A kind of active peptide and application of promoting the immune effect of h9n2 avian influenza vaccine

A technology of immune effects and active peptides, applied in the field of active peptides, can solve the problems of influenza viruses that are more difficult and cannot cope with different virus subtypes

Active Publication Date: 2019-07-19
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the rapid mutation, the vaccines currently used in clinical practice can no longer cope with different virus subtypes, and it is even more difficult to completely prevent and control i

Method used

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  • A kind of active peptide and application of promoting the immune effect of h9n2 avian influenza vaccine
  • A kind of active peptide and application of promoting the immune effect of h9n2 avian influenza vaccine
  • A kind of active peptide and application of promoting the immune effect of h9n2 avian influenza vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Isolation and identification of BP4

[0026] Take 500 grams of the bursa of a healthy 4-6 week-old chicken (AA broiler, farm affiliated to Shanghai Academy of Agricultural Sciences), crush it with 1000 ml of normal saline, freeze and thaw three times, and centrifuge at 14000 g for 60 minutes at low temperature. The supernatant was ultrafiltered at 1000 Da and freeze-dried. After the lyophilized sample was diluted, it was filtered with a 0.22um filter membrane, and then the reverse high performance liquid phase purification analysis was performed, and the activity peak with a retention time of 15.72min was harvested (see figure 1 ). The harvested eluate was analyzed by MALDI-TOF-MS, the molecular weight was 431.213, the amino acid sequence GERA (see figure 2 ).

Embodiment 2

[0028] 1. Synthetic BP4

[0029] Entrust commercial peptide synthesis company according to COOH-GERA-NH2 amino acid

[0030] The sequence (SEQ ID NO: 1) synthesizes the polypeptide, and the purity is required to be ≧97%.

[0031] 2. Vaccines

[0032] Inactivated bird flu vaccine (H9N2 subtype F strain, purchased from Nanjing Meria Animal Health Co., Ltd.)

[0033] 3. Grouping of experimental animals

[0034] 50 BALB / C mice were randomly divided into five groups, 10 in each group: (1) avian influenza inactivated vaccine immunization group (0.2ml of each immune inactivated vaccine); (2~4) inactivated vaccine+BP4 group (BP4 concentrations were 10, 50, 250ug / mL, and the immunization dose was 0.2ml); PBS control group (each immunized with 0.2ml PBS), immunized twice with an interval of 2 weeks.

[0035] 4. Sample Collection

[0036] Two weeks after the second immunization, blood was collected from the orbit every week, and the serum was separated by centrifugation at 8,000×g f...

Embodiment 3

[0048] 1. Synthetic BP4

[0049] Entrust a commercial peptide synthesis company according to COOH-GERA-NH2 amino acid, the purity is ≧97%.

[0050] 2. Vaccines

[0051] Inactivated bird flu vaccine (H9N2 subtype F strain, purchased from Nanjing Meria Animal Health Co., Ltd.)

[0052] 3. Grouping of experimental animals

[0053]45 75-day-old chickens (Baihang chickens, purchased from Qinglongshan Farm) were randomly divided into three groups, 15 in each group: (1) Bird flu inactivated vaccine immunization group (each immunized inactivated vaccine 0.5ml) (2) Inactivated vaccine+BP4 group (BP4 concentration is 50ug / mL, immunization dose is 0.5ml); PBS control group (each immunized with 0.5ml PBS).

[0054] 4. Sample Collection

[0055] On the 2nd and 4th weeks after immunization, anticoagulant blood was collected from each chicken wing, and the serum was separated by centrifugation at 8000×g for 10 minutes. Antibody titers and cytokine expression levels were measured, and the...

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Abstract

The invention relates to active peptide for promoting H9N2 avian influenza vaccine and application. The immunity active peptide is small-molecular polypeptide from bursa of Fabricius; the amino acid composition is Gly Glu Arg Ala-NH2; the structure is simple; the immunogenicity is weak. The active peptide has a wide immune enhancement effect; the effects of irritating antibody generation, regulating the cell factor and improving the vaccine immunity efficacy are achieved on mouse and poultry. The active peptide can be used as an adjuvant or an immunopotentiator to be applied to animal vaccine, so that the animal disease resistance capability and the specific antigen immune response capability are improved; the vaccine immune efficiency is improved; the active peptide can be applied to the fields of basic immune research, clinical application research and the like.

Description

technical field [0001] The invention relates to an active peptide of an H9N2 bird flu vaccine, belonging to the technical field of veterinary biological products. Background technique [0002] Bird flu causes major economic losses and disastrous consequences every year. New recombinant virus strains emerge in an endless stream. Influenza viruses have broken through the interspecies barrier, and the range of infected hosts continues to expand. Since the 20th century, the epidemic of influenza has deprived the lives of tens of millions of people and caused a huge threat to the public health of all mankind. In view of the rapid mutation, the vaccines currently used in clinical practice can no longer cope with different virus subtypes, and it is even more difficult to completely prevent and control influenza viruses. Therefore, how to induce the body to produce an effective and durable immune protection response is an urgent need for relevant scientific research staff to solve ...

Claims

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Application Information

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IPC IPC(8): C07K14/465C07K5/103A61K39/145A61K39/39A61P31/16
CPCA61K39/12A61K39/39A61K2039/552A61K2039/55516C07K5/1008C07K14/465C12N2760/16034
Inventor 冯秀丽宗嫚嫚周川杰郑阳余远楠曹瑞兵陈溥言
Owner NANJING AGRICULTURAL UNIVERSITY
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