简介:Toexploretheroleofnuclearfactor-κB(NF-κB)inthesignalpathwayofproteinkinaseC(PKC)regulatingtheproliferationandapoptosisofTlymphocytesinasthma.Tlymphocyteswereisolatedfromtheasthmaticmodelofguineapigsandtheasthmaticpatients.EithertheTcellsstimulatedwithPMAaloneorthosestimulatedwithPMAtogetherwithpyrrolidinedithiocarbamate(PDTC)wereincubatedfor1and24h.TheproliferationofandthepresenceofNF-κBinthecellsincubatedfor1hwereobservedbyMTTandimmunohistochemicalstaining,respectivelyAndthecellsincubatedfor24hwereobservedfortheapoptosisbyTUNEL.Alltheassayswereparalleledwithcontrols,andallthedatawereanalyzedstatisticallywiththesoftwareSAS.ThepercentageofcellsofnuclearpositivestainingofNF-scBandtheproliferationofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithPMAweresignificantlyhigherthanthoseofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithoutPMArespectively(P<0.01)andthoseofTlymphocytesfromnormalcontrolguineapigsandnormalcontrolpersonsstimulatedwithPMArespectively(P<0.01),andweresignificantlyreducedbyPDTC(P<0.01).TheapoptosisindexofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithPMAweresignificantlylowerthanthoseofTlymphocytesfromasthmaticguineapigsandasthmaticpatientsstimulatedwithoutPMArespectively(P<0.01)andthoseofTlymphocytesfromnormalcontrolguineapigsandnormalcontrolpersonsstimulatedwithPMArespectively(P<0.01),andweresignificantlyinducedbyPDTC(P<0.01).ThereweregoodpositivecorrelationbetweenthepercentageofcellsofnuclearstainingofNF-κBofTlymphocytesandtheproliferationofTlymphocytes(r=0.51-0.72,P<0.001),andalsogoodnegativecorrelationbetweenthepercentageofcellsofnuclearstainingofNF-scBandtheapoptosisindexofTlymphocytes(r=-0.55-0.71,P
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:摘要:目的:探讨还原型谷胱甘肽治疗病毒性肝炎的临床疗效评价。方法:选取我院2019年2月—2020年2月期间收治的72例病毒性肝炎患者作为研究对象,按照不同的治疗方式为研究组36例和对照组36例,对照组(常规治疗组)采用门冬氨酸进行治疗,研究组(还原型谷胱甘肽组)采用原型谷胱甘肽进行治疗,治疗后对组对两组病毒性肝炎的肝功能指标进行对比分析。结果:对两组病毒性肝炎患者治疗后,研究组36例患者治疗有效率为91.6%,高于对照组的77.7%,在针对疾病的治疗效果方面对比,研究组高于对照组;研究组36例患者总胆红素为(59.88±10.11)µmol/L,丙氨酸氨基转移酶为(73.68±12.14)IU/L,对照组为(83.67±11.64)µmol/L,(84.68±17.11)IU/L,肝功能指标检查结果总体优于对照组,比中差异较显著(P<
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: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: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: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:AbstractToinvestigatetheroleofCD4^+helperT(Th)cellsinthememoryCTL-mediatedanti-tumorimmunity,theRAG-1geneknockoutmicewereadoptivelytransferredwithOT-1cellstogeneratethememoryCTL,theC57B1/6miceimmunizedwiththeepitopepeptideofOVAspecificThcellsandwithdifferentadjuvantswereadopfivelytransferredwiththesememory-CTLs,andthentheanimalswerechallengedwithtumorcellsEGT.ItwasfoundthatalthoughthesimpleimmunizationofmicewiththeepitopepeptideoftheOVAspecificThcellscouldgeneratemoreeffectCTL,butthiseffectwasnotsostrongenoughtoresistcompletelythechallengeswithtumorcells.Nevertheless,thememoryCTL-mediatedanti-tumorimmuneeffectrequiredthehelpsofTh1andTh2cells.Thecross-regulationbetweenThlandTh2cellsseemedtobebeneficialforthehosttogeneratemoreeffectorCTLformountinganefficientanti-tumorresponse.ItconcludedthattheinteractionbetweenThlandTh2cellsmightbemoreimportantthanthesinglesubsetofThcellsinthememoryCTL-mediatedanti-tumorimmuneresponse.Moreattentionshouldbepaidinthisregardforthefuturestudies.
简介:【摘要】目的:于急性脑梗死患者的静脉溶栓中阿替普酶的应用效果进行评估。方法:对2019年2月至2020年2月来我院就诊的60例患者进行分组(双盲法)研究,乙组和甲组,各30例。乙组以常规治疗为主,甲组在乙组基础上加阿替普酶静脉溶栓治疗,比较神经功能缺损程度、疗效、凝血功能、炎症因子。结果:实施不同治疗后,甲组神经功能缺损程度评分小于乙组(P<0.05);甲组治疗总有效率大于乙组(P<0.05);甲组活化部分凝血酶时间、凝血酶时间、凝血酶原时间都优于乙组(P<0.05);甲组肿瘤坏死因子α、白介素6都优于乙组(P<0.05)。结论:于急性脑梗死患者中,实施阿替普酶静脉溶栓可得到确切的疗效,可明显改善患者的神经功能缺损程度、炎症因子水平、凝血功能。
简介:TheaimofthepresentstudywastodeterminetheefficacyofimmunotherapywithdendriticcellstoelicitEBV-specificCTL-immunityinadvancedcasesofEBV-positivepatientswithnasopharyngealcarcinoma(NPC)andtodeterminethesafetyandtoxicityofthispreparation.NinecasesofhistologicallyconfirmedpatientswithNPCundergoingtreatmentwithradiologicaltherapywereenrolledinthisstudy.Dendriticcells,generatedinvitrofrombloodmonocytesofpatientswereculturedandmaturedwithcytokinesandtheninfectedwithrecombinantadenovirusvaccinecontainingEBV-latentmembraneprotein-2(Ad-LMP2).On9days'cultivationofcells,thematuredDCswereharvested,irradiatedwithCoandtheninjectedintradermallytopatientswithNPC.Theinjectionswereperformed3timestotally.Afterimmunization,theCTLresponseswereassayedbymeansofcytotoxicityandepitope-specificIFN-γproduction.Theresultsofthistrialshowedthatallpatientscouldtoleratethiskindoftreatmentwithoutanysideeffect,duringwhichmarkedincreaseofLMP2-specificCTL-responsescouldbedemonstratedin5patientsofthisgroup.AndthelevelofIgA/VCAantibodydecreasedin8of9patients,thusaccountingforabetterprognosisforthesepatients.Allpatientswillbefollowedupforanotheroneyear.Atleast,thepresentworkshowsthatintradermalvaccinationwithautologousDCsinfectedwithrecombinantAd-LMP2adenovirusisasafeprocedureinNPCpatients,inwhichthisprocedurecanenhancetheLMP2-specificCTLresponsesinpatients.Thesedataareencouragingtodevelopmoreeffectivevaccinestrategiesforthetreatmentofnasopharyngealcarcinoma.