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1、1α真核表达质粒的构建及在HSV-Ⅱ核酸疫苗中的应用286?论着?ISSN1007—8738细胞与分子免疫学杂志(C—hinJCellMolImm—uno1)2006,22(3文章编号:1007—8738(2006)03—0286—04小鼠MIP.1ot真核表达质粒的构建及在HSV-II核酸疫苗中的应用邵文爱',李晓眠(.汕头大学医学院微生物与免疫学教研室,广东汕头515031;天津医科大学微生物学教研室,天津300070)Constructionofrecombinanteukaryoticexpressionplasmidofmurinechemo-kineMIP.1etandcoimmu
2、nizationwithHSV.IIDNAvaccineSHAOWen.ai',LIXiao.mianDepartmentofMicrobiologyandImmunoh)gy,ShantouUniversityMedicalCollege,Shantou51503I;DepartmentofMicrobiology,TianjinMedicalUniversity,Tianjin300070,China[Abstract]AIM:Toconstructtherecombinanteukary—oticexpressionplasmidofmurinemacrophageinflammator
3、yprotein-1a(MIP-1a)andinvestigatetheeffectofMIP-1aasanadjuvantonimmuneresponseinducedbyherpessimplexvirustypeIIglycoproteinD(HSV—IIgD)DNAvaccine.METHODS:UsingtotalRNAfrOmRAW2~.7cellsstimula-tedwithLPS.thewholecodesequenceofmurineMlP.1awasamplifiedbyreversetranscriptionpolymerasechainre-action(f=rr.P
4、CR)andinsertedintopcDNA3atHindIII/XbaIrestrictionsites.TherecombinanteukaryoticexpressionplasmidPmwastransientlyexpressedinC0S-7cellsanditsspecificitywasdemonstratedbyRT-?PCRandboydenchem?-otaxischamber.BALB/cmicewereimmunizedwithgDDNAvaccineand/orMIP-la.andtheeffectofMIP一1aongDDNAvaccinewasevaluate
5、dbydetectinganti-?HSV?-IIan?-tibody,antigen-specificlymphoproliferativeresponses,andexaminingsurvivalratesaftermicewerechallengedintrav—aginallywithHSV—II.RESULTS:Therecombinanteukaryot—icexpressionplasmidofmurineMIP-1awasconstructed.anditwasrevealedthatimmuneresponsesofHSV—IIgDDNAvaccinewereenhance
6、dbycoimmunizationwithMIP一1a.CONCLUSION:ThemurineMlP.1acanbeusedasanadjuvantofHSV-IIgDDNAvaccine.[Keywords]murinemacrophageinflammatoryprotein—la(MIP-la);herpessimplexvirustypeII(HSV—II);DNAvaccine;molecularadjuvant收稿日期:2005—04—11;修回日期:2005—10—08作者简介:邵丈爱(1968一),女,山西平遥人,讲师,博士后Tel:(0754)8900840;Email:s
7、waO01@126.conl[摘要]目的:构建小鼠巨噬细胞炎性蛋白一1n(MIP一1n)的真核表达质粒,观察其作为分子佐剂对单纯疱疹病毒II型(HSV—II)DNA疫苗免疫效果的影响.方法:以LPS刺激RAW2~.7细胞提取总RNA.采用RT—PCR扩增出MIP一1ot基因的全部编码序列,利用克隆载体pUCm—T,将其亚克降人peDNA3中,构建出小鼠MIP一1ot的真核表达质粒Pm;将其转染CO
