资源描述:
《外周血单核细胞诱导的cd123+髓系树突状细胞的特性研究》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、外周血单核细胞诱导的CD123+髓系树突状细胞的特性研究作者:石军,王敏,万云,池田和真【摘要】 目的:探讨体外髓系培养体系中外周血单核细胞来源的CD123+髓系树突状细胞(mDC)的生物学特性。方法:分离健康人外周血单核细胞,用重组的粒/单核细胞集落刺激因子(GMCSF)和白细胞介素4(IL4)将其诱导为树突状细胞(DC)。用流式细胞术(FCM)检测DC表面共刺激分子、CD304、CD123和CD11C的表达,并用间接免疫磁珠法将其中CD123+DC加以分离纯化;激光共聚焦显微镜、扫描电镜观察CD123+DC形态;ELISA法检测CD123
2、+DC的IL12分泌量;葡聚糖吞噬试验和3HTdR渗入法分别检测CD123+DC的吞噬功能和对同种异体T细胞的刺激能力。结果:外周血单核细胞经GMCSF和IL4诱导7d后,细胞表面高度表达CD86和CD11C,中等量表达CD1a和CD123,低表达CD83,丧失CD14的表达,其中,CD123和CD11C均匀分布于DC表面。免疫磁珠纯化后的CD123+DC呈现典型的不成熟DC形态,除细胞体积较小外,其表面突起类似于CD123-DC。CD123+DC仅微量分泌IL12,其吞噬能力强于CD123-DC(P<0.05),但抗原刺激功能低于
3、后者(P<0.05)。结论:GMCSF和IL4培养体系中的CD123+DC可能是DC分化发育过程中更早期的未成熟mDC,具有独特的生物学特性。13【关键词】髓系培养体系;髓系树突状细胞;CD123 [Abstract]AIM:ToinvestigatethebiologicalcharacteristicsofCD123+DCderivedfromcordbloodmonocytesinmyeloidculturesystem.METHODS:Themonocytesisolatedfromperipheralbloodwerecult
4、uredwithGMCSFandIL4toinduceDCfor7days,andtherelatedphenotypeofDCwasanalyzedbyflowcytometery.CD123+DCwerepurifiedwithmagneticcellsortingseparationtechnique.ThemorphologicalpropertiesofCD123+DCwasobservedunderconfocallaserscanningmicroscopeandscanelectronmicroscope.Theconcent
5、rationofIL12secretedbyDCwasanalyzedbyELISA.ThefunctionofendocytosisandantigenpresentationwasevaluatedbyuptakingFITCdextranandallogeneicmixedTlymphocytereaction(MLR).RESULTS:FACSanalysisshowedthehighexpressionofCD86andCD11C,moderateexpressionofCD1aandCD123,lowlevelsofCD83onDC
6、afterinductionbyGMCSFandIL4for7days,whiletheexpressionofCD304wasratherlow.WelldistributedCD123andCD11CweredetectedonDC.PurifiedCD123+DCshowedtypicalmorphologyofimmatureDCwithsimilarshortcytoplasmicprojectionsasCD123-DC.CD123+DCsecretedlowerlevelofIL12,andexhibitedmoresigni
7、ficantendocytosisandlessantigen13presentationfunctionthanthatofCD123-DC(P<0.05).CONCLUSION:CD123+DCinducedwithGMCSFandIL4mightbemoreearlyphaseofimmaturemyeloidDCalongwiththeDCdifferentiation,withspecialbiologicalcharacteristics. [Keywords]myeloidculturedsystem;myeloidden
8、driticcell;CD123 树突状细胞(dendriticcell,DC)是功能强大的专职抗原提呈细胞,将体外培养