抗原冲击致敏树突状细胞诱导特异性ctl杀伤jurkat细胞实验研究

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1、抗原冲击致敏树突状细胞诱导特异性CTL杀伤Jurkat细胞实验研究[字体：大中小]作者：林东军，方志刚，李旭东，刘加军，陆英【摘要】　　本研究以健康人外周血单核细胞为前体细胞，体外诱导其为树突状细胞（DC），分别负载Jurkat细胞冻融抗原及WT1多肽抗原，产生特异性细胞毒性T淋巴细胞（CTL），以探讨其对白血病Jurkat细胞的杀伤作用。联合应用rhGM-CSF、rhIL-4、rhTNF-α及rhsCD40L等细胞因子，密度梯度离心法、贴壁法从外周血获取单核细胞并进行诱导扩增，培养出DC，分别用冻融抗原及WT1多肽抗原冲击致敏DC。实验分4组：冻融抗原致敏

2、DC组为实验组A，WT1多肽致敏DC组为实验组B，未致敏DC组为对照组A，单核细胞组为对照组B，观察CTL对Jurkat细胞的杀伤作用。结果表明：培养出了具有典型特征的DC，它高表达CD40、CD80、CD1a及CD86等表面标志，能体外诱导强烈的同种异基因混合淋巴细胞增殖反应。实验组显示高水平杀伤率，与对照组比较均具有统计学意义（P＜0.01），实验组A、B间比较无统计学意义。结论：Jurkat细胞冻融抗原及WT1多肽抗原冲击致敏DC能有效诱导T细胞抗白血病作用，为临床研制DC疫苗提供了实验依据。【关键词】树突状细胞；Jurkat细胞；冻融抗原；WT1多肽

3、抗原；细胞毒作用；白血病　　Antigen-loadedDendriticCellsTriggerKillingEffectsofSpecificCyto-toxicTLymphocytesonJurkatCellsInVitro　　AbstractThisstudywasaimedtoinvestigatetheeffectsoftumorantigen-loadeddendriticcells(DC)stimulatingthespecificcytotoxicTlymphocytes(CTL)onJurkatcellsinvitro.Periphera

4、lbloodmononuclearcellswereisolatedbyFicolldensitygradientcentrifugationfromnormalhumanheparinizedblood，theadherentmonocyteswereculturedwithgranulocyte-macrophagecolonystimulatingfactor(GM-CSF)，interleukin-4(IL-4)，alphatumornecrosisfactor(TNF-α)andsCD40L，DCswereco-culturedwithfrozen

5、-thawedantigenofJurkatcellsorWT1peptides，andthenTcellsweretriggeredintospecificCTL.TheresultsshowedthatmostsuspendedcellsexhibiteddistinctivemorphologicalfeaturesofDCwhichexpressedCD40(96%)，CD86(97%)，CD80(77%)，CD1a(69%)，andgainedthepowerfulcapacitytostimulateproliferationofallogenE

6、iclymphocytes.Undertheeffector∶targetratioof20∶1，CTLsderivedfromcultureswithDCandfrozen-thawedantigenofJurkatcellsshowed91.1%cytotoxicityagainstJurkatcells，CTLderivedfromcultureswithDCandWT1peptidesshowed87.5%cytotoxicityagainstJurkatcells，cytotoxicityofCTLderivedfromcultureswithun

7、loadedDCagainstJurkatcellswas42.1%andcytotoxicityofmonocyteswas22.7%.CytotoxicityofCTLderivedfromculturewithfrozen-thawedantigenorWT1peptidesloadedDCwasstrongerthanthatincontrolgroups（P＜0.01）.Itisconcludedthatthetumorantigen-pulsedDCcaninduceefficientandspecificanti-tumorimmunity，m

8、ayplayagreatroleinclinical