《肺癌相關(guān)抗原RNA轉(zhuǎn)染樹(shù)突狀細(xì)胞誘導(dǎo)細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞殺傷活力的體外研究》.pdf

《《肺癌相關(guān)抗原RNA轉(zhuǎn)染樹(shù)突狀細(xì)胞誘導(dǎo)細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞殺傷活力的體外研究》.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。

1、醫(yī)技雜志2015年7月第22卷箜2J0塑￡￡墮￡iges!Y21!：22N011·695·肺癌相關(guān)抗原RNA轉(zhuǎn)染樹(shù)突狀細(xì)胞誘導(dǎo)細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞殺傷活力的體外研究湖南省懷化市第二人民醫(yī)院(41800O)劉伯軒戴北鴻向啟德廖東承高細(xì)強(qiáng)吳玉蘭周頻【摘要】目的研究NCI．H1975人肺腺癌細(xì)胞株相關(guān)抗原RNA轉(zhuǎn)染的人樹(shù)突狀細(xì)胞(DCs)在體外誘導(dǎo)細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞(CIK)對(duì)人肺腺癌細(xì)胞的殺傷活力。方法使用血細(xì)胞單采機(jī)采集外周血單個(gè)核細(xì)胞(PBMCs)并利用密度梯度離心法純化獲得的單個(gè)核細(xì)胞，細(xì)胞培養(yǎng)瓶貼壁獲得單核細(xì)胞，添加重

2、組人粒細(xì)胞、巨噬細(xì)胞集落刺激因子(rhGM—CSF)和重組人白細(xì)胞介素．4(rhIL．4)誘導(dǎo)為未成熟DCs，提取NCI．H1975細(xì)胞總RNA轉(zhuǎn)染DCs后誘導(dǎo)為成熟DCs。轉(zhuǎn)染誘導(dǎo)后成熟DCs與PBMCs培養(yǎng)的CIK細(xì)胞混合培養(yǎng)后誘導(dǎo)，再次與靶細(xì)胞混合培養(yǎng)以觀察殺傷活力。實(shí)驗(yàn)分轉(zhuǎn)染肺癌相關(guān)抗原RNA的DC—CIK組、未轉(zhuǎn)染DC—CIK組及轉(zhuǎn)染空脂質(zhì)體DC—CIK組，分別采用流式細(xì)胞分析法檢測(cè)DC及CIK細(xì)胞表面抗原表達(dá)、采用CSFE腆化丙啶(PI)熒光雙標(biāo)法檢測(cè)3組殺傷腫瘤活性的差異。結(jié)果轉(zhuǎn)染相關(guān)抗原RNA組、未轉(zhuǎn)染組及轉(zhuǎn)染空

3、脂質(zhì)體組的DC及CIK細(xì)胞表面抗原表達(dá)明顯上調(diào)，轉(zhuǎn)染相關(guān)抗原RNA組與其他2組對(duì)肺癌原代細(xì)胞殺傷活力明顯增強(qiáng)。結(jié)論肺癌相關(guān)抗原RNA轉(zhuǎn)染DC誘導(dǎo)CIK細(xì)胞對(duì)靶細(xì)胞的殺傷活力明顯增強(qiáng)，可為臨床應(yīng)用個(gè)體化DC．CIK細(xì)胞過(guò)繼免疫治療提供新的方法。【關(guān)鍵詞】樹(shù)突細(xì)胞；細(xì)胞因子誘導(dǎo)殺傷細(xì)胞；免疫療法，過(guò)繼；腺癌，細(xì)支氣管肺泡StudyoflungcancerassociatedantigenRNA-transfectedDC-inducedcellkiltingactivityofCIK／nvitroLiuBoxuan，DaiBeiho

4、ng，XiangQide，LiaoDongcheng，GaoXiqiang，WuYulan，Zhoun．PeoplesHospitalofHuaihuaC，HuI／M,／t418000，China【Abstract】objectiveToinvestigatetheNCI．H1975humanlungadenocarcinomacel1lineassociatedantigenRNA—transfectedhumandendriticcells(DCs)irtvitrocytokine—inducedkiller(CIK)act

5、ivityinducedkillercellstocancercells．MethodsUsingbloodcellapheresismachinecollectingperipheralbloodmononuclearcells(PBMCs)andusingmononuclearcellsbydensitygradientcentrifugationpurified，celcultureflasksadherentmononuclearcells，addingrecombinanthumangranulocyte—macrop

6、hagecolony—stimulatingfactor(rhGM—CSF)andrecombinanthumaninterleukin一4(rhlL一4)toinduceimmatureDCs，extractionoftotalRNANCI．H1975cellsweretransfectedDCsinducedtomatureDCs．Atiertransfectioni~ducematureDCsandPBMCsculturedCIKcellsinducedmixedculture，mixedculturewithtarget

7、cellsagaintoobservethedestructionofvitality．TheexperimentalpointsassociatedantigenRNA．transfectedDC．CIKgroupoflungcancer，non—transfectedDC—CIKgroupandtransfectedwithemptyliposomesDC—CIKgroupweredetectedbyflowcytometryanalysisDCandCIKcellsurfaceantigenexpression，using

8、CSFE／PIdoublestainingwasusedtodetectdifferencesinanti—tumoractivityofthethreegroups．ResultsRNA．a(chǎn)ssociatedantigentransfectedgroup，DC