简介：Thecurrentconceptof“AdoptiveTCellImmunotherapyofCancer”isquitedifferentfromhowitwasoriginallyconceived．Withthedevelopmentofmoderntechnologyinmolecularbiology,cellbiology，immunologyandbiochemistryduringthelasttwentyyearsorso，adoptiveimmunotherapyhasgrownfromitsinitialformofasimple“bloodcelltransfer”intoitspresentprocesswhichinvolveshostvauccination，effectorcellactivation／polarizationandgeneticmodification．Withtheuseofimmuneadjuvantsandtheidentification／characterizationoftumor-reactiveTcellsubsets，orincombinationwithothertherapeuticstrategies，adoptivelytransferredTcellshavebecomemuchmorepotentinmediatingtumorregression．Inaddition，studiesonthetraffickingofinfusedTcells，celltransferperformedinlymphopenicmodels，aswellasthediscoveryofnoveltechniquesinimmunemonitoringforthegenerationofeffectorcellsinvitroandaftercelltransferinvivohaveprovidedusefultoolstofurtherimprovethetherapeuticefficacyofthisapproach．ThisarticlewillreviewtheserelatedaspectsofadoptiveTcellimmunotherapyofcancerwithspecificcommentsoncertaincriticalareasintheapplicationofthisapproach．Withtherapidlyevolvingadvancesinthisarea，itishopedthatthiscellularimmunologictherapyasitwasconceptualizedinthepast，canbecomemoreusefulinthetreatmentofhumancancerinthenearfuture．

简介：Tostudytheroleofnaturalkiller(NK)cellsinTcellrecruitmentinmurineliverinfectedwithvirus,micewereintravenouslyinjecteddailywithanti-NK1.1^+antibodytodepleteNKcells.Lymphocytesinthelivertissueofmiceinfectedwithtype5adenovirusdepletedintheE1andE3regionswereassessedbyfluorometricactivatedcellsorting(FACS).Ex-pressionofchemokineIP-10anditsreceptorCXCR3mRNAintheliver,hepaticlymphocytesandspleentissuewereexaminedbyreversetranscriptionpolymerasechainreaction(RT-PCR).Serumalmfineaminotransferase(ALT)wasmeasuredasanindicatorofliverinjury.Itwasfoundthatinfectionofadenovimsandanfi-Fasmonoclonalantibody(mAb)intomicecausedliverinjuryandhighexpressionofinterfemn-γinducibleprotein-10(IP-10)mRNAintheliver.Anfi-NK1.1^+mAb,whichwasintraperitoneallyinjectedintothemiceinfectedwithadenovirus,suppressesTcellrecruitmentandexpressionofIP-10mRNAinthehver.Slighterhverinjurywasalsoobserved.Afterviresinfection,expressionofCXCR3mRNAinspleenandhvertissuewasobservedatdifferenttime.TheresultssuggestedthatTcellrecruitmentwasinitiatedbyNKcelldependentchemokineIP-10,whichinducedactivatedTcellspriminginthespleentothehverofthemouse.NKcellsplayedakeyroleinTcellrecruitmentintheliverofmouseinfectedwithadenovims.

