To better understand the effect of a new split variant of human asialoglycoprotein receptor (ASGPR H1b) on ASGPR ligands’ binding ability, we established a functional cell line which expresses ASGPR.The full lengths ...To better understand the effect of a new split variant of human asialoglycoprotein receptor (ASGPR H1b) on ASGPR ligands’ binding ability, we established a functional cell line which expresses ASGPR.The full lengths of ASGPRH1a and H2c fragments from human liver were amplified by reverse transcript PCR (RT-PCR) and inserted into eukaryotic expression vector pIRES2EFP, pCDNA3.1 (Zeo+) respectively.The recombinants were cotransfected into HeLa cells.After selection by using Neocin and Zeocin, a stably transfected cell line was established, which was designated 4-1-6.The transcription and expression of ASGPRH1a and H2c in 4-1-6 were confirmed by RT-PCR, Western blotting and immunofluorescence.The endocytosis function of the artificial "ASGPR" on the surface of 4-1-6 was tested by FACS.It was found that the cell line 4-1-6 could bind ASGPR natural ligand molecular asialo-orosomucoid (ASOR).After the eukaryotic plasmid H1b/pCDNA3.1 (neo) was transfected into cell line 4-1-6, H1b did not down-regulate the ligand binding ability of ASGPR.The eukaryotic expression plasmid H1b/pcDNA3.1 (neo) and H2c/pcDNA3.1 (neo) were co-transfected transiently into Hela cell.Neither single H1b nor H1b and H2c could bind ASOR.In conclusion, a functional cell line of human asialoglycoprotein receptor (ASGPR) which expresses both H1a and H2c stably was established.The new split variant H1b has no effect on ASGPR binding to ASOR.ASGPRH1b alone can’t bind to ASOR, it yet can’t form functional complex with ASGPRH2c.展开更多
目的:分析乙型肝炎病毒(hepantitis B virus,HBV)和丙型肝炎病毒(hepantitis C virus,HCV)感染患者血清中肝特异性抗体水平,探讨其与肝细胞损伤的关系。方法:收集2011年1月至2011年10月川北医学院附属医院HBV和HCV感染患者和健康体检者...目的:分析乙型肝炎病毒(hepantitis B virus,HBV)和丙型肝炎病毒(hepantitis C virus,HCV)感染患者血清中肝特异性抗体水平,探讨其与肝细胞损伤的关系。方法:收集2011年1月至2011年10月川北医学院附属医院HBV和HCV感染患者和健康体检者血清,检测患者丙氨酸氨基转移酶(alannine aminotransferase,ALT)和总胆红素水平;通过酶联免疫吸附法检测抗肝细胞膜特异性脂蛋白(liver specific lipoprotein,LSP)和抗去唾液酸糖蛋白受体(asialoglycoprotin receptor,ASGPR)的血清水平;以ALT>40分为肝功能异常组与正常组。使用SPSS13.0统计软件分析结果,P<0.05认为有统计学意义。结果:HBV(H)与健康对照组比较anti-LSP、anti-ASGPR具有统计学差异,HBV组患者anti-ASGPR水平与ALT具有相关性。HCV(H)与健康对照组比较anti-LSP具有统计学差异,HCV组患者anti-LSP与anti-ASGPR具有相关性。HBV患者anti-LSP、anti-ASGPR表达水平较HCV患者高。结论:HBV、HCV感染后,anti-LSP和anti-ASGPR表达高于健康对照组,ALT水平升高时抗肝特异性抗体表达明显增高,表明其可能作为监测肝细胞损伤的特异性免疫学指标。展开更多
基金supported by grants from the National Major Science and Technology Special Project for Infectious Diseases of China (No.2008ZX10002-011)National High Technology Research and Development of China (Program 863) (No.2006AA02Z128)the National Natural Science Foundation of China (Nos.30700701,30571646)
文摘To better understand the effect of a new split variant of human asialoglycoprotein receptor (ASGPR H1b) on ASGPR ligands’ binding ability, we established a functional cell line which expresses ASGPR.The full lengths of ASGPRH1a and H2c fragments from human liver were amplified by reverse transcript PCR (RT-PCR) and inserted into eukaryotic expression vector pIRES2EFP, pCDNA3.1 (Zeo+) respectively.The recombinants were cotransfected into HeLa cells.After selection by using Neocin and Zeocin, a stably transfected cell line was established, which was designated 4-1-6.The transcription and expression of ASGPRH1a and H2c in 4-1-6 were confirmed by RT-PCR, Western blotting and immunofluorescence.The endocytosis function of the artificial "ASGPR" on the surface of 4-1-6 was tested by FACS.It was found that the cell line 4-1-6 could bind ASGPR natural ligand molecular asialo-orosomucoid (ASOR).After the eukaryotic plasmid H1b/pCDNA3.1 (neo) was transfected into cell line 4-1-6, H1b did not down-regulate the ligand binding ability of ASGPR.The eukaryotic expression plasmid H1b/pcDNA3.1 (neo) and H2c/pcDNA3.1 (neo) were co-transfected transiently into Hela cell.Neither single H1b nor H1b and H2c could bind ASOR.In conclusion, a functional cell line of human asialoglycoprotein receptor (ASGPR) which expresses both H1a and H2c stably was established.The new split variant H1b has no effect on ASGPR binding to ASOR.ASGPRH1b alone can’t bind to ASOR, it yet can’t form functional complex with ASGPRH2c.
文摘目的:分析乙型肝炎病毒(hepantitis B virus,HBV)和丙型肝炎病毒(hepantitis C virus,HCV)感染患者血清中肝特异性抗体水平,探讨其与肝细胞损伤的关系。方法:收集2011年1月至2011年10月川北医学院附属医院HBV和HCV感染患者和健康体检者血清,检测患者丙氨酸氨基转移酶(alannine aminotransferase,ALT)和总胆红素水平;通过酶联免疫吸附法检测抗肝细胞膜特异性脂蛋白(liver specific lipoprotein,LSP)和抗去唾液酸糖蛋白受体(asialoglycoprotin receptor,ASGPR)的血清水平;以ALT>40分为肝功能异常组与正常组。使用SPSS13.0统计软件分析结果,P<0.05认为有统计学意义。结果:HBV(H)与健康对照组比较anti-LSP、anti-ASGPR具有统计学差异,HBV组患者anti-ASGPR水平与ALT具有相关性。HCV(H)与健康对照组比较anti-LSP具有统计学差异,HCV组患者anti-LSP与anti-ASGPR具有相关性。HBV患者anti-LSP、anti-ASGPR表达水平较HCV患者高。结论:HBV、HCV感染后,anti-LSP和anti-ASGPR表达高于健康对照组,ALT水平升高时抗肝特异性抗体表达明显增高,表明其可能作为监测肝细胞损伤的特异性免疫学指标。
