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T cell antigen epitope polypeptide of avian infectious bronchitis virus N protein and application of T cell antigen epitope polypeptide

A bronchitis and epitope peptide technology, applied in the directions of viral antigen components, antiviral immunoglobulins, viral peptides, etc., can solve the problems of poor ability to induce cellular immunity, clinical prevention and control obstacles, and poor cross-protection, etc. The effect of improving cross-protection, good specificity and high safety

Active Publication Date: 2021-07-13
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, clinically used IBV vaccines are mainly two types of attenuated live vaccine and inactivated vaccine. Live vaccines tend to induce mainly humoral immunity and are less capable of inducing cellular immunity
There are many serotypes of IBV, and the cross-protection between different serotype strains is poor, which brings great obstacles to the clinical prevention and control of the disease. Therefore, it is imminent to develop an IBV vaccine with cross-protection

Method used

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  • T cell antigen epitope polypeptide of avian infectious bronchitis virus N protein and application of T cell antigen epitope polypeptide
  • T cell antigen epitope polypeptide of avian infectious bronchitis virus N protein and application of T cell antigen epitope polypeptide
  • T cell antigen epitope polypeptide of avian infectious bronchitis virus N protein and application of T cell antigen epitope polypeptide

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Embodiment 1

[0031] Example 1 Screening of the T cell antigenic epitope of the N protein of avian infectious bronchitis virus

[0032] 1. Synthesis of N protein peptide library

[0033] Referring to the N protein amino acid sequence of the IBV SD strain (KY421673) published by GenBank, design a series of overlapping peptides covering the full-length amino acid sequence, each peptide is 20AA in length, and the overlapping between adjacent peptides is 10AA, a total of 40 peptides , synthesized by GenScript Biotechnology Company. Simultaneously, a non-IBV protein peptide (HA protein polypeptide of avian influenza virus H5N8, sequence: WTILKPSDTINFESN) was synthesized as an irrelevant control. The purity of the synthetic peptide is ≥95%, dissolved in DMSO, and stored at -80°C.

[0034] 2. Chicken flock immunity and isolation of spleen lymphocytes

[0035] 14-day-old SPF chickens (purchased from Beijing Boehringer Ingelheim Weitong Biotechnology Co., Ltd.) were divided into an immunization g...

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Abstract

The invention provides T cell antigen epitope polypeptide of avian infectious bronchitis virus N protein and application of the T cell antigen epitope polypeptide, and belongs to the technical field of epitope polypeptide vaccine preparation. A series of overlapped peptide fragments covering a full-length amino acid sequence are designed according to an N protein amino acid sequence of IBV, lymphocytes are respectively stimulated by the overlapped peptide fragments, the IFN-gamma secretion level of IBV specific T cells after being stimulated by the polypeptide is detected, and functional T cell epitope polypeptide is preliminarily screened out. The functional T cell epitope polypeptide is used for respectively stimulating and immunizing chicken spleen lymphocytes, and a peptide fragment capable of effectively inducing CD8 + T lymphocytes to secrete IFN-gamma is obtained through an IFN-gamma intracellular cytokine staining test. The obtained antigen epitope polypeptide has strong immunogenicity, can be used for preparing IBV multi-epitope vaccines, is high in safety and good in specificity, and has potential application value in IBV prevention and control work.

Description

technical field [0001] The invention belongs to the technical field of polypeptide vaccine preparation, and in particular relates to a T cell antigen epitope polypeptide of N protein of avian infectious bronchitis virus and application thereof. Background technique [0002] Avian infectious bronchitis (IB) is an acute, highly contagious disease caused by avian infectious bronchitis virus (IBV). The disease can occur in chickens of all ages, and the clinical symptoms are the most serious after infection in young chickens, mainly showing symptoms such as head shaking, coughing and dyspnea. Infection with some kidney-type IBV strains can also cause severe nephritis in chickens. IBV spreads quickly and spreads widely, causing huge economic losses to the world's poultry industry. IBV is a single-stranded, positive-strand RNA virus with a genome length of about 27.6kb, encoding four main structural proteins, namely spike protein (S), membrane protein (M), nucleocapsid protein (N)...

Claims

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Application Information

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IPC IPC(8): C07K14/165C12N15/50C07K16/10G01N33/569A61K39/215A61P31/14
CPCC07K14/005C07K16/10G01N33/56983A61K39/12A61P31/14C12N2770/20022G01N2333/165C12N2770/20034
Inventor 张国中覃一峰赵静赵烨杨慧明
Owner CHINA AGRI UNIV
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