简介：Tocloneandconstructtherecombinantplasmidcontainingthemajoroutermembraneprotein(MOMP)geneofChlamydiatrachomatis(C.trachomatis)andtoexpressthefusionproteininE.coliBL21,theMOMPgenewasamphfiedbypolymerasechainreaction(PCR)fromgenomeofC.trachomatisserovarD.ThefragmentwasclonedintotheprokaryoticexpressionvectorpET-22b(+)afterdigestionwithBamHⅠandNotⅠandtransformedintoE.coliXL1-Blue.RecombinantswereselectedbyenzymedigestionandsequencingandtherecombinantplasmidwithMOMPgenewasthentransformedintoE.coliBL21withIPTGtoexpressthetargetgene.TheexpressionrecombinantproteinswerepurifiedbyNi-NTAaffinitychromatography,andidentifiedbySDS-PAGEandWesternblot.Itwasfoundthata1.2kbMOMPgenewasisolated.TheDNAsequenceofMOMPwasfoundtobejustthesameasthesequencepublishedbyGenBank.ArecombinantplasmidcontainingMOMPgenewasconstructedtoexpressthefusionproteinsinE.coli.SDS-PAGEanalysisshowedthattherelativemolecularweightoftherecombinantproteinwasabout47kDathatwasconsistentwiththetheoreticalpredictedvalue,andthespecificityoftheexpressedproteinwasconformedbyWesternblot.ItconcludedthattheMOMPgenecouldbeexpressedintheprokaryoticsystem,bywhichitprovidedthefoundationforthefuturestudiesonthebiologicalactivitiesofC.trachomatisandforthedevelopmentofvaccineagainstthispathogen.

简介：TheaimofthisstudyistofindtheexperimentalevidencethattheprecursorfrequencyofalloreactiveCTLsisproportionaltothenumberoftheT-cellepitopespecificities.ThenumberofT-cellepitopespecificitieswasmanipulatedbypulsingdifferentnumberofHLA-A2restrictedpeptide(s)ontotheT2cells,whichactedasstimulatingcellstoelicitallo-reactionbyco-culturingwithperipheralbloodlymphocytes(PBLs)ofHLA-A2negativeindividual.TenHLA-A2restrictedpeptides(allwerenormalcellcomponents)weresynthesized,andcellpeptideextractwaspreparedbyfrozenandthawed.T2cellsloadedwithdifferentnumberofpeptide(s)wereco-culturedwithPBLsofanHLA-A2negativeindividual;thelatterwerestainedwithPKH67inadvance.Thentheproliferationwasmonitoredwithflowcytometry,andtheprecursorfrequencyoftheeffectorcellswasanalyzedbytheModFitSoftware.After6dofculture,noproliferationwasobservedinthebulkcultureofPBLalone,andobviousproliferationtookplacewhenPBLsoftheHLA-A2negativewereco-culturedwithT2cellsloadedwithorwithoutloadingpeptide(s).TheprecursorfrequencyofthealloreactiveCTLswas0.052819forco-culturewithT2cellsloadedwithoutpeptide;howeveritwas0.030429forT2cellswithEBV/LMP2Aand0.030528forT2cellsloadedwithasingleautogeneicpeptide,andincreasedupto0.144942forT2cellsloadedwith10autogeneicpeptides;theprecursorfrequencywas0.203649whenco-culturedwithT2cellsloadedwithmiscellaneouspeptidesextractedfromthecytoplasmofT2cells.ThisstudyrevealsthattheprecursorfrequencyofalloreactiveCTLsisproportionaltothenumberofT-cellepitopespecificities,andindependentofthedensityoftheallogeneicHLAClassⅠmolecule.OurfindingssupportthehypothesisthatthealloreactiveTcellpopulationscomprisemiscellaneousTcellclones;eachisspecifictocorrespondingpMHC.ThenovelconstellationofpeptidespresentedbyallogeneicMHCmoleculesmakesthous

