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Application of il-2 protein in preparation of animal vaccine adjuvant

An IL-2, 1. IL-2 technology, applied in the field of aquaculture, can solve problems such as weak immune response

Active Publication Date: 2020-07-07
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immune response elicited by oral antigen delivery alone is often weak

Method used

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  • Application of il-2 protein in preparation of animal vaccine adjuvant
  • Application of il-2 protein in preparation of animal vaccine adjuvant
  • Application of il-2 protein in preparation of animal vaccine adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, preparation of adjuvant and identification of its biological activity

[0060] 1. Cloning of rainbow trout IL-2 gene

[0061] According to the nucleotide sequence of rainbow trout IL-2 gene (GenBank: NM_001164065.1), the primer F1: 5'- GGATCC AACCCAATTCCCAGACTCCT-3' (single underline is the recognition site of restriction endonuclease BamHI), primer R1: 5'- AAGCTT TTATGAACTTAGACGCTTTGC-3' (single underline is the recognition site of restriction endonuclease HindⅢ), primer F2: 5'- GAATTC ATGAACCCAATTCCCAGACTCCT-3' (single underline is the recognition site of restriction endonuclease EcoRI, double underline is kozak sequence) and primer R2: 5'- CTCGAG TCATGAACTTAGACGCTTTGC-3' (single underline is the recognition site of restriction endonuclease XhoI).

[0062] 1. Execute the rainbow trout, quickly remove the spleen on ice aseptically, then add an appropriate amount of MEM medium containing 10% (v / v) FBS, grind the inner column of the syringe, filter...

Embodiment 2

[0127] The application of the adjuvant prepared in embodiment 2, embodiment 1

[0128] 1. Preparation of DNA vaccine (recombinant plasmid pEE12.4-G)

[0129] 1. Artificially synthesize the double-stranded DNA molecule shown in sequence 5 in the sequence listing and use it as a template, using primer pair C (by primer F3: 5'- GAATTC ATGGACACCATGATCACCACTCCG-3' (single underline is the recognition site of restriction endonuclease EcoRI) and primer R3: 5'- GGATCC TCAGGACCGGTTTGCCAGGTGAT-3' (single underline is the recognition site of the restriction endonuclease BamHI) to perform PCR amplification to obtain the PCR amplification product C of 1539bp.

[0130] 2. Take the PCR amplification product C, perform double digestion with restriction endonucleases EcoRI and BamHI, and recover the digested product C of about 1539bp.

[0131] 3. Take the pEE12.4 vector, perform double digestion with restriction endonucleases EcoRI and BamHI, and recover about 6.9 kb of vector backbone C. ...

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Abstract

The invention discloses an application of IL-2 protein to preparation of an animal vaccine adjuvant. The amino acid sequence of the IL-2 protein is shown as sequence 2 in a sequence table. Experiments prove that compared with Oncorhynchus mykiss injected with IHNV (infectious hematopoietic necrosis virus) G (glycoprotein), the level of expression of IgT gene, Mx gene, Viperin gene, CD4 gene, CD8 gene and TNF-alpha gene of Oncorhynchus mykiss injected with the IHNV G and the IL-2 protein is increased substantially, the titer of a neutralizing antibody is increased remarkably, proliferation of lymphocytes is enhanced greatly, and the protective efficacy of the IHNV G is improved effectively. Therefore, the IL-2 protein has important application value in preparation of the animal vaccine adjuvant.

Description

technical field [0001] The invention relates to the field of breeding, in particular to the application of IL-2 protein in the preparation of animal vaccine adjuvant. Background technique [0002] Fish and aquatic products are still an important source of food and nutrition for humans all over the world. At present, rainbow trout is an important component of commercial cold-water fish in most countries. However, with the increase of breeding density and the deterioration of the breeding environment, rainbow trout farming is often threatened by infectious diseases. Vaccine has been proved to be an effective means to prevent and control pathogenic microorganisms in rainbow trout. [0003] Many countries restrict the use of live vaccines due to biosafety concerns, while inactivated or genetically engineered vaccines do not provide the expected protective effect in some cases. In addition, injection immunization is currently the main method of fish vaccination, which is not on...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/205A61K48/00A61P31/14
CPCA61K39/12A61K39/39A61K2039/552A61K2039/55533C12N2760/20034
Inventor 曹永生卢彤岩李绍戊徐黎明王荻赵景壮刘红柏
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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