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Method for detecting content of soluble CD28 in blood of patients suffering Graves disease

A soluble, disease-free technology, applied in the medical field, can solve problems such as unclear

Inactive Publication Date: 2012-10-17
SUZHOU HEALTH COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the clinical diagnostic value of soluble CD28 molecule in Graves' disease and what role it plays in the immune response and immune regulation in the body will play an important role in the occurrence, development and outcome of diseases such as tumors, immunodeficiency diseases, autoimmune diseases and organ transplantation. What is the meaning of it is still unclear

Method used

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Embodiment 1

[0032] 1. Main materials and reagents

[0033] Bovine serum (Hyclone, U.S.); full medium: add 100 ml of fetal bovine or calf serum, 0.15 g of L-glutamine, and NaHCO 3 2.0g, sodium pyruvate 0.11g, glucose 3.6g, HEPES 4.766g, 2-mercaptoethanol 10.0ml (5×10 -3 mol / L); CD28 monoclonal antibodies 2D5, 2F5, 3B6, 3F8 and 8G8 (developed by our department); affinity chromatography column: Protein G immunoaffinity chromatography column (Pharmacia, Sweden); rhCD28 / Fc recombinant protein ( R&D, USA); succinylhydroxybiotin (Sigma, USA); Dimethylsulfoxide (Sigma, USA); 4% polylysine (Sigma, USA); streptavidin-HRP (Roche, Switzerland); avidin -PE (Immunotech, France); Enzyme label assay plate (8 × 12 holes, Costar, U.S.); Enzyme label assay instrument (Bio-Rad, U.S.); Culture bottle: 50ml, 500ml plastic culture bottle (Nunc, Denmark); Experimental instrument: CO 2 Incubator, centrifuge (Jouan, France); inverted microscope and fluorescence microscope (Olympus, Japan); flow cytometer (Beckm...

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Abstract

The invention discloses a method for detecting the content of soluble CD28 in blood of patients suffering Graves disease. The method comprises: preparing materials, reagents and blood specimens, then sequentially conducting biotin labeling of a monoclonal antibody, identification of the biotin-labeled monoclonal antibody, drawing of an sCD28 standard work curve, separation and purification of T cells in Graves-suffering patient blood, phenotypic analysis of the T cells of the Graves-suffering patients, determination of the content of soluble CD28 in the Graves-suffering patient blood, immunoblotting analysis of the soluble CD28 in the patient blood, determination of the influence of the soluble CD28 on cytokines secreted by dendritic cells, determination of the influence of the soluble CD28 on dendritic cell NF-kB nuclear translocation and other determination steps, as well as statistical analysis. The method for detecting the content of soluble CD28 in blood of patients suffering Graves disease in the invention provides an experimental basis and theoretical base for further investigating the establishment of an optimized in vitro detection scheme by combining other soluble costimulatory molecules and the promotion significance in clinical diagnosis, treatment and prognosis assessment of autoimmune diseases, and for searching new immunologic intervention means.

Description

technical field [0001] The invention relates to the medical field, in particular to a method for measuring the content of soluble CD28 in the blood of Graves disease patients. Background technique [0002] Graves disease, also known as diffuse goiter with hyperthyroidism (hyperthyroidism) or toxic diffuse goiter, is the most common type of hyperthyroidism. It is also an organ-specific autoimmune disease with increased secretion of thyroid hormone, and its etiology and pathogenesis are still unclear. At present, it is believed that its immunopathogenic mechanism is due to the thyroid-stimulating hormone (TSH) receptor on the surface of thyroid cells acting as an antigen to stimulate the body to produce antibodies (TRAb) that mimic the role of TSH and bind to TSH receptors to continuously stimulate thyroid follicular cells. , Formation and secretion of excess thyroxine and cause hyperthyroidism. In recent years, people have paid more and more attention to the role of immune ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/68G01N33/532G01N33/531C12Q1/02
Inventor 孙中文
Owner SUZHOU HEALTH COLLEGE
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