Cad对原癌基因HER-2neu基因免疫的调节作用①.pdf

·727 · Cad对原癌基因HER-2/neu基因免疫的调节作用① 关庆东 倪 晶 王立新 乔 滨 储以微 熊思东 (复旦大学上海医学院免疫学系教育部分子医学重点实验室上海基因免疫与疫苗研究中心，上海200032) 中国图书分类号 8392.12 文献标识码 A 文章编号 1000-484X(2006)08-0727-05 [摘 要〕 目的:研究Cad对HER-2/neu基因疫苗诱导的免疫应答的调节作用。方法:分别构建含人HER-2/neu胞外 区(hECD)的重组质粒TR421-hECD和hECD与Cad的融合质粒TR421-hECD-C3d3,pcDNA3-hECD-C3d3。以质粒基因免疫小 鼠，定期收集血清，以ELISA法检测抗HER-2/neu抗体，3H-TdR掺人法检测细胞免疫应答，观察Cad对hECD免疫应答的调节 作用，并以Cad对HBV免疫应答的调节作为对照。结果:hECD-00融合质粒诱导的抗HER-2/neu抗体水平显著低于hECD 质粒诱导的抗体水平，其诱导的淋巴细胞增殖反应也显著低于hECD质粒免疫组，但Cad能增强HBV基因免疫诱导的免疫应 答。结论:Cad负调控HER-2/neu基因免疫诱导的免疫应答。 〔关键词」C3d;HER-2/neu;基因免疫;负调控 Theregulationontheimmuneresponsesagainstproto-oncogeneHER-2/neufol- lowingDNAvaccinationbyCadmolecule GUANQing-Dong,NIfing，WANGLi-Xin，QIAOBin，CHUYi-Wei,XIONGSi-Dong.Departmentof1mmunolo- gy,ShanghaiMedicalCollegeofFudanUniversity,TheCenterforGeneImmunizationandVaccineResearch， Shanghai200032，China [Abstract]Objective:ToinvestigatewhetherCadcanregulatethespecificimmuneresponsesagainstHER-2/neuinducedby directlyinjectionofnakedplasmidsDNAcontainingthreetandemrepeatsofCadandtheextracellulardomainofHER-2/neu(hECD). Methods;RecombinantplasmidsTR421-hECDencodinghECDgene,TR421-hECD-C3d3andpcDNA3-hECD-C3d3encodinghECD- C3d3fusiongenewereconstructed.FemaleBALB/cmicewereimmunizedwiththeserecombinantplasmidsthreetimesatthreeweeks interval.ThelevelsofspecificIgGinseracollectedattheindicatedtimesfromeachgroupweredetermined场ELISAandspecificcel- lularresponseswereanalyzedby3H-TdRproliferationassay.Asacontrol,theregulationofCadontheimmuneresponsesagainstby HBV， evaluated.Results:C3dcouldenhancetheimmuneresponseschallengedbyHBV.However,thespecificantibodyresponse againsthECDinthegroupsprimedwithhECD-C3d3-DNAwassignificantlowerthanthatinhECD-DNAgroups(P0.05).Andthe specificlympho-proliferativeresponseinthemiceimmunizedwithhECD-OM-DNAwasalsosignificantlowerthanthatinhECD-DNA group(P0.05).Conclusion:Theseresultsindi