 (SARS-CoV-2), which is also referred to as new coronavirus for short in the present invention. SARS-CoV-2 belongs to the β-coronavirus genus of the Coronaviridae family. It is a type of enveloped single-stranded positive-sense RNA virus with a high mutation rate, and its genome length is about 30kb. The first 2 / 3 of the genome is the non-structural gene ORF1a / b, which mainly encodes enzymes related to viral replication (RNA-dependent RNA polymerase, RdRp), and the last 1 / 3 encodes four structural proteins in turn: spike protein (S), Envelope (E), Membrane (M) and Nucleocapsid (N). Since th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/50A61K39/215A61P31/14
CPCC12N7/00C07K14/005A61K39/12A61P31/14C12N2710/24021C12N2770/20022A61K2039/5156A61K2039/53C12N2770/20034A61K2039/543A61K2039/54
Inventor 邵一鸣王书晖郝彦玲刘颖李丹任莉冯毅
Owner NAT CENT FOR AIDSSTD CONTROL & PREVENTION CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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