简介：ToinvestigatethechangesofimmunefunctionsandtheeffectsofAstragaiuspolysaccharide(ASP)onthecell-mediatedimmunityofthetraumaticstressmodelofmousebyamputation,50micewererandomlydividedinto5groupsforstudy,inwhichthegroupAandBservedasthenormalcontrol(byinjectonof0.5mlofsalineintra-peritoneallydaily),andasthestresscontrol(byintra-peritonealinjectonof0.5mlofnormalsalineintomiceafteramputation)respectively,tothegroupC,DandEofmice,1000mg/kg(highdose),300mg/kg(mediandose)and250mg/kg(lowdose).TheCD4^+andCD8^+Tcellsaswellastheexpressionofthec-fosproteinweredeterminedbyimmunohistochemicaltechniques,andtheexpressionsofNF-κBmRNAandIL-10mRNAwereassayedbyhybridizationinsitu.Theexperimentalresultsshowedthatincomparisonwiththenormalcontrolgroupofmice(groupA),theexpressionlevelsofNF-κBmRNA,IL-10mRNAandthec-fosproteininthetissuesofthymusandspleeninthestresscontrolsweresignificantlyelevatedandtheCD4^+TcellsandCD4/CD8ratioweredecreased.However,incomparisonwiththestresscontrolofmice(groupB),theexpressionsofNF-κBmRNAandIL-10mRNAwereinhibitedbyASP,andtheCD4^+TcellsandCD4/CD8ratiowereincreasedingroupsC,DandE,butthelevelofc-fosproteinwasdecreased.TherewasnosignificantdifferenceintheseparametersamonggroupC,DandE.Itiscon-cludedthatthefunctionsofcell-mediatedimmunityofmiceweredisturbedunderthestressconditionofthetraumaticinjuriesafteramputation.AndtheimmunefunctionscanbeeffectivelyrestoredbytheuseofAstraga/uspolysaccharide.

简介：Toinvestigatewhetherestradiol(E2)playsaroleincell-contact-dependentregulatorymechanismofTcellactivation,westudiedtheroleofE2inregulatinggenetranscriptionofCTLA-4,ICOS,B7-1,B7-2andB7hinvitro.ThespleniccellsofnormalfemaleBALB/cmicewereactivatedbyConA.ThenthecellswereculturedwithE2(100pg/mlor50ng/ml)for24hor48h,respectively.ThecellproliferationwasmeasuredbyMTTassayandtheexpressionoftheco-stimulatorymoleculesmRNAwasexaminedbyRT-PCRanalysis.WefoundthatE2(100pg/ml,physiologicallevel)stimulatedtheactivatedspleencellsproliferation;inhibitedCTLA-4,ICOS,TGF-βandIL-10genetranscription;promotedB7-1andB7-2genetranscription.E2(50ng/ml,pregnantlevel)inhibitedtheproliferationoftheactivatedspleniccells;promotedCTLA-4,B7-1,IL-10butinhibitedB7-2andTGF-βgenetranscription.Therefore,weconcludethattheeffectsofE2onTcellactivationarepartiallythroughitsregulationontheco-stimulatorymolecules.Theco-stimulatorymoleculesarecrucialcomponentsofthecell-contactdependentregulatorymechanism,andE2mayregulateTcellactivationbythismechanism.

简介：Thehousedustmites(Dermatophagoidesfarinae,Derf)arethemajorsourceofaeroallergensimplicatedintheexpressionofatopicdisorders,includingasthma,allergicrhinitisandatopicdermatitis.Inparticular,strongcircumstantialevidencesuggeststhathousedustmiteantigensareimportantprecipitatingfactorsofasthma.Manyhousedustmiteallergensareproteasesthatcanelicitairwayinflammationbystimulatingthereleaseofcytokinesfrombronchialepithelialcells.ToinvestigatewhetherDerfallergenproteasesinducedcytokineproductionfromtheepithelialcelllineBEAS-2B,BEAS-2Bcellswereculturedwith4differentconcentrationsofDerf(0.02,0.2,2,20μg/ml)for24-96h,afterwhichsupernatantswereassayedforinterleukin(IL)-6andIL-8withELISA.Reversetranscription-PCRwasalsoperformed.ThecellsheetswereintactthroughouttheobservationincontrolgroupwithoutanyexposuretoDerfantigen.IntheexperimentalgroupscellstreatedwithDerfallergenshowedchangesintheanchoragestatusofthemonolayer.Therewasasignificantincreaseinthelevelofcytokineproductioncomparedwiththeuntreatedsample.ThereleaseofIL-6andIL-8increasedinaconcentration-dependentmanner(P<0.05,respectively)withtheadditionofincreasingdosageofDerftothecellsheets.LevelsofIL-6andIL-8begantoriseat24hand48hafterallergenexposure,andtheyincreasedsignificantlyinthesupematantsat72hand96h.AtthesametimetheconcentrationdependenceofinductionofIL-6andIL-8expressionaswellasanincreaseintheexpressionofIL-6andIL-8mRNAmanifestedevidently.HDM-inducedairwayinflammationmayincludeDerf-mediatedreleaseofinflammatorymediators,andtheproteolyticactivityofanallergenmaystimulatethereleaseofproinflammatorycytokinesfromhumanbronchialepithelium.ItissuggestedthatIL-6andIL-8productionbybronchialepithelialcellsmayplayaroleinthepathogenesisofallergicasthma.

