Antigen polypeptide and application of anti-GBM nephritis model built from same

A technology of antigen polypeptide and model, applied in the field of anti-GBM nephritis model application, pathogenic antigen polypeptide field, can solve the problem of lack of human antigen highly simulated animal model of human anti-GBM nephritis, achieve high success rate, repeatability Good performance and simple preparation

Inactive Publication Date: 2016-05-18
PEKING UNIV FIRST HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem that the existing human antigen sequences in the prior art can only cause mild glomerulonephritis in rats but cannot induce the symptoms of typical Goodpasture disease, and there is also a lack of highly simulated human anti-GBM nephritis with human antigens at home and abroad The present invention provides a kind of pathogenic antigen polypeptide based on Goodpasture disease and its application in constructing anti-GBM nephritis model

Method used

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  • Antigen polypeptide and application of anti-GBM nephritis model built from same
  • Antigen polypeptide and application of anti-GBM nephritis model built from same
  • Antigen polypeptide and application of anti-GBM nephritis model built from same

Examples

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Effect test

Embodiment 1

[0044] The construction of embodiment 1 anti-GBM nephritis model

[0045] Experimental animals and materials:

[0046] WKY rats: 18 female rats, 4-6 weeks old, purchased from Beijing Weitong Lihua Company, raised at SPF level, kept at constant temperature and humidity, and illuminated for 12 hours a day;

[0047] PBS was purchased from Gino Biomedical Technology Co., Ltd., model GNM20012.

[0048] 1. Construction of antigenic polypeptide P14

[0049] The amino acid sequence of the antigenic polypeptide P14 is TDIPCPHGWISLWKGFSFIMF, which was synthesized by Beijing Zhongke Yaguang Biotechnology Co., Ltd. to obtain the antigenic polypeptide P14.

[0050] The HPLC purification results and mass spectrometry diagrams of the antigen polypeptide sequence P14 are shown in figure 1 , figure 2 , as shown in the figure, the purity of the synthesized and purified P14 meets the requirements, and the sequence is correct.

[0051] 2. Preparation of bovine α(IV)NC1 antigen polypeptide ...

Embodiment 2

[0066] Example 2 Evaluation of Kidney Damage

[0067] 1 Detection of blood creatinine level and blood urea nitrogen level

[0068] One week before rat immunization and six weeks after immunization, the rat anti-GBM nephritis model, positive control rats and negative control rats were collected from the canthus vein once a week, and the blood samples obtained each time were centrifuged separately. Treat to obtain rat serum, wherein the centrifugal force is 2000r / min, and the centrifugation time is 15min. The blood creatinine level in the rat serum was detected by the conventional alkaline picric acid colorimetric method, and the blood urea nitrogen level was detected by the conventional urease method, both of which were completed on the automatic biochemical instrument of the Laboratory Department of Peking University First Hospital. Make a standard chart of blood creatinine level according to the test results, such as image 3 As shown, and the standard chart of blood urea n...

Embodiment 3

[0078] The pathological detection of embodiment 3 rat kidney

[0079] 1 Light microscope observation of rat kidney slices

[0080] To the anti-GBM nephritis model constructed in Example 1 after 7 weeks of immunization, positive control rats and negative control rats were injected with 5% barbiturate for anesthesia, the kidneys in the rats were separated, and the capsule on the kidneys was removed , cut a small piece of renal cortex tissue from the kidney, and make a paraffin wax specimen of the kidney. The operation process is as follows:

[0081] Immediately soak the excised renal cortex tissue in neutral formalin solution to make it coagulate rapidly; then dehydrate it sequentially in 70%, 80%, 90%, 95%, and 100% concentration of alcohol, Immerse the obtained dehydrated renal cortex tissue in xylene solution to replace the alcohol to obtain a transparent renal cortex tissue, and finally immerse it in paraffin with a melting point of 60-62°C for fixation. Embed into a regul...

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Abstract

The invention discloses a pathogenic antigen polypeptide P14 of the Goodpasture disease and application of an anti-GBM nephritis model built from the same, and relates to the field of bio-medicine and medical animal models. The pathogenic antigen polypeptide is pathogenic epitope on a human Goodpasture antigen, and the amino acid sequence of the pathogenic antigen polypeptide is TDIPPCPHGWISLWKGFSFIMF. The pathogenic antigen polypeptide can be used for building the anti-GBM nephritis animal model. The animal model built through the pathogenic antigen polypeptide highly simulates human anti-GBM nephritis, the antigen is easy to prepare, obtained materials are cheap, only one-time immunity is needed, the success rate is high, repeatability is good, and the pathogenic antigen polypeptide can become a good tool for researching pathogenesis of autoimmune diseases and a good tool of intervention treatment.

Description

technical field [0001] The invention relates to a pathogenic neoantigen polypeptide and its application, in particular to a pathogenic antigen polypeptide P14 of Goodpasture disease and its application in constructing an anti-GBM nephritis model. Background technique [0002] Anti-glomerular basement membrane (glomerular basement disease, GBM) disease is a group of autoimmune disorders. The disease is characterized by circulating anti-GBM antibodies that deposit in the lungs and / or kidneys. Crescent nephritis is formed when the kidney is involved. On the one hand, the epithelial cells of the glomerulus wall layer proliferate significantly, accumulate into layers, and form crescent-shaped corpuscles around the capillary plexus. The crescent-shaped corpuscles are mainly composed of hyperplastic wall Composed of layered epithelial cells and exuded monocytes, neutrophils and lymphocytes can also infiltrate, the above components are attached to the wall of the balloon, and are d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47A61K39/00A01K67/027
CPCC07K14/47A01K67/027A61K39/0008
Inventor 崔昭赵明辉
Owner PEKING UNIV FIRST HOSPITAL
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