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1、1及联合Hsp70-B16抗原肽复合物对小鼠黑素瘤肺转移的抑制作用!至!!星鲞墓!!期inJDermatol,November2o07.v01.40,No.11可溶性PD一1及联合Hsp70一B16抗原肽复合物对小鼠黑素瘤肺转移的抑制作用邱惠张桂梅张慧耿辉冯作化【摘要】目的探讨黑素瘤肺转移小鼠模型体内表达可溶性PD一1(sPD一1)对B7一H1/PD一1信号传导的阻断作用,及联合Hsp70一B16抗原肽对小鼠黑素瘤肺转移的抑制作用.方法建立小鼠黑素瘤肺转移模型,应用免疫组织化学染色和流式细胞术检测肺转移灶PD一1及B7一H1的表达情况,从小鼠尾静脉注射
2、sPD一1表达质粒,并联合应用Hsp70一B16抗原肽皮下免疫小鼠,于接种瘤细胞后第17天解剖小鼠,观察黑素瘤肺转移的情况,检测各组小鼠肺转移灶局部淋巴细胞浸润及部分免疫学指标.结果小鼠黑素瘤肺转移模型成功建立,肺转移灶局部有大量的PD一1阳性细胞,转移瘤B16细胞表面表达较多B7一H1分子,静脉注射sPD一1表达质粒联合抗原肽免疫治疗组小鼠肺转移明显受抑制,抑制率达95%,显着高于对照组,其对应组小鼠肺部CD8+T淋巴细胞的数量,脾淋巴细胞的细胞毒活性及血清中的IL一2和IFN一的浓度均明显高于对照组(P<0.01).结论小鼠体内转染表达sPD
3、一1可阻断B7一H1/PD一1信号传导,抑制小鼠黑素瘤肺转移,联合应用Hsp70一B16抗原肽具有明显的协同作用.【关键词】黑色素瘤,实验性;基因疗法;HSP70热休克蛋白质类;sPD一1InhibitoryeffectonpulmonarymetastasisofmelanomainmicebysolublePD.1anditscombina-tionwithHsp70-B16antigenpeptidesQrUHuzNGGui-脚ezH4^rGHui,GENGHuFENGZuo-hua."DepartmentofRadiotherapyandChe
4、motherapy,ZhongnanHospital,WuhanUniversity,Wuhan430071,China【AbstractlObjectiveToinvestigatetheblockadeeffectsofsolublePD.1(sPD.1)expressedinvoonB7.Hl/PD.1signaltransduction.andinhibitoryeffectinpulmonarymetastasisofmelanomawithcombi.nationofHsp70.B16antigenpeptidesinmice.Method
5、sThepulmonarymetastasismodelofmelanomawasestablishedinmice.Immunohistochemicalstainingandflowcytometrywereutilizedtodetecttheexpres.sionofPD.1andB7.HlrespectivelyinpulmonarymetastasislociFourdaysaftertheinoculationoftumorcells.fortymurinemodelsofpulmonarymetastasiswererandomlydi
6、videdtobeimmunizedwithnormalsodium(groupA),emptyvectorpcDNA3.1(groupB).PD1Aplasmid(groupC)respectivelyviatailveininjection.subcutaneousinjectionofHsp70-Bl6antigenpeptides(groupD)orwiththecombinationofintra-venousPD1AplasmidandsubcutaneousHsp70-B16antigenpeptides(groupE).Thelocal
7、infiltrationwithlymphocytesinpulmonarymetastasislociwasobservedandaseriesofimmunologicalparameterswereassessedl7daysaftertheinoculationoftumorcells.ResultsThemelanomapulmonarymetastasismodelwassuccessfullyestablished.TherewerealotofPD.1positivecellsintheseloci.andB7.H1moleculewa
8、smassivelyexpressedonthesurfaceofBl6cellsinmeta
