Anti-influenza A virus and novel universal epitope vaccine and preparing method thereof

A technology of influenza A virus and universal epitope, which is applied in the field of novel universal epitope vaccine against influenza A virus and its preparation, and can solve the problems of difficult immune response, poor immunogenicity and the like

Inactive Publication Date: 2012-07-25
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The amino acid sequence of the M2 protein is also highly conserved. This protein can stimulate the production of influenza antigens against different ge...

Method used

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  • Anti-influenza A virus and novel universal epitope vaccine and preparing method thereof
  • Anti-influenza A virus and novel universal epitope vaccine and preparing method thereof
  • Anti-influenza A virus and novel universal epitope vaccine and preparing method thereof

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Experimental program
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Embodiment 1

[0015] Influenza Virus Epitope Screening

[0016] Collect the published influenza epitopes, and download the amino acid and nucleus of H1, H3, H5, H9 subtype influenza HA gene, NP gene and M1 gene respectively from NCBI (http: / / www.ncbi.nlm.nih.gov) nucleotide sequence.

[0017] Bioinformatics MHC molecular prediction adopts: web server SYFPEITHI (http: / / www.syfpeithi.de / home.htm ), CTLpred (http: / / www.imtech.res.in / raghava / ctlpred / index.html ) , Multipre (http: / / www.i2r.a-star.Edu.sg); bioinformatics B cell epitope prediction using: web server BepiPred 1.0 sever (http: / / www.cbs.dtu.dk / services / BepiPred / ), Bcepred (http: / / www.imtech.res.in / raghava / bcepred / bcepred_submission.html), SWISS MODEL SIB Service (http: / / swissmodel.expasy.org / ) and 3D-JIGSAW Protein Comparative Modeling Server (http: / / bmm.cancerresearchuk.org / ~3djigsaw / ).

[0018] Using the method of bioinformatics, the CTL and B cell epitopes related to influenza HA, NP and M1 of various subtypes (H1, H3, H5, H9) ...

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Abstract

The invention discloses a method for preparing anti-influenza A virus and novel universal epitope vaccine. The method comprises the following steps: combining a network server and related software to predict functional epitopes of NP, M1 and HA that are related with influenza protection antigen via a bioinformatics method, obtaining a total of 8 epitopes from H1N1, H3N1, H3N2, H5N1, H5N2 subtype CTL epitope and a B-cell epitope, and adding a flexible segment GPGPG between every two of the epitopes to act as linker and naming the linked product as HMN; linking a nucleotide sequence that corresponds to the epitope polypeptide to a plasmid carrier, performing prokaryotic expression, and adding an adjuvant to the purified expressed protein to prepare the vaccine, wherein the vaccine is used to perform immunization in mice. Western blot test proves that the recombinant polypeptide is effective in gene expression and has antigenicity. The T-lymphocyte subset index of the test group is higher than that of the control group in the mice immunization test, and the result indicates that the polypeptide can induce BALC/c mice to generate specific humoral and cellular immune responses specific to the selected epitopes and proves that the polypeptide has strong immunogenicity.

Description

technical field [0001] The invention relates to a novel universal epitope vaccine against influenza A virus and a preparation method thereof. Background technique [0002] Influenza (Influenza) is caused by influenza virus (abbreviated as influenza virus, Influenza Virus, IV), an acute respiratory infectious disease that has not been effectively controlled so far. Influenza A virus has caused several large-scale epidemics all over the world. Each pandemic not only caused huge loss of human resources and material resources, but also seriously endangered the health of human beings and livestock. [0003] Avian Influenza AI (Avain Influenza AI) and Swine Influenza (Swine Influenza S1) are infectious diseases of poultry and livestock caused by influenza A virus respectively. In 1933, Smith et al first named the influenza virus isolated from human as type A influenza virus. In the 20th century, there were three plague-like global pandemics of human influenza, and the death toll...

Claims

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Application Information

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IPC IPC(8): A61K39/29A61P31/16
Inventor 许君马倩张玉娟高振兴郑文明
Owner HENAN AGRICULTURAL UNIVERSITY
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