简介：ThisstudywasaimedtoobservetheexpressionofP70S6kinase(P70S6K)inoralaciniccellcarcinoma.PT0S6kinaseexpressionwasexaminedbymeansofWestern-blottestandActivityas-say.Specimenswerefrom30casesoforalaciniccellcarcinomaand15casesofnormaloraltissuewereusedascontrols.StatisticalanalysissoftwareSPSS10.0wasusedforttesttodeterminetherelationshipbetweengeneexpressionandclinicalfeatures.TheexpressionlevelofP70S6Kincreasedobviouslyinoralaciniccellcarcinomatissue(P<0.01).ActivityassaywasthesameastheWestemblottest(P<0.01).P70S6Kexpressionlevelandactivityplayedanimportantroleinthedevelopmentoforalaciniccellcarcinoma.Inconclusion,P70S6Kisamplifiedandoverexpressedinoralaciniccellcarci-nomatissue,whichsuggestsapotentialoncogenicfunction.P70S6KandotherpossibletargetsofmTORcontributesignificantlytotumordevelopmentandthatinhibitionoftheseproteinsmaybethera-peuticforcancerpatients.OverexpressionofP70S6Kmaybeinvolvedinthepathogenesisoforalacin-iccellcarcinoma.

简介：Triptolideisanatural,biologicallyactivecomponentderivedfromChineseherbTripterygiumWilfordiiHookF.(TWHF)whichiseffectiveintheclinicaltreatmentofautoimmunediseases,however,themechanismsbywhichtriptolideexertsimmunosuppressionremainfullyunderstood.Theprimaryofthisstudyistodemonstratewhethertriptolidecanaffectphenotype,cytokineproductionandallogeneicTcell-stimulatorycapacityofdendriticcells(DCs)whicharecriticalintheinductionofimmuneresponseortolerance.PhenotypicanalysisshowthattriptolidedoesnotaffecttheexpressionofMHC(Ia^b),CD80,CD86andCD40ofDCstimulatedwithornotLPS,butsignificantlyinhibitsIL12p70productionbyDCinadose-dependentmanner.Triptolide-treatedDCsexhibitareducedcapacitytostimulateproliferationofallogeneicCD4^+Tlymphocytes.Therefore,triptolide-mediatedimmunosuppressionmaydue,inpart,totheinhibitionofIL-12p70productionandimpairmentofallogeneicTcell-stimulatorycapacityofDCs.Ourresultsmayprovideapossiblemechanisticexplanationfortheeffectivenessoftriptolideinthetreatmentofautoimmunediseases.

