Preparation and application of recombinant porcine interleukin-23 (IL-23) enhanced porcine circovirus 2 (PCV2) vaccine immunoadjuvant

A technology of IL-23 and P19, applied in vaccines, veterinary vaccines, recombinant DNA technology, etc., can solve the problems of less research on cytokine adjuvants, and achieve the effect of enhancing immune response and promoting growth

Inactive Publication Date: 2018-08-10
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used immune adjuvants include liposomes, chitosan, etc., but less research has been done on cytokine adjuvants

Method used

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  • Preparation and application of recombinant porcine interleukin-23 (IL-23) enhanced porcine circovirus 2 (PCV2) vaccine immunoadjuvant
  • Preparation and application of recombinant porcine interleukin-23 (IL-23) enhanced porcine circovirus 2 (PCV2) vaccine immunoadjuvant
  • Preparation and application of recombinant porcine interleukin-23 (IL-23) enhanced porcine circovirus 2 (PCV2) vaccine immunoadjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1, the construction of eukaryotic recombinant plasmid VRIL-23

[0068] The recombinant plasmid VRIL-23 is a recombinant plasmid obtained by inserting the fusion gene IL-23 into the BamHI restriction site of the VR1020 plasmid. The fusion gene IL-23 is shown in sequence 1 of the sequence listing, encoding the fusion protein IL-23 shown in sequence 2 of the sequence listing. Recombinant plasmid VRIL-23 is used to express fusion protein IL-23 in eukaryotic cells.

[0069] In sequence 2 of the sequence listing, amino acid residues 1-323 constitute the P40 fragment, amino acid residues 324-344 constitute the 2A fragment (self-cleaving connecting peptide), and amino acid residues 345-369 constitute the TPA fragment ( signal peptide), the 370-562 amino acid residues constitute the P19 fragment.

[0070] In sequence 1 of the sequence listing, nucleotides 1-969 encode the P40 segment, nucleotides 970-1032 encode the 2A segment, nucleotides 1033-1107 encode the TPA s...

Embodiment 2

[0156] Example 2, the application of eukaryotic recombinant expression plasmid VRIL-23 as a vaccine adjuvant

[0157] 1. Preparation of vaccine adjuvants

[0158] Use the ion cross-linking method to wrap the VRIL-23 plasmid, and prepare nanoparticles as a vaccine adjuvant. The specific method is:

[0159] (1) Preparation of the solution:

[0160] 10mg / ml TPP solution: Dissolve TPP in ddH 2 In O, make the TPP concentration be 10mg / ml, obtain the TPP solution of 10mg / ml;

[0161] Chitosan solution of 2.4mg / ml: Chitosan is dissolved in 1% (pH5.5) glacial acetic acid aqueous solution, makes chitosan concentration be 2.4mg / ml, obtains the chitosan solution of 2.4mg / ml ;

[0162] Filter the prepared solution with a 0.22m microporous membrane to sterilize;

[0163] (2) Premixing of plasmid and TPP solution: mix the eukaryotic recombinant plasmid VRIL-23 solution (solvent is water) with an appropriate amount of TPP solution, and incubate at 55°C for 20 minutes to obtain a premixe...

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Abstract

The invention discloses preparation and application of a recombinant porcine interleukin-23 (IL-23) enhanced porcine circovirus 2 (PCV2) vaccine immunoadjuvant. The P40 and P19 gene fragments of a Tibetan pig IL-23 gene is obtained by using a gene cloning technology for the first time, and a 2A self-shearing expression technology is used for linking the P40 and P19 genes, so that a complete IL-23gene sequence is obtained; then, an In-Fusion cloning method is used for enabling the complete IL-23 gene to be constructed into an eukaryotic secreting type expression vector VR1020; chitosan nanoparticles are prepared by wrapping IL-23 plasmid with chitosan, and experimental animals are inoculated with the chitosan nanoparticles and a PCV2 vaccine at the same time; when pigs are used as the experimental animals, the results indicate that the IL-23 eukaryotic transfection plasmid can effectively enhance the immune response of the pigs to the PCV2 vaccine and promote the growth of vaccinated animals; a new technology is provided for the development of the new and efficient PCV2 immunoadjuvant.

Description

technical field [0001] The invention relates to the preparation and application of a recombinant porcine IL-23 enhancing PCV2 vaccine immune adjuvant. Background technique [0002] Pork products are the main source of meat consumption for urban and rural residents and are closely related to people's lives. Since the 21st century, the annual number of live pigs and slaughtered pigs in my country has always accounted for about 50% of the world's total, ranking first in the world. While my country's pig industry has achieved world-renowned achievements, there are still many factors that restrict the healthy development of my country's pig industry, among which disease is the most important factor. Porcine circovirus type 2 (PCV2) infection is a relatively common type of infectious disease. PCV2 is the main pathogen that causes Postweaning multisystemic wasting syndrome (PMWS) in weaned piglets. Infection has caused significant economic losses to the swine industry worldwide....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63A61K39/39A61K39/12A61P31/20
CPCA61K39/12A61K39/39A61K2039/552A61K2039/55527A61P31/20C07K14/54C07K2319/02C12N2750/10034A61K2300/00
Inventor 高荣肖永乐方鹏飞陈祎邱文英宋婷玉李金海王泽洲吕学斌李江凌龚文波
Owner SICHUAN UNIV
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