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Specific autoantibody of PBC (primary biliary cirrhosis) and application

A technology of biliary cirrhosis and autoantibodies, applied in the field of immunology, can solve problems such as easy misdiagnosis

Inactive Publication Date: 2013-04-17
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antimitochondrial autoantibodies (AMA) is the main diagnostic indicator of the disease, but there are still some patients who are negative for this antibody, which is easy to be misdiagnosed

Method used

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  • Specific autoantibody of PBC (primary biliary cirrhosis) and application
  • Specific autoantibody of PBC (primary biliary cirrhosis) and application
  • Specific autoantibody of PBC (primary biliary cirrhosis) and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Detection of Sp20 autoantibodies in PBC patients

[0020] 1. Case selection

[0021] 1. Thirty-five PBC patients were inpatients from Department of Hepatology, Peking University People's Hospital from 1995 to 2004, including 34 females and 1 male, aged 34-77 years, with an average of (54±10) years old. The control group included 8 patients with chronic hepatitis B, 5 patients with chronic hepatitis C, 20 patients with Sjogren's syndrome, 8 patients with autoimmune hepatitis, and 9 healthy blood donors. The serum of patients with Sjogren's syndrome was provided by the Immunology Laboratory of Peking University People's Hospital, the serum of healthy blood donors was provided by the blood bank of Peking University People's Hospital, and the serum of patients with primary biliary cirrhosis, chronic hepatitis B, chronic hepatitis C and autoimmune hepatitis Provided by the Department of Hepatology, all serum used in the research was stored at -20°C.

[0022] 2. D...

Embodiment 2

[0075] Example 2 Subcellular localization distribution and preliminary identification of Sp20 protein

[0076] (1) Materials and methods

[0077] 1. Serum: From the sera of 8 anti-Sp20 positive PBC patients, select a patient serum containing anti-PDC-E2 and anti-Sp20.

[0078] 2. Cell lines

[0079]Human liver cancer cell line HepG2.2.15, human cervical cancer cell line HeLa, human breast cancer cell line Mcf-7, human melanoma cell line Mel-ed1, human poorly differentiated gastric adenocarcinoma cell line BCG-823, human small cell lung cancer cell line NC1-H446, rat liver cell line BRL and human cholangiocarcinoma cell line RBE were purchased from the Surgical Laboratory of Peking University People's Hospital. The degree of cell differentiation was obtained from the original literature of strain establishment; RBE, Mcf-7, and BRL were cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (PAA); the other five types of cells were cultured in 10% % fetal bovi...

Embodiment 3

[0104] Example 3 Application of Sp20 Antibody in Preparation of Kit for Diagnosis of Primary Biliary Cirrhosis

[0105] By studying the mechanism of Sp20 antibody in the pathogenesis of PBC, it provides a theoretical basis for early diagnosis, early treatment and prognosis of PBC patients. It is helpful for the diagnosis of AMA-negative PBC patients and for the differential diagnosis of other autoimmune diseases.

[0106] The Sp20 antibody can be used in the preparation of a kit for diagnosing primary biliary cirrhosis.

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Abstract

The invention provides a specific autoantibody of the PBC and a corresponding antigen. RBE (relative biological effectiveness) TP (total protein) of cholangiocarcinoma cells is taken as the antigen of the specific autoantibody; an IB (immunoblotting) method is adopted to detect the serum of a PBC patient; an autoantibody aiming at protein with the molecular weight about 20kD is detected and named Sp20 antibody; and then the Sp20 antibody is separated and obtained from the serum of the PBC patient through two-dimensional electrophoresis. The invention further provides an application of the Sp20 antibody in preparation of a kit for PBC diagnosis. By means of research on the mechanism of the Sp20 antibody in PBC incidence, theoretical basis is provided for early diagnosis, early treatment and prognostic judgment of PBC patients, diagnosis for AMA (anti mitochondrial antibody)-negative PBC patients is facilitated, and differential diagnosis with other autoimmune diseases is facilitated.

Description

technical field [0001] The present invention relates to the field of immunology, in particular to a specific autoantibody of primary biliary liver cirrhosis, its corresponding antigen and their application. Background technique [0002] Primary biliary cirrhosis (PBC) is a chronic, progressive cholestatic liver disease that is generally believed to be related to autoimmunity. The lesion is characterized by non-suppurative inflammation of the small intrahepatic bile ducts. PBC is more common in middle-aged and elderly women, with a male to female ratio of 1:8. At present, there is no cure for the disease, but early and timely treatment can improve biochemical indicators and liver histological lesions, thereby delaying the progression of the disease. Once it enters the advanced stage, liver transplantation is the only feasible treatment, so early diagnosis and early treatment are very important . However, due to the atypical clinical manifestations of the disease in the ear...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K14/47G01N33/68
Inventor 魏来张海莹王江华
Owner PEOPLES HOSPITAL PEKING UNIV
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