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：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：ToobservepotentialeffectoftheengineeredbonemarrowstromalcelllineQXMSC1secretingIL-6(QXMSCIL-6)onacceleratingimmnunereconstitutioninsyngeneicbonemarrowtransplantationinmice,QXMSC1wastransfectedwiththeeukaryocyticexpressionvectorpcDNAIL-6,whichcontainedhIL-6cDNAbyliposome-mediatedgenetransfectingtechnique.G418-resistanceclonewasselectedbylimitingdilution.ThehighestsecretingclonewasselectedbyELISAassayandusedinanimalexperiments.Therecipientmice(BALB/c)werelethallyirradiatedandcotransplantedsyngeneicbonemarrow(10^7/mice)andtheQXMSCIIL-6(5×10^5/mice).LymphocyteproliferationinducedbyConAandLPS,helperTlymphocyteprecursor(HTLp),cytotoxicTlymphocyteprecursor(CTLp),plaque-formingcell(PFC),delayedtypehypersensitivity(DTH)wereexamined30,60daysinposttransplantationrespectively.TheresultsshowedthatlymphocytesproliferationtoConAandLPS,HTLp,CTLpincreased,DTHandPFCwereimprovedbycograftedstromalcellsQXMSCIIL-6on30,60daysafterBMT.TheseresultsdemonstratedthatthebonemarrowstromalcelllineQXMSC1IL-6transfectedwithIL-6(QXMSC11L-6)acceleratedimmnunereconstitutioninsyngeneicbonemarrowtransplantation.

简介：ToinvestigatetheimbalancestateofhelperTlymphocytes(Th)andcytotoxicTlympho-cytes(Tc)andtherolesofTh1/Th2/Th3andTc1/Tc2cellsinrenaltransplantationrejection,theper-centagesofthesecellsinperipheralbloodof24casesofrenaltransplantationrecipientswithacutere-jectionandthedynamicchangesoftheCD4/CD8ratioweredeterminedbyflowcytometryanalysis,while30casesofhealthyindividualsweresetupascontrols.Inthesehealthycontrols,thepercentagesoftheTh1,Th2andTh3cellswere(10.45±8.15)%,(5.05±4.15)%and(3.90±3.21)%,andthoseofTc1andTc2cellswere(9.83±7.03)%and(4.51±2.17)%,respectively.However,thepercentagesofTh1andTclcellsinperipheralbloodofthestablerecipientsaftertransplantationwere(7.29±5.62)%and(7.04±5.15)%,showingdefinitereduction,whilethoseofTh2,Th3andTc2cellsshowedsignificantincrease,(6.34±5.67)%,(4.94±4.14)%and(6.86±4.42)%,respectively.Incaseofrecipientswithacuterejection,thepercentagesofTh1andTc1cellsappearedtobe(18.55±13.21)%and(15.84±11.72)%,alsoshowingsignificantincrease,butthoseofTh2,Th3andTc2cellsappearedtobereduced,(4.19±3.62)%,(3.02±2.83)%and(3.88±1.63)%,respectively.Significantdifferencescouldbedetectedamongthesethreegroups(P<0.05).TheCD4/CD8ratioincaseswithacuterejectionwashigherthanthoseofstablerecipients(2.24±0.59vs1.95±0.45),butthatofthestablerecipientsandhealthycontrols(1.98±0.31)showednoanysignificantdifference.Fromtheaboveobservation,itisevidentthatimbalancebetweenTh1,Th2andTh3withTc1andTc2cellsmayexistafterrenaltransplantationandprobably,theim-muneimbalancemaybeinducedthroughthesecretionofcytokinesINF-γbyTh1orTc1cells,Ⅱ-4byTh2andTc2cellsandTGF-βbyTh3.

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：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：ToexploretheeffectofrhlL-15onCB-CD34^+stemcellscommittingtoNKcells,CD34^+stemcellswereobtainedfromcordblood(CB)bymagnetic-assistedcellsorting(MACS)method.CD3,CD16andCD56moleculesexpressedoncellsurfaceweredetectedbyflowcytometer.MTFmethodwasusedtotestthecytotoxicityofNKcells.Theresultswerethatstemcellfactor(SCF)alonehasnoeffectonCD34^+stemcells.IL-15stimulatedCD34^+stemcellscommittoNKcells,andSCFshowedstrongsynergisticeffectwithIL-15.ItwasconcludedthatIL-15andSCFplayeddifferentrolesduringNKcelldevelopment,llr15promotedCD34^+stemcellsdifferentiatetoNKcellprecursorandSCFimprovedtheeffectsofIL-15onNKcelldifferentiation.

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：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.