干細(xì)胞在血液系統(tǒng)疾病中的研究應(yīng)用英文版

HuangHe2011.4.8ResearchandApplicationofStemCellsinHematologicDiseasesContents231HematopoieticstemcelltransplantationMSCsinhematologicdiseasetreatmentReseachofiPSCsinhematologyBlood2014[Epubaheadofprint]FirstprospectivetreatmenttrialtocomparethepotentialbeneficialeffectofT-cell-repleteHRD-HSCTswithallcontemporaneousT-cellrepleteHSCTsusingMSDsandURDs.FromMarch2008toMarch2013,305patientswereincluded.FullyMSDswereavailableMSD-HSCTs(n=90)FullyMSDswereunavailableSuitablymatchedURDswereusedasthealternativeURD-HSCTs(n=116)OnlyURDswith>2mismatchingallelelociwereavailableHRD-HSCTs(n=99)1.TreatmentProtocolsandDonorSelectionHeHuang,etal.Blood2014[Epubaheadofprint]2.ConditioningRegimenAllpatientsweregivenmyeloablativeconditioning.ForpatientsreceivingMSD-HSCTorURD-HSCT:busulfan(Bu;3.2mg/kg/dondays–7to–4),cyclophosphamide(Cy;60mg/kg/dondays–3to–2).Rabbitantithymocyteglobulin(Thymoglobulin,Genzyme)wasadministeredtopatientsreceivingURD-HSCTs(4.5–6mg/kgtotaldose).ForpatientsreceivingHRD-HSCT:cytarabine(4g/m2/dondays?10to?9),Bu(3.2mg/kg/dondays–8to–6),Cy(1.8g/m2/dondays–5to–4),Me-CCNU(250mg/m2orallyonday–3)anti-T-lymphocyteglobulin(Fresenius)(2.5mg/kg/dondays?5to?2).HeHuang,etal.Blood2014[Epubaheadofprint]Results1.Patient,DiseaseandTransplantationCharacteristicsCharacteristicsMSDHSCT(n=90)URDHSCT(n=116)HRDHSCT(n=99)PvalueMSDvsURDMSDvsHRDURDvsHRDAge(median,range),years33.5(16-56)26(10-50)25(9-55)<0.001<0.0010.278Underlyingdisease,n(%)0.5560.1260.125

AML42(46.7)55(47.4)29(29.3)

ALL32(35.6)44(37.9)50(50.5)

MPAL02(1.7)2(2.0)

CML5(5.6)2(1.7)5(5.1)

MDS7(7.8)9(7.8)10(10.1)NHL4(4.4)4(3.4)3(3.0)Donor-patientgender,n(%)<0.0010.0290.003

Male-male18(20.0)57(49.1)33(33.3)

Male-female21(23.3)31(26.7)22(22.2)

Female-male25(27.8)13(11.2)31(31.3)Female-female26(28.9)15(12.9)13(13.1)Donor-patientbloodgroup,n(%)0.5510.1480.352

Identical61(67.8)74(63.8)57(57.6)

Incompatibility29(32.2)42(36.2)42(42.4)1.Patient,DiseaseandTransplantationCharacteristicsCharacteristicsMSDHSCT(n=68)URDHSCT(n=98)HRDHSCT(n=68)PvalueMSDvsURDMSDvsHRDURDvsHRDRiskclassification,n(%)0.8130.2750.162

Standard35(38.9)47(40.5)30(31.3)

Highrisk55(61.1)69(59.5)69(68.7)TimefromdiagnosistoHSCT,mo0.3340.6860.539

Median(range)8(3-126)9(3-108)9(3-144)DiseasestatusatHSCTforacuteleukemia/lymphoma,n(%)0.570.7330.12

CR155(70.5)80(76.2)57(67.9)

CR214(17.9)13(12.4)20(23.8)

≥CR32(2.6)5(4.8)1(1.2)

