人脐带间充质干细胞外泌体免疫调节功能的研究

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1、万方数据·2630·主堡医堂盘查!Q!i生!旦!!旦筮!!鲞笠丝翅盟!!!丛盟』￡!i塑：垒!理塾箜：!!!!：y!!：堕：塑!：!!人脐带间充质干细胞外泌体免疫调节功能的研究刘明汪劲松刘沐芸胡祥徐军【摘要】目的探讨人脐带间充质干细胞(hUC-MSCs)分泌的外泌体(exosomes)的免疫调节能力。方法分离培养hUC-MSCs，收集第3代hue—MSCs上清，提取外泌体，Western印迹检测其表面特异标志CD81、CD9的表达，BCA法测定浓度；不同浓度外泌体与CD3／CD28单抗刺激的正常人外周血单个核细胞(PBMC)作用72h，流式检测作用前后Thl7和Treg细胞的比例变

2、化，检测对CD4+T、CD8+T细胞增殖能力的影响，ELISA检测干扰素(IFN)一^y、白细胞介素(IL)_6、肿瘤坏死因子0【(TNF一0【)、转化生长因子B1(TGF一131)的水平变化。结果hUC—MSCs外泌体高表达CD9、CD81，可明显抑制CD4+T和CD8+T细胞的增殖，升高CD4+CD25+FoxP3+Treg细胞比例，经CD3／CD28单抗刺激后CD45+CD4+Ki67+和CD45+CD8+Ki67+细胞比例为85．3％±5．6％和72．6％土6．3％，同时升高PBMC分泌的TGF一131水平，降低IFN一1、IL-6、TNF-Ot的水平(均P<0．05)。结

3、论hUC．MSCs分泌的外泌体具有免疫调节功能，有望成为一种免疫治疗的新选择。【关键词】间充质干细胞；脐带；外泌体；免疫StudyofimmunomodulatoryfunctionofexosomesderivedfromhumannlnbmcalcordmesenchymalstemcellsLiaMiag+，WangJinsong，厶uMuyun，舰Xiang，施JurL+StateKeyLaboratoryofRespiratoryDiseases，GuangzhouInstituteofRespiratoryDiseases，theFirstAffiliatedHospi

4、talofGuangzhouMedicalUniversity，GuangzhouMedicalUniversity，Guangzhou510120．吼inaCorrespondingauthors：XuJun，Email：xufeili@口iP．J舒．con；HuXiang，ShenzhenBeikeCellEngineeringResearchInstitute，Sknzhen518057．China．Email：huxiang@beike．ec【Abstract】objectiveToinvestigatetheimmumodulationabilityofexosomess

5、ecretedfromhumanumbilicalcord—derivedmesenchymalstemcells(hUC—MSCs)．MethodshUC—MSCswereisolatedandcultured．Exosomeswereisolatedfromtheculturemediaofthethird—generationhUC—MSCs．TheexpressionofspecificsurfacemarkerCD9andCD81weredetectedbyWesternblot．andtheconcentrationofhUC．MSCsexosomes(hUC—MSCs

6、—ex)wasevaluatedbyBCAassay．CD3／CD28．stimulatedperipheralbloodmononuclearcells(PBMCs)fromhealthydonorwereCO—culturedwithdifferentconcentrationofhUC．MSCs—exfor72h．ThepercentageofThl7andTregcellsandtheproliferationofCD4+andCD8+Tcellsweredetectedbyfloweytometry．ELISAwasusedtotestthelevelofIFN一^v，I

7、L-6，TNF．仅andTGF．131．ResultshUC．MSCs—exinhibitedtheproliferationofCD4+andCD8+cellsobviously．andincreasedtheproportionofCIM+CD25+FoxP3+Tregcells，withhighexpressionofCDSlandC139．AfterCD3／CD28monoclonalantibodystimulated．thepercentageofCD45