ⅱ类抗原反式激活因子与hladr抗原的关系及其意义论文

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1、Ⅱ类抗原反式激活因子与HLADR抗原的关系及其意义论文【摘要】本研究探讨Ⅱ类抗原反式激活因子（CIITA）和人类白细胞抗原（HLADR）表达时相的关系和差异，及STAT1α反义寡核苷酸（STAT1αAS）对CIITA和HLADR的抑制作用。分离健康志愿者外周血T淋巴细胞，给予不同剂量干扰素γ（IFNγ）后，用RTPCR法检测CIITAmRNA，NC)heparinizedperipheralbloodofnormaldonorsusingFicollHypaquedensitygradientcentrifu

2、gationandents.Toobtainamonocyteenrichedcellpopulation,PBMNCedium(Gibco)containing100U/mlpenicillin,100U/mlstreptomycinand10%heatinactivatedfetalcalfserum(FCS).Nonadherentcellsovedbyed(37℃)culturemedium.Thenadherentcellsedium.CD3positivecellsaccountedfor70%ofthenon

3、adherentcellpopulation.Interferonγ(IFNγ,ShanghaiCloneHighTechpany,China)l)culturesatdifferentfinalconcentrationof50U/mlto1200U/mlforinductionofHLADRantigenat37℃,5%CO2.TcellsRNA,andHLADRproteinent.Sampleserasechainreaction(RTPCR)TotalRNAanufacturer.Singlestrand

4、edcDNA2μgoftotalRNAina40μlreactionmixturecontaining0.2μgrandomprimerand300Ureversetranscriptase(Promega,Madison,ediumovedandreplacedlIFNγ.Thentheexperimentediumonlycontained1000U/mlIFNγandRPMI1640,notincubatedRNAbyRTPCRat14hours,orcellsentsesean±SDoffourindependen

5、texperiments.Statisticalanalysisoftheresultsedbyoneultiplerangemethod.P0.05HⅠsite.ThePCRproductsofCIITAgeneHⅠ,andtentsentsafterdigestionarker.Lane1:bandsafterenzymolysedbyBamH1(308bpand163bp).Lane2：PCRproducts(471bp)ExpressionofCIITAmRNAinTcellsExpressionofCIITAland

6、1000U/ml.:marker.CIITAgeneproductsedependentmanner.IFNγinducedHLADRproteinulationulation(Figure3).CIITAexpressionlIFNγtreatmentatvarioustimepoints(hour).InhibitionofCIITAandHLADRexpressionbyantisenseoligonucleotidesagainstSTAT1αThereRNAbetol/L.TheexpressionofCI

7、ITAmRNAol/L(P0.01),butthereol/LASgroupand20μmol/LASgroup(P0.05).TheexpressionlevelofCIITAmRNAintheASgroupRNAintheSgroupRNAinASgroup,Sgroupandthecontrolgroup※at14hours(±SD)GroupNumberofsamplesODratio(CⅡTA/GAPDH)AS40.475±0.039*▲S40.749±0.037#Control40.750±0.062※ASgroup

8、,Sgroupat5nmol/L,*P＜0.01,paredarkedlyinhibittheexpressionofHLADRantigen.Inaddition,thechangesinHLADRexpressionilartothoseinCIITAe