Advancedstage7(9.0)7(6.7)6(7.1)MedianMNCs,×108/kg(range)10.15

(0.84-18.66)9.3

(2.37-27.25)10.47

(0.17-24.2)0.6430.7010.428MedianCD34+count,×106/kg(range)3.908

(0.715-14.1)4.7(0.86-29.64)3.36

(0.8-14.13)0.0120.13<0.001GradesII–IVandsevereaGVHDwereallsignificantlymorefrequentinpatientsundergoingURD-orHRD-HSCTcomparedwiththoseundergoingMSD-HSCT.However,theincidencesofII–IVandsevereaGVHDwerecomparableinpatientsreceivingtransplantsfromHRDstothosefromURDs.2.GVHDHeHuang,etal.Blood2014[Epubaheadofprint]PatientsundergoingMRD-HSCThadthelowestincidenceofNRM.WhereaspatientsundergoingURD-HSCTorHRD-HSCThadacomparableincidenceofNRM,evenfor71patientsreceivedafully10/10HLA-matchedURDs.3.NRMHeHuang,etal.Blood2014[Epubaheadofprint]4.OSandDFSMSDtransplantationhadasuperiorOScomparedwithHRDtransplantation,butequivalenttoURDtransplantation.However,5-yearOSrateswerecomparablebetweenURDandHRDtransplantation.Furthermore,5-yearDFSwasnotsignificantlydifferentforpatientsundergoingtransplantationusing3typesofdonors.HeHuang,etal.Blood2014[Epubaheadofprint]The5-yearincidenceofrelapsewassignificantlyaffectedbydonortypes.HRD-HSCTcarriedasuperiorGVLeffect,evenaftercontrollingforhigh-riskpatients.5.RelapseHeHuang,etal.Blood2014[Epubaheadofprint]RelapseisthemostleadingcauseofdeathCausesofdeathHeHuang,etal.Blood2014[Epubaheadofprint]RegenerativemedicineHematopoieticstem/progenitorcellstransplantation(HSCT)DifferentiationofMesenchymalstromalcellstotissuesofmesodermalandevennonmesodermaloriginManipulationofembryonicstemcellsdifferentiationTheabilitytoreprogramadultsomaticcellstoembryonic-likecells,whichinturncandifferentiatealongspecificlineagesFourimportantdevelopmentshaveinfluencedthefield:KeyhistoricalperiodsinHSCTsDiseasestreatedbyHSCTThenumberofadultHSCTinrecent20yearsThedevelopmentofHSCTTransplantactivityintheU.S.

1980-2010

Datafrom

CIBMTR2011

summaryTransplantsRelativeproportionofdonortypeforallo-HSCT

from1990-2008byEBMTsurveyBoneMarrowTransplant2011;46:174-191ActivityofHSCTinnineAsianCountries

1991-2006TranplantsTranplantsBoneMarrowTransplant2010;45:1682-1691DonorRegistriesforChinesePopulationTzuChiStemCellCenter,isthemostearlydonorregistryforChinesepopulation,andabout346,816donorsandfacilitatedover26,905donationsincluding1123donationstoChineseMainlandbytheendofOct,2011.ChineseMarrowDonorProgram(CMDP),hasgrowntoincludealmost1.42milliondonorsandwhichexpectedtobe2millionby2015,andhasfacilitatedover2,466URDdonations

bytheendofOct,2011.HongkongRedCrosshasestablishedthedonorregistrysinceSep,2005.DramaticincreaseinthenumberofURD-HSCTsinChinawasobservedafterCMDPstartedservicingthepublicin2001GrowthintotalnumberofdonorsinCMDPbyyearGrowthintotalnumberoftransplantsdonatedfromCMDPbyyearHongKongMedJ2009;15(suppl3):45-47CurrentURD-HSCTActivityinChinaTypesoftransplantation(datafromChineseHematopoieticStemCellTransplantationCommittee

from30BMTunitsduringJune2007toJune2008)CurrentURD-HSCTActivityinChinaAML,ALLandCMLarethemostcommondiseasesforURD-HSCT(datafromChineseHematopoieticStemCellTransplantationCommittee

from30BMTunitsduringJune2007toJune2008)1483casesofURD-HSCTs—Analysisof10yearstransplantoutcomesprovidedbyCMDPPatientscharacteristicsConditioningregimens1483casesofURD-HSCTs—Analysisof10yearstransplantoutcomesprovidedbyCMDPThecumulativeincidenceofII-IVaGVHDwas31.2%;III-IVaGVHD12.2%;cGVHDwas30.2%;extensivecGVHDwas12.1%;VOD2.5%;IPS5.9%;HC18.3%;TheOSat3months,1-year,2-year,and5-yearafter-HSCTwas84.7%,70.7%,65.2%;55.9%;1483casesofURD-HSCTs—Analysisof10yearstransplantoutcomesprovidedbyCMDPCoxregressionmodeltoidentifyriskfactorsforOSVariablesRelativerisk(95%CI)PAdvanceddisease1.221(1.988-1.371)0.001SeriousaGVHD4.169(2.688-6.467)<0.001Interstitialpneumonia2.509(1.671-3.766)<0.001VOD1.807(1.030-3.397)0.04Recipientage1.194(0.943-1.512)0.141Sex-mismatch0.935(0.788-1.108)0.437955donationsfromTzuChiStemCellsCenterFirstAffiliated

Hospital

ofZhejiangUniversitySchoolof

Medicine

(218cases)BeijingDaopeiHospital(66cases)GuangzhouNanfangHospital(145cases).twHSCTActivitiesNationalScientificandTechnologicalAdvancementAward(2ndPrize)for“Clinicalstudyonunrelateddonorallogeneticbonemarrowtransplantation”byStateCouncilofthePeople'sRepublicofChina.OneofthelargestandwellknownBMTcentersinChina.Oneof24overseasBMTcentersassignedbyNMDP.FullmembershipofEBMT(CIC401)ThelargestnumberofstemcelldonationsreceivedfromTaiwanTzuChiStemCellCenterinmainlandChinaThenumberofallo-HSCTfrom1998-2009*URD(48.3%)Sibling(34.9%)CordBlood(0.8%)Autologous(11.8%)Haplo(4.2%)HLA配型技術(shù)HLA是人類主要組織相容性抗原復(fù)合物（MHC）

編碼HLA的基因位于第6條染色體短臂P21區(qū)，包含400萬個堿基，超過200個的基因

HLAⅠ類基因（HLA-A、B、C）HLAⅡ類基因（HLA-DRB1、DQB1、DPB1）對異基因造血干細(xì)胞移植影響最大HLA基因具有高度多態(tài)性目前為止，已發(fā)現(xiàn)的等位基因：HLA-A125個，HLA-B260個，HLA-C225個，HLA-DRB1225個、HLA-DQB140個HLA配型技術(shù)和造血干細(xì)胞移植HLA的血清學(xué)配型（1998年前）HLA的基因?qū)W配型（1998年以后）血清學(xué)配型相合的患者仍有30%基因?qū)W配型不相合

HLA-Ⅰ類基因不合和移植物排斥有關(guān)

HLA-Ⅱ類基因不相合和GVHD有關(guān)多個HLA-Ⅰ類位點(diǎn)不相合及HLA-Ⅰ、Ⅱ位點(diǎn)都有不合的異基因造血干細(xì)胞移植患者長期生存率明顯下降GVHD概述發(fā)生率及后果HLA相合的親緣異基因造血干細(xì)胞移植30%-60%HLA相合的無關(guān)供者異基因造血干細(xì)胞移植:40%-90%與aGVHD直接或間接有關(guān)的移植相關(guān)死亡率高達(dá)50%在長期存活的病人中，仍有60%-80%發(fā)生cGVHD

分類移植后100天內(nèi)發(fā)生的GVHD稱aGVHD移植后100天后發(fā)生的GVHD稱cGVHDaGVHD的預(yù)防增加HLA配型的精確程度藥物經(jīng)典方法:CsA+MTXGVHD預(yù)防的新藥:MMF,FK-506,

ATGT細(xì)胞去除術(shù)體外去T,如:CD34+細(xì)胞純化體內(nèi)去T,如:自殺基因?qū)脒x擇性去T,如去除CD8+亞群共刺激通路阻滯

aGVHD的治療甲基強(qiáng)的松龍調(diào)整CsA劑量，使其血