简介：InordertoimprovethefunctionalaffinityofthehumanizedVHsingledomainantibodyagainsthumanlungcancer,thegenescodingthehomogenousdimersdihu3D3V_Handtetramerstehu3D3V_HwereconstructedbyfusingtheSVS-Cysshortpeptideandp53tetramerizationstructuraldomaingenetohu3D3V_HgeneviarecombinantPCRtechnique,respectively.Then,thedihu3D3V_Handtehu3D3V_HgeneswereclonedtotheprokaryoticexpressionvectorpET-22b(+)andexpressedinE.coliBL21(DE3).TheproteinsexpressedwerepurifiedthroughNi~(2+)-affinitychromatographiccolumn.Meanwhile,thehu3D3V_H,dihu3D3V_Handtehu3D3V_HproteinswerelabeledwithFITC,andtheirreactivitywithan-tigenandspecificitywereanalyzedbyimmunofluorescenceassay.Astotheirfunctionalaffinities,itwasanalyzedandcomparedbyflowcytometry.Theresultsindicatedthatthesetwogeneswereexpressedasmonomersandmainlyasinclusionbodies.Afterpurificationandrenaturation,therewereabout50%ofdimersand70%oftetramerremainingintheproteinsolution.Inaddition,thedihu3D3V_Handte-hu3D3V_Hproteinsstillremainedthereactivitywithantigenandspecificityofhu3D3V_Hprotein,andtheirfunctionalaffinitieswereincreasedabout60%or100%respectively,comparedwiththoseofhu3D3V_Hprotein.Itisevidentthatthefunctionalaffinityofhu3D3V_Hproteincanbegreatlyimprovedbyincreasingitsbindingvalency.

简介：Twenty-oneyearsaftermalariaantigenswerefirstclonedavaccinestillappearstobealongwayoff.Therehavebeenperiodsofgreatexcitementandinmodelsystemssubunitvaccinehomologuescaninducerobustprotection.However,significantchallengesexistconcerningantigenicvariationandpolymorphism,immunologicalnon-respons-ivenesstoindividualvaccineantigens,parasite-inducedapoptosisofimmuneeffectorandmemorycellsandimmunedeviationasaresultofmaternalimmtmityandalterationsofdendriticcellfunction.

简介：TostudythemechanismofinfectionofEpstein-Barrvirus(EBV)ingastriccarcinomacells,theAkataandP3HR-1strainsofEBVwereusedastheteststrainsofviruses,andthesignetringcelllineHSC-39ofgastriccarcinomacellswasusedasthetargetcellsofinfection.Thevirus-infectedcellcloneswereisolatedbylimiteddilutionmethod.ItwasfoundthattheEBV-encodedsmallRNA(EBER)couldbedetectedintheinfectedcells.TheAkataandP3HR-1EBVinfectedparentalcellsandmostofclonesexpressedEBNA1,butnotEBNA2.Latentmembraneprotein(LMP-1)andLMP-2,andtheQpromoter(p),butnottheCp/WpforEBNAgenetranscriptionwasactiveintheinfectedparentalcellsaswellasalltheclones.UninfectedHSC-39cellsdidnotexpressCD21,however,AkatabutnotP3HR-1EBV-infectedclonesex-pressedlowlevelofCD21mRNA.TheseresultsdemonstratethatHSC-39cellsaresusceptibletobothEBVstrainsandEBVinfectsHSC-39cellsthroughtheCD21-independentpathway.ThisstudydefinesasignetringtypeofgastriccarcinomacellslineasauniquetargetcellsforthestudyofEBVinfectionmechanism.

