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1、250中华传染病杂志2004年8月第22卷第4期ChinJInfectDis,August2004,Vol22,No4论著乙型肝炎病毒对干扰素-受体表达及其信号通路的影响陈公英陈智任苏平吕国才李敏伟周慧明姚航平摘要目的探索乙型肝炎病毒(HBV)持续存在和复制对干扰素-受体(IFN-R)、IFN-/STAT-1信号及MHC-诱导表达的影响。方法采用流式细胞术、Western-blot及半定量逆转录-聚合酶链反应(RT-PCR)等方法,检测HepG2.2.15与HepG2肝
2、母细胞瘤细胞株及人正常细胞株LO2的IFN-R表达;同时检测细胞IFN-R1对IFN-的结合能力。观测不同时间段的IFN-/p-STAT1信号活化及IFN-/MHC-诱导效应。结果HepG2.2.15细胞膜结合型及全细胞内IFN-R1表达高于其母源性细胞HepG2及正常肝细胞株LO2细胞;HepG2.2.15细胞IFN-R1mRNA量显著高于HepG2及LO2细胞(t=9.35,P<0.01);HepG2.2.15细胞全细胞IFN-R2表达低于HepG2及LO2细胞;两株肿瘤细胞存在内源性p-S
3、TAT1条带;HepG2.2.15细胞IFN-/p-STAT1、IFN-/MHC-诱导效应较母源细胞低下。拉米夫定抑制HBVDNA后,可上调HepG2.2.15细胞的IFN-/MHC-表达。结论HBV持续存在和复制可降低IFN-/STAT1及IFN-/MHC-诱导表达的敏感性;并能上调IFN-R1表达、下调IFN-R2表达。关键词受体,干扰素;肝炎病毒,乙型;信号传递;组织相容性抗原类;干扰素型TheinfluenceofHBVoninterferongammareceptor1ex
4、pressionanditssignalpathwayCHENGong-ying,CHENZhi,RENSu-ping,etal.InstituteofInfectiousDisease,FirstAffliatedHospital,CollegeofMedicalScience,ZhejiangUniversity,Hangzhou310003,ChinaAbstractObjectiveToinvestigatewhetherhepatitisBviruswillinfluenceinterf
5、erongammare-ceptorexpressionandIFN-/STAT1signalofthetargetcells.MethodsTwohumanhepatoblastomacelllines,HepG2anditsHBVwholegenometransgeniccelllinenamedHepG2.2.15,aswellasnormallivercelllineLO2,wereusedintheexperiments.Expressionofinterferongammareceptor1wasd
6、eterminedbyflowcytometryandWesternblots.TheexpressionofthemRNAofinterferongammareceptor1wasalsoassessedbysem-iquantitiveRT-PCR.Theintegrityoftheinterferongammasiganalpathwayswasdeter-minedbyWesternblottoassessphosphorylatedsignaltransducerandactivatoroftranscr
7、iptionSTAT1andbyflowcytometrytoinvestigatetheup-regulationofMHCclassI.HLAclassIexpressionwereanalyzedonlamivudine-treatedHepG2.2.15byquantitativeflowcytometry.ResultsExpressionlevelofinterferongammareceptor1inthemembranewassignificantlyhigherinHepG2.2.15celll
8、inecomparedwithHepG2celllineandLO2cellline(95.7%vs55.5%,47.2%,P<0.01).Similarly,themRNAexpressionofreceptorswasdrasticallyhigherinHepG2.2.15celllinethanothertwocelllines(0.920.12v