Molecular biological significance of asialoglycoprotein receptor in autoimmune hepatitis type I

自身免疫性I型肝炎脱唾液酸糖蛋白受体的分子生物学意义

基本信息

批准号：
06454261
负责人：
KIYOSAWA Kendo
金额：
$ 4.61万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454261/
关键词：
autoimmune hepatitis asialoglycoprotein receptor autoantibody molecular biology

项目摘要

Reverse transcription and polymerase chain reaction was applied to isolate cDNAs encoding asialoglycoprotein receptor (ASGPR) H1 and L-H2 from human liver mRNAs. Using these cDNAs, recombinant ASGPR protein was expressed in E.coli. Rabbit antisera specific to human ASGPR H1 or L-H2 was obtained by immunizing these recombinant proteins. Enzyme-linked immunosorbent assay (ELISA) using bacterial recombinant antigen was developed to measure serum antibodies to ASGPR.Sera from autoimmune hepatitis, primary biliary cirrhosis, alcoholic liver disease, chronic hepatitis C,chronic hepatitis B,systemic lupus erythematosus, and healthy subjects were tested. High OD value was obtained almost exclusively in autoimmune hepatitis and chronic hepatitis C.The OD value of anti-ASGPR antibodies detected by ELISA using recombinant antigen was correlated with anti-ASGPR antibody titers that were determined by ELISA using human liver antigen. By using extracellular domain of ASGPR protein in ELISA,similar sensitivity to pick-up anti-ASGPR antibodies was demonstrated in autoimmune hepatitis and less reactivity was detected in chronic hepatitis C.It was suggested that main epitope which was reactive with antibodies in autoimmune hepatitis could be located on the extracellular domain of the ASGPR protein. Lymphocytes from peripheral blood mononuclear cells in a patient with autoimmune hepatitis showed a stimulation index as 4 in lymphocyte stimulation test with recombinant ASGPR antigen. There could exist a certain class of lymphocytes reactive with human ASGPR in autoimmune hepatitis patients.

将逆转录和聚合酶链反应用于分离人肝脏mRNA的编码Asialoglycoprotein受体（ASGPR）H1和L-H2的cDNA。使用这些CDNA，在大肠杆菌中表达重组ASGPR蛋白。通过免疫这些重组蛋白来获得针对人ASGPR H1或L-H2的兔抗血清。开发了使用细菌重组抗原的酶联免疫吸附测定法（ELISA），以测量自身免疫性肝炎的ASGPR.SERA血清抗体，原发性胆汁肝硬化，酒精性肝病，慢性肝炎，慢性肝炎，慢性肝炎，慢性肝炎，肠胃不适，睾丸效果。在自身免疫性肝炎和慢性丙型肝炎中，几乎完全获得了高OD值。使用重组抗原与ELISA检测到的抗ASGPR抗体的OD值与使用人肝抗原通过ELISA确定的抗ASGPR抗体滴度相关。 By using extracellular domain of ASGPR protein in ELISA,similar sensitivity to pick-up anti-ASGPR antibodies was demonstrated in autoimmune hepatitis and less reactivity was detected in chronic hepatitis C.It was suggested that main epitope which was reactive with antibodies in autoimmune hepatitis could be located on the extracellular domain of the ASGPR protein.自身免疫性肝炎患者的外周血单核细胞的淋巴细胞在淋巴细胞刺激试验中显示出刺激指数为4，并通过重组ASGPR抗原。在自身免疫性肝炎患者中，可能存在某种类型的淋巴细胞与人ASGPR反应。