改良型人nk细胞体外扩增技术建立及car-nk细胞治疗技术初步分析-establishment of improved in vitro expansion technology of human nk cells and preliminary analysis of car - nk cell therapy technology.docxVIP

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改良型人nk细胞体外扩增技术建立及car-nk细胞治疗技术初步分析-establishment of improved in vitro expansion technology of human nk cells and preliminary analysis of car - nk cell therapy technology.docx

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改良型人nk细胞体外扩增技术建立及car-nk细胞治疗技术初步分析-establishment of improved in vitro expansion technology of human nk cells and preliminary analysis of car - nk cell therapy technology

英文摘要改良型人NK细胞体外扩增技术建立及CAR-NK细胞治疗技术初步研究AnimprovedmethodofhumanNKcellexpansioninvitroandpreliminarystudyofCAR-NKcellimmunotherapytechnologyAbstractNaturalkiller(NK)celladoptivetransferisapromisingapproachforcancerimmunotherapy;however,itsdevelopmenthasbeenhinderedbythelackofefficientmethodstoproducelargenumbersoffunctionalNKcells.Interleukin(IL)-21isanimportantmodulatorofnaturalkiller(NK)cellfunction.ThisstudyistoestablishanimprovedapproachforhumanNKcellexvivoexpansion.Accordingly,weengineeredthecelllinepAPC-mbIL21basedonpAPCwhichwasconstructedbyPersongenpreviouslywithco-expressingIL15andCD137L,toestablishmembrane-boundinterleukin(mbIL)-21expressedonthecellsurface,andusedtheseengineeredcells(namedpAPCNK)toexpandNKcellsfromtheperipheralbloodmononuclearcells.Theperipheralbloodmononuclearcells(peripheralbloodmononuclearcell,PBMC)fromfourhealthydonorswereisolatedandstimulatedwithpAPCNKandIL2simultaneously.ProliferatedcellswerecountedandtheNKcellphenotypeweredeterminedbyflowcytometryanalysis.ThenthecytotoxicityofexpandedNKcellstoK562andRPMI-8226wasexaminedbyCFSE/7-AADassay.Afterculturingforthreeweeks,NKcellsweresignificantlyamplifiedandtheaveragemultipleswere409.4.ThepurityoftheNKcells(CD3-CD56+)cellsincreasedfromlessthan30%toabove90%,meanwhilethecytotoxicactivitytoK562wasreachedto85.2%.NKcellhasthenon-antigen-specifickillingfeatures,whichlimitsitsclinicaltreatmentsignificantly.Inrecentyears,adoptivecellimmunotherapyespeciallythechimericantigenreceptor(CAR)modifiedTcelltechnologieshavesprungup,whichshowgoodtargeting,destructionandpersistencebothinvitroandinclinicaltrials,displayinggreatapplicationpotentialanddevelopmentprospects.ThusbasedonNKcellexpansionsystem,additionalstudywaspursedtodeveloptheCAR-NKcelltherapy改良型人NK细胞体外扩增技术建立及CAR-NK细胞治疗技术初步研究英文摘要technologytoincreaseNKcellactivityandendowNKcellwiththetumorantigenspecificcytotoxicity.Inthisstudy,wechoseHER2asatargettoexplorethepossibilityofCAR-NKcelltherapytechnology,andaccumulatedapreliminaryexperimentaldatawhichprovides