简介：Toinvestigatethemutationsintheupstreamregulatoryregion(URR)ofhumanpapillomavirustype16(HPV-16)fromthecervicalcancerbiopsiesinXinjiangUygurwomenanditsrelationshiptothehighincidenceofcervicalcancerinthesouthernXinjiang,thetissueDNAwasextractedfromthecervicalcancerbiopsies,andtheURRsegmentofHPV-16DNAwasamplified,sequencedandanalyzed.Thereafter,thepolymorphismofURRinHPV-16wasthenanalyzed.ItwasdemonstratedthatthepositiveratedetectedforthepresenceofURRinHPV-16was89.47%(17/19).ComparedwiththepreviouslypublishedsequenceinURRofprototypeHPV-16,somemutationsweredetectedinthesequenceofURR.Themutationsin17URRfragmentsofHPV-16couldbedividedinto11patterns(XJU-1toXJU-11)atnucleicacidlevel,inwhicheachofXJU-1andXJU-4accountedfor23.53%(4/17),andotherpatternsofmutationaccountedfor5.88%(1/17).IncomparisonwiththeURRofprototypeHPV-16,theDNAidentityofthesepatternswas98.50%-99.68%.Inthese17URRfragments,twopointmutationsoccurredatposition7192(GtoT)andposition7520(GtoA)andtheyappearedtobeconstantinXinjiangarea.ThesetwomutationswereubiquitousintheAsia-Americantypeandconferredstronginfectionactivityandcarcinogenicityofthisvirus.Inaddition,themutationsatposition7729(AtoC),position7843(AtoG)andposition7792(CtoT)couldenhanceitstranscriptionactivityconsiderably.ItisconcludedthatsomemutationsoccurinURRgeneofHPV-16inthecervicalcancerbiopsiestakenfromUygurwomeninXinjiangarea,suggestingthatcertainrelationshipexistsamongthemutationsinURRofHPV-16,thephylogenyofHPV-16andthehighincidenceofcervicalcancerinsouthernpartofXinjiangarea.

简介：Wehaveconfirmedefficientanti-tumoractivitiesoftheperipherallymphocytestransducedwithap185HER2-specificchimericT-cellreceptorgenebothinmurineandinhumaninourpreviousstudies.TofurthertestthefeasibilityofchimericT-cellreceptorinabonemarrowtransplantationmodel,wefirst,madetwomurinetumorcelllines:MT901andMCA-205,toexpresshumanp185HER2byretroviralgenetransduction.MurinebonemarrowcellswereretrovirallytransducedtoexpressthechimericT-cellreceptorandgene-modifiedbonemarrowcellsweretransplantedintolethallyirradiatedmouse.Sixmonthsposttransplantation,p185HER2-positivetumorcells:MT-901/HER2orMCA-205/HER2wassubcutaneouslyorintravenouslyinjectedtomakemousemodelssimulatingprimarybreastcancerorpulmonarymetastasis.Theinvivoanti-tumoreffectsweremonitoredbythesizeofthesubcutaneoustumororcountingthetumornodulesinthelungsafterIndiainkstaining.ThesizeofthesubcutaneoustumorwassignificantlyinhibitedandthenumberofpulmonarynodulesweresignificantlydecreasedinmouserecipientstransplantedwithchimericT-cellreceptormodifiedbonemarrowcellscomparedwiththecontrolgroup.Ourresultssuggesttheefficientinvivoanti-tumoractivitiesofchimericT-cellreceptorgenemodifiedbonemarrowcells.

简介：WehavedevelopedandtestedchimericT-cellreceptors(TCR)specificforp185HER2.Intheseexperiments,retroviralvectorsexpressingtheN297orN29ξreceptorswereconstructedinpRET6.AmphotropicviralproducercellswereestablishedintheGALV-basedPG13packagingcellline.Ficollpurifiedhumanperipheralbloodlymphocytes(PBL)werevitallytransducedusinganoptimizedprotocolincorporatingactivationwithimmobilizedanti-CD3/anti-CD28monoclonalantibodies,followedbyviralinfectioninthepresenceoffibronectinfragmentCH296.Transducedcellswereco-culturedwithhumantumorcelllinesthatoverexpress(SK-OV-3)orunderexpress(MCF7)p185HER2toassayforantigenspecificimmuneresponses.BothCD4^+andCD8^+T-cellstransducedwiththeN297orN29ξchTCRdemonstratedHER2-specificantigenresponses,asdeterminedbyreleaseofTh1likecytokines,andcellularcytotoxicityassays.OurresultssupportthefeasibilityofadoptiveimmunothempywithgeneticallymodifiedT-cellsexpressingachTCRspecificforp185HER2.

简介：TheaimofthisstudyistoinvestigatecyclinE,pl6inkdaandki67aspossiblediagnosticbiomarkersforcervicalpreneoplasiausingcervicalexfoliated-cellspecimens,andevaluatethesignificanceforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.TheexpressionofcyclinE,pl6inkdaandki67wasexaminatedin78cervicalexfoliatedepithelialspecimensdiagnosedasatypicalsquamouscellsofundeterminedsignificance(ASCUS)(12cases),cervicalintraepithelialneoplasia(CIN)oftype1(17cases),CIN2_3(38cases)andinvasivecarcinoma(11cases)usingimmunohistochemicalanalysis,andsimultaneously,theDNAstatusofhumanpapillomavims(HPY)type16/18wasdetectedbypolymerasechainreaction(PCR)usingtypespecificprimers,cyclinE,pl6inkdaandki67werealloverexpressedinCINsandinvasivecarcinoma,comparedwithlittleexpressioninASCUS(P<0.005).OverexpressionofcyclinEwasobservedinCIN1(94.1%,X^2=21.16,P<0.01),andp16inkdaandki67wereoverexpressedininvasivecarcinoma(100%and90.9%respectively).Thedegreeofpl6inkdaandki67expressioncorrelatedwellwiththatofepitheliallesions(P<0.005).HPV16/18infectionwasassessedinC1Nsandinvasivecarcinomasamples,andrevealedasignificantrelationshipwiththedegreeofcervicalepitheliallession.Theexpressionlevelofpl6inkdaandki67seemedmorecloselyassociatedwithHPVI6infectionthanthatofcyclinE(rs=1.0vsrs=0.4).Only1caseinCINIanddcasesinCIN2-3ofHPV18positivesamplesweredetected.Thereforenostatisticalsignificancewasfoundbystatisticalanalysis.OverexpressionofcyclinE,pl6inkdaandki67inCINsandinvasivecarcinomacellsdemonstratesthepotentialuseofcyclinE,pl6inkdaandki67asdiagnosticbiomarkersforHPV-relatedcervicalneoplasticlesions.Inaddition,thistechniquecanbeusedforscreeningpatientsathighriskofdevelopingcervicalcarcinoma.

简介：TheproteomicsofthedifferentialproteinexpressionsinhumangliomacelllineU251cellsinfectedwithhumancytomegalovirus(HCMV)wasinvestigatedattheproteinlevelbyusingthesurfaceenhancedlaserdesorption/ionization(SELDI)proteinchipsysteminordertodevelopamethodofstudyforthepathogenesisofHCMVinfection.Inthisstudy,theculturedU251cellswereinfectedwithHC-MVingoodconditionandthesupernatantsoflysatesandtheextracellularfluidsofthecultivatedinfect-edcellswerequantitativelydefinedfortheexpressedproteins.TheproteomicsofthedifferentialproteinexpressionincellsbeforeandafterinfectionwasanalyzedbyWCX2arraysontheproteinchipreader.Itwasdemonstratedthatthecytopathiceffectsofinfectedcellsappearedonthe5thdayafterinfection,however,thedifferentialproteinexpressionwasevidentat6hafterinfectionasrevealedbyRT-PCRandmassspectrometry.Theproteinpeakscapturedfromdifferentbatchesofsamples,fromthesamesampledetectedwithdifferentarraysorforthedifferentlimeswereallequivalent.Withthemolecularweightrangefrom2000Dato3000Da,chipcaptured82peaksfromtheintracellularfluidsand11proteinpeakfromthecellularfluidinwhichcomparedwiththecontrolgroup,theproteinpeakswithmolecularweightof13536.3Da,10046.1Daand17106.2Dawereclosetothoseofβ-amyloidpro-tein,caspase-1precursorandLPS-inducedTNF-αfactorrespectively,whichshowedbriefup-regulation4hafterinfection,andcontinuedtoraise48hlater.Theseresultsinferthattheseproteinsmaybere-latedtotheapoptosisinducedbyHCMVinfection,thussuggestingthattheapeptosisinducedbyHC-MVinfectionmayplayaroleinthepathogenesisofHCMVinfection.

简介：