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Fabricius bursa undecapeptide capable of promoting immunity

A bursal and functional technology, applied in the field of immunology, can solve the problems of bursal active peptide blank, superficial bursal active peptide, etc., and achieve the effect of simple method, low cost, and no side effects

Active Publication Date: 2015-08-12
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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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  • Fabricius bursa undecapeptide capable of promoting immunity
  • Fabricius bursa undecapeptide capable of promoting immunity
  • Fabricius bursa undecapeptide capable of promoting immunity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, the separation and identification of bursalin BP-IV:

[0033] The extraction and identification method steps of BP-IV:

[0034] ①Sample processing, take 4Kg of healthy 4-6 week old chicken bursa, crush it with 6L normal saline, mix well, freeze and thaw three times

[0035] ② Centrifuge at 4°C, 12000g, 60min, take the supernatant

[0036] ③Repeat step②

[0037] ④Ultrafiltration, 1000Da ultrafiltration membrane, freeze-dried filtrate

[0038]⑤ The sample was diluted with PBS, centrifuged at 12000g for 10min, and then passed through a 0.22um filter membrane

[0039] ⑥ Molecular sieve (Sephadex G-50) purification analysis, harvest activity peak I (see figure 1 ), and lyophilized

[0040] ⑦ Dilute solution B (2% acetonitrile, 0.065% trifluoroacetic acid), centrifuge at 12000g for 10min, and then filter with a 0.22um filter membrane.

[0041] ⑧High-performance liquid phase purification analysis Take the filtrate and reverse it to harvest the activity peak...

Embodiment 2

[0043] Embodiment 2, B cell differentiation test

[0044] 1. CFU-pre-B formation experiment:

[0045] ① Get 1×10 6 Cells / ml bone marrow mononuclear cells (select C57 / BL6 mice), add IL-7 (10ng / ml), and distribute to 6-well cell plates

[0046] ② The test group was stimulated with BP-IV (1g / ml-40g / ml) for 7 days, and the control group was stimulated with BSA (1g / ml)

[0047] ③Basis for judging the results: count the colonies with more than 40 cells

[0048] The results showed that BP-IV could significantly improve the colony formation of B cells in a dose-dependent manner ( image 3 ).

Embodiment 3

[0049] Embodiment 3, animal immunity and humoral immune response:

[0050] Studies have shown that natural bursa extract can promote the proliferation and differentiation of B cells, increase the production of antibodies, and improve the body's immunity. On this basis, the inventors used BALB / c mice (4-6 weeks old) and Laihang chickens (21 days old) as animal models, and combined immunization with gradient BP-IV twice, at 0 days and 14 days, Blood collection. The serum was separated, the level of HI antibody was detected, and the regulatory function of BP-IV on humoral immunity was analyzed (see Table 1, Table 2).

[0051] Table 1 Chicken immunization program

[0052]

[0053] Table 2 Mouse immunization program

[0054]

[0055] 1. The level of HI antibody in the chicken body experiment is detected by the hemagglutination inhibition test method. The specific operation method is:

[0056] Determine 4-fold antigen:

[0057] ①Add 25ulPBS dropwise to the 96-well V micro...

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Abstract

The invention provides Fabricius bursa undecapeptide capable of promoting the immunity. The Fabricius bursa undecapeptide is a novel polypeptide separated from Fabricius bursa tissue fluid, and the amino acid sequence of the Fabricius bursa undecapeptide is represented by the SEQ ID No.1. The provided polypeptide has an inductive effect on the generation of antibodies, and at the same time can modulate the cell factors, proliferate the lymphocyte, and promote the cellular immune response. Meanwhile, the B cell differentiation is modulated in a dose-dependent way, and the Fabricius bursa undecapeptide has a wide application prospect and can be applied to the fields such as immune regulation, immune treatment, and the like.

Description

technical field [0001] The invention relates to a bursa undecapeptide with the function of promoting immunity, which belongs to the technical field of immunology. Background technique [0002] Bioactive peptides are peptide compounds that are beneficial to the life activities of organisms or have physiological effects. Their immune regulation and antimicrobial functions play an important role in animal husbandry. [0003] The bursa of Fabricius (BF) is the central lymphoid organ of humoral immunity unique to poultry. It is a less obvious sac-like structure attached to the dorsal anal canal area of ​​the poultry cloaca. It was first discovered in 1621 by Italian anatomist Hieronymus Fabricius, hence the name. Since the bursa of Fabricius degenerates with sexual maturity, and its size is negatively correlated with the testis and adrenal glands, some researchers speculate that the BF must be some kind of endocrine gland or lymph gland related to growth and sexual development. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61K39/39A61P37/04C07K16/18
Inventor 张福波刘晓东王述柏单虎张传美李桂梅秦志华
Owner QINGDAO AGRI UNIV
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