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:保证前列腺电切术后冲洗液的温度不因冲洗速度慢而造成冲洗液温度下降,造成膀胱痉挛,出血等症状。方法:设计一款加热包及膀胱冲洗液组件,用于前列腺电切术后,为膀胱冲洗液持续加热,保证冲洗液温度恒定在32°C-36°C之间。选择三门峡中心医院2020年4月至2021年4月前列腺电切术后患者共,按入院先后随机分组,设为实验组和对照,设为实验组和对照组,。 两组患者留置导尿前均无泌尿系统感染征象,且肾功能基本正常,在年龄、病程、体征等方面均无统计学意义 (P >0.05),具有可比性.比较两组使用加热包及膀胱冲洗组件后患者膀胱痉挛及泌尿系感染的发生率,比较患者前列腺电切术后所用盐水的数量及冲洗时间。结果:术后的患者通过使用加热包及其组件,可以大大降低病患者在术后出现膀胱痉挛及泌尿系感染发生的概率,缩短冲洗时间,减少冲洗液用量,促进病患良好的术后预后,提高患者的满意度,值得临床广泛运用。 最终申请专利,专利号:ZL 2019 2 0483898.0
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: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.
简介:摘要:目的:本次实验将针对结核病的治疗开展结核感染 T细胞斑点实验,分析临床辅助诊断价值。方法:本次实验选取了 2019年 1月 -2019年 6月前来本院进行疾病检查及治疗的患者为对象,对疑似结核病结核感染患者开展病情诊断分析。在自愿参与实验调查的患者中,采用计算机随机数字表法,对 264疑似例患者进行病情结果讨论。对照组患者采用结核菌素检测,观察组则为 T细胞斑点实验,分析诊断效率。结果:从检测准确率上看,观察组的阳性预测值为 94.6%( 193/204),对照组为 91.8%( 167/182),其结果依然以观察组的阳性预测率更高,接近于准确数值。与此同时,观察组检测在灵敏度、特异性上分别为 90.6%( 193/213)和 78.4%( 40/51),高于对照组的 8.4%( 167/213)和 70.6%( 36/51),组间对比差异较为显著,具有统计学意义( P< 0.05)。结论:采用 T细胞斑点实验对结核患者结核感染进行临床诊断,能够更好地提升准确度,有助于患者早期开展治疗,具有较高的临床应用价值。
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:TheaimofthisstudyistoinvestigatetheeffectsofallitridinontheexpressionoftranscriptionfactorT-bet/GA-TA-3inmiceinfectedbymurinecytomegalovirus.BALB/cmicemodelsystemofmurinecytomegalovirus(MCMV)infectionwasestablished.Inwhich20modelmicewereallocatedrandomlyintoallitridintreatedgroup(n=10)andinfectedcontrolgroup(n=10).Allitridin(25mg·kg^-1·d^-1)wasusedintreatedgroupatthe24hbyintraperitonealroute(once/d×14d),andthesamevolumeofsalinesolutionwasinjectedcontrolmice.Normalcontrolmice(n=10),wereonlygivenwiththesamevolumeof0.89%sodiumchloride,withoutinfectionwithMCMV.TheexpressionlevelsoftranscriptionfactorT-bet/GATA-3mRNAweremeasuredbyRT-PCR,andtheexpressionlevelsofThelper1(Thl)cytokineIFN-γandTh2cytokineIL-10insupernatantofspleencellcultureweremeasuredbyELISA.ExperimentalresultsshowedMCMVinfectioncouldmarkedlydown-modulatetheexpressionofIFN-γandT-bet,andsignificantlyup-modulatetheexpressionofIL-10andGATA-3mRNA.AllitridincouldinduceincreasedexpressionoftranscriptionfactorT-betmRNAandThlcytokineIFN-γsignificantly(P<0.01),anddecreasedexpressionoftranscriptionfactorGATA-3mRNAandTh2cytokineIL-10markedly(P<0.01).ItisconcludedthatMCMVinfectionleadstodisequilibriumofThl/Th2cytokineexpression:thelevelofThlcytokineIFN-γdecreasessignificantlyandTh2cytokineIL-10overexpressesmarkedly.Allitridincanup-regulatetheexpressionofT-betandIFN-γ,andinhibittheexpressionofGATA-3mRNAandIL-10inMCMVinfectedmice,indicatingaTh1dominantstatewhichshouldenhancethespecificcellularimmunereactionsagainstCMVandbehelpfulforclearanceofthecytomegalovirusinhost.
简介:Twenty-oneyearsaftermalariaantigenswerefirstclonedavaccinestillappearstobealongwayoff.Therehavebeenperiodsofgreatexcitementandinmodelsystemssubunitvaccinehomologuescaninducerobustprotection.However,significantchallengesexistconcerningantigenicvariationandpolymorphism,immunologicalnon-respons-ivenesstoindividualvaccineantigens,parasite-inducedapoptosisofimmuneeffectorandmemorycellsandimmunedeviationasaresultofmaternalimmtmityandalterationsofdendriticcellfunction.