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时间:2020-04-30
《儿童急性淋巴细胞白血病中BAFF、APRIL及其受体的表达研究.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、·950·中国免疫学杂志2014年第3O卷doi:10.3969/j.issn.1000-484X.2014.07.019儿童急性淋巴细胞白血病中BAFF、APRIL及其受体的表达研究①吴利辉孙宝兰②③徐美玉②宋红花③顾健辉③杨治平③倪红兵③(南京医科大学附属南京儿童医院,南京210008)中图分类号R733.71文献标志码A文章编号1000484x(2014)07-0950-06[摘要]目的:探讨B淋巴细胞刺激因子(BAFF)、增殖诱导配体(APRIL)及其受体在儿童急性淋巴细胞白血病(ALL)中的表达及意义。方法:培养ALL细胞株Raji和REH细胞,采用Rea
2、lTime-PCR及ELISA、Westernblot检测BAFF、APRIL及其受体mRNA及蛋白表达;运用荧光免疫细胞化学法和流式细胞术(FCM)鉴定BAFF、APRIL蛋白在Raji和REH细胞的定位与表达;同时运用RT.PCR和ELISA方法检测不同治疗阶段(初发20例,缓解23例)ALL患儿和健康对照儿童(15例)各指标mRNA和血浆蛋白水平,并分析BAFF、APRIL血浆蛋白水平和主要临床指标之间的相关性。结果:各指标mRNA和蛋白在ALL细胞株中均有表达,BAFF和APRIL蛋白定位于细胞膜上,FCM检测其阳性细胞比例及平均荧光强度比对照组明显升高(P
3、<0.05);初发ALL儿童血浆BAFF、APRIL及其受体mRNA和蛋白水平比健康儿童显著升高(P4、生作用。[关键词]儿童急性淋巴细胞白血病;B淋巴细胞刺激因子;增殖诱导配体ExpressionandsignificanceofBAFF/APRILinchildhoodacutelymphoblasticleu-kemiawuLi—Hui,SUNBao—Lan,XUMei—Yu,SONGHong-Hua,GUJian—Hui,YANGZhi—P,NIHong—Bing.NanjingChildrensHospitalAffiliatedtoNanjingMedicalUniversity,Ⅳnng210008,China[Abstract]Objective:To5、investigatetheexpressionandsignificanceofBcellactivatingfactor(BAFF)andaproliferation—inducingligand(APRIL)inchildrenwithacutelymphoblasticleukemia(ALL).Methods:ThemRNAandproteinexpressionsinALLcellslinesRajiandREHwereexaminedbyReal—TimequantitativePCR(RT—PCR),ELISAandWesternblotmethod6、s.ImmunoeytochemistrywasusedtoobservethelocationofBAFFandAPRILinALLcelllines.Thepercentageofpositivecellsandmeanfluorescenceintensitywereevaluatedbytechniqueofflowcytometry.Meanwhile,wedetectedthemRNAandproteinlevelsofBAFF/APRILandtheirreceptorsin43childrenwithALL(20newlydiagnosedpatie7、nts,23remissionpatients)and15casesofcontrolgroupbyELISAandRT—PCRfollowinganalysistherelationshipbetweenBAFF/APRILandmainclinicalparametersofALLchildren.Results:BothmRNAandproteinexpressionsofBAFFandAPRILandtheirreceptorsexpressedinRajiandREHcells.Immunocytofluorescencestainingshowedt
4、生作用。[关键词]儿童急性淋巴细胞白血病;B淋巴细胞刺激因子;增殖诱导配体ExpressionandsignificanceofBAFF/APRILinchildhoodacutelymphoblasticleu-kemiawuLi—Hui,SUNBao—Lan,XUMei—Yu,SONGHong-Hua,GUJian—Hui,YANGZhi—P,NIHong—Bing.NanjingChildrensHospitalAffiliatedtoNanjingMedicalUniversity,Ⅳnng210008,China[Abstract]Objective:To
5、investigatetheexpressionandsignificanceofBcellactivatingfactor(BAFF)andaproliferation—inducingligand(APRIL)inchildrenwithacutelymphoblasticleukemia(ALL).Methods:ThemRNAandproteinexpressionsinALLcellslinesRajiandREHwereexaminedbyReal—TimequantitativePCR(RT—PCR),ELISAandWesternblotmethod
6、s.ImmunoeytochemistrywasusedtoobservethelocationofBAFFandAPRILinALLcelllines.Thepercentageofpositivecellsandmeanfluorescenceintensitywereevaluatedbytechniqueofflowcytometry.Meanwhile,wedetectedthemRNAandproteinlevelsofBAFF/APRILandtheirreceptorsin43childrenwithALL(20newlydiagnosedpatie
7、nts,23remissionpatients)and15casesofcontrolgroupbyELISAandRT—PCRfollowinganalysistherelationshipbetweenBAFF/APRILandmainclinicalparametersofALLchildren.Results:BothmRNAandproteinexpressionsofBAFFandAPRILandtheirreceptorsexpressedinRajiandREHcells.Immunocytofluorescencestainingshowedt
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