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Method for extracorporeally preparing transfer factor against specificity of infectious bronchitis virus of chickens

A technology for bronchitis and chicken infectivity, applied in antiviral agents, medical formulas, respiratory diseases, etc., can solve the problems of non-specificity of diseases, unstable treatment effect, etc., and achieve the effect of improving immunity

Inactive Publication Date: 2010-06-30
TIANJIN SHENGJI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the disadvantages of the transfer factor prepared in the prior art that have no specificity for diseases and unstable therapeutic effect, and provide an in vitro immune method with strong production specificity, low cost, high output rate and better effect. Preparation method of anti-chicken infectious bronchitis virus specific transfer factor

Method used

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  • Method for extracorporeally preparing transfer factor against specificity of infectious bronchitis virus of chickens
  • Method for extracorporeally preparing transfer factor against specificity of infectious bronchitis virus of chickens

Examples

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Effect test

Embodiment 1

[0016] A preparation method for preparing anti-chicken infectious bronchitis virus-specific transfer factor in vitro, the steps are:

[0017] (1) Aseptically take anticoagulant blood, and prepare a monolayer of lymphocytes with peripheral blood, the steps are:

[0018] Take 25 ml of animal whole blood through a sterile vein, add 0.3 ml of 0.8% heparin solution for anticoagulation, let it stand for 45 minutes, and use a pipette with a rubber ball to absorb all the plasma above the red blood cell layer, especially the white blood cell layer immediately above the red blood cell layer. Aspirated, divided into sterilized centrifuge tubes, washed repeatedly 2 to 3 times with PBS (pH 7.2) solution, centrifuged at 800 to 1200 rpm, and then cultured white blood cells with 40 ml of hydrolyzed milk protein containing 30% calf serum. After uniform mixing, it is divided into 100 ml culture square bottles or brick bottles, the bottle stopper is tightly closed, and cultured in a 37°C incubat...

Embodiment 2

[0025] A preparation method for preparing anti-chicken infectious bronchitis virus-specific transfer factor in vitro, the steps are:

[0026] (1) Aseptically take the spleen of the experimental chicken, and use the spleen to prepare a monolayer of lymphocytes, the steps are:

[0027] A. Processing chicken spleen

[0028] Take fresh chicken spleen and put it in a sterile glass container, add PBS (pH 7.2) solution to soak for half an hour or immerse in 1% bromogeramine solution by mass percentage, take out the spleen surface with alcohol disinfection, use dissecting scissors and tweezers Remove the spleen fat and capsule, wash once with PBS (pH 7.2), then cut the spleen into small pieces and transfer to a plate, and finally cut the spleen into 1mm3 small pieces with ophthalmic scissors, and then wash with PBS (pH 7.2) 2-3 times to remove blood cells, pigment substances and cells damaged mechanically during the shredding process;

[0029] B. Digest and disperse tissue blocks

...

Embodiment 3

[0047] A preparation method for preparing anti-chicken infectious bronchitis virus-specific transfer factor in vitro, the steps are:

[0048] (1) Take fresh chicken spleen and place it in a sterile glass container, add PBS (pH 7.2) solution to soak for half an hour or immerse in brogeramine solution with a mass percent concentration of 1%, take out the surface of the spleen and disinfect it with alcohol, and dissect it Scissors and tweezers remove the spleen fat and capsule, wash once with PBS (pH 7.2), then cut the spleen into small pieces and transfer them to a plate, and finally cut the spleen into 1 mm ) liquid washing 2-3 times;

[0049] (2) Suck off the PBS (pH7.2) solution washed in the previous step, move the shredded tissue pieces into an Erlenmeyer flask, and add 0.25% trypsin solution (pH 7.6~ 7.8), placed in a 37°C water bath for 20-40 minutes for digestion. Shake the Erlenmeyer flask every 10 minutes to disperse the tissue pieces and continue digestion until the...

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Abstract

The invention relates to a method for extracorporeally preparing a transfer factor against the specificity of infectious bronchitis viruses of chickens. In the invention, splenic organs or peripheral blood of chickens are used as raw materials to make a lymphocyte monolayer, and then phytohemagglutinin and infectious bronchitis viruses of chickens are used to culture lymphocytes through induction so as to extracorporeally produce a large quantity of transfer factors against the specificity of the infectious bronchitis viruses of chickens. The anti-specificity transfer factor prepared by using the method of the invention is used for treating infectious bronchitis of chickens and can raise the immunity of poultry.

Description

technical field [0001] The invention relates to a method for preparing anti-chicken infectious bronchitis virus-specific transfer factor in vitro. Background technique [0002] Transfer factor is a low-molecular-weight polypeptide and nucleic acid complex extracted from lymphocytes, which can specifically or non-specifically improve the immune function of the body and transmit immune information. It is known as a cellular immune activator. It has no species specificity, can be transmitted between species, and animal sources can be used in humans. [0003] The routine preparation method of transfer factor is: remove the fat and connective tissue from the fresh or frozen spleen and lymph nodes of animals at about 25°C, and then wash them with purified water; cut the washed tissues into small pieces, add appropriate amount of water, and use tissue Break the cells with a masher, make a homogenate, add an appropriate amount of purified water, mix well, freeze and thaw repeatedly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/14A61K35/28A61P11/00A61P31/14A61K35/57
Inventor 田杏芳王连民
Owner TIANJIN SHENGJI GRP CO LTD
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