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Polypeptide with immunomodulatory effects

A vaccine and amino acid technology, applied in blood/immune system cells, peptides, protozoan antigen components, etc., can solve the problems of blank bursa active peptide, superficial cognition of bursa active peptide, etc., and achieve good effect and immunity. The effect of very weak originality and simple structure

Inactive Publication Date: 2015-03-11
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Some progress has been made in the study of bursal active peptides, but at present, the understanding of bursal active peptides is still relatively superficial, and there are still many aspects to be resolved about their biological characteristics and functions, especially for The application of the known bursa active peptide is almost blank, further research is needed

Method used

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  • Polypeptide with immunomodulatory effects
  • Polypeptide with immunomodulatory effects
  • Polypeptide with immunomodulatory effects

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Isolation and identification of bursalin undecapeptide:

[0032] Take 4000 grams of the bursa of fabric (AA broiler, affiliated to Shanghai Academy of Agricultural Sciences) from healthy 4-6 weeks old chickens, crush them with 6000ml of normal saline, mix them, and freeze and thaw them three times. Centrifuge at 12000g for 60 minutes at 4°C, transfer the supernatant to another tube, and centrifuge again. Then pass through a 1000 Da ultrafiltration membrane, ultrafiltration, and freeze-dry the filtrate. The lyophilized sample was diluted with PBS, centrifuged at 12000g for 10 minutes, and then filtered through a 0.22um filter. The filtrate was purified and analyzed by molecular sieve (Sephadex G-50), and the activity peak I was harvested (see figure 1 ) And freeze-dried. The lyophilized sample was diluted with solution B (2% acetonitrile, 0.065% trifluoroacetic acid), centrifuged at 12000g for 10 minutes, and then filtered with a 0.22um filter membrane. The fi...

Embodiment 2

[0034] Example 2. Animal immunity and humoral immune response:

[0035] Natural bursa extract can promote the proliferation and differentiation of B cells, leading to the production of antibodies. On this basis, the 21-day-old Laihang chicken was used as an animal model, and avian influenza inactivated vaccine (H9N2 subtype F strain, purchased from Nanjing Merial Animal Health Co., Ltd.) was combined with different concentrations of Bursa Undecapeptide. Two times, 14 days after the two immunizations, blood was collected, serum was separated, changes in HI antibody levels were detected, and the regulatory function of bursa undecapeptide on humoral immunity was analyzed (see Table 1). Through this experiment, it can be found that the undecapeptide of Bursa Fabricius can significantly regulate the humoral immune function, is closely related to the dose, and has the function of balancing Th1 and Th2 immune responses.

[0036] 1. The level of HI antibody is detected by hemagglutination...

Embodiment 3

[0051] Example 3. B cell differentiation test

[0052] 1. CFU-pre-B formation experiment:

[0053] Take bone marrow mononuclear cells from C57 / BL6 mice ((1×10 6 Cells / ml)), add methylcellulose medium containing IL-7 (10ng / ml), and distribute to 6-well cell plates. It was stimulated with BP11 (1μg / ml to 40μg / ml) for 7 days, and the control group was stimulated with BSA (1μg / ml). Colonies larger than 40 cells were counted. The results show that BP11 can significantly increase the colony formation of B cells and is dose-dependent, see attached Figure 7 .

[0054] 2. B cell differentiation experiment:

[0055] Take bone marrow mononuclear cells from C57 / BL6 mice ((1×10 6 Cells / ml)), IL-7 (10ng / ml) was added, and aliquoted into 6-well cell plates. It was stimulated with BP11 (1μg / ml to 40μg / ml) for 7 days, and the control group was stimulated with BSA (1μg / ml). Add PE, FITC, PE-Cy5 and PE-Cy7 labeled CD43, anti-IgD and anti-IgM monoclonal antibodies (eBioscience, USA) for 30m at room ...

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Abstract

The invention aims to provide polypeptide with immunomodulatory effects, namely undecapeptide which has immunomodulatory effects and is separated from bursa of Fabricius. The amino acid sequence of the polypeptide is SEQ ID NO:1. The bursa of Fabricius undecapeptide separated from a bursa of Fabricius extract product has a simple structure, has no toxic or side effect, and has poor immunogenicity. The polypeptide can be obtained by separation and extraction from bursa of Fabricius and also can be obtained by chemical synthesis. The polypeptide has low cost and can be put into mass production. The polypeptide has an induction effect on antibody production, and also can regulate production of cytokines and proliferation of lymphocyte and promote cellular immune response. In addition, the polypeptide also regulates differentiation of B cell in a dose-dependent mode. The polypeptide has a wide application prospect, taking an application of the polypeptide in the aspects of immunomodulation, treatment and the like for example.

Description

Technical field [0001] The invention belongs to the technical field of functional protein / polypeptide screening, and specifically relates to a polypeptide with immunomodulatory effect. Background technique [0002] Bursa of Fabricius (BF) is a less obvious cyst-like structure attached to the dorsal anal area of ​​the cloaca of birds. The Bursa of Fabricus (BF) was first discovered in 1621 by the Italian anatomist Hieronymus Fabricius, hence the name. Initially, Fabrizius speculated that the hollow structure of the bursa of Fabricius might be used as a container for storing semen. However, this is not the case. For hundreds of years, the function of the bursa of Fabricius has plagued many researchers. Because the bursa degenerates with sexual maturity, and its size is negatively related to the testis and adrenal glands, some researchers speculate that BF must be some kind of endocrine or lymph gland related to growth and sexual development. As early as 1956, Glick and others ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61K39/002A61P37/00C12N5/0781
Inventor 刘晓东单虎陈溥言王述柏杨海燕秦志华张传美
Owner QINGDAO AGRI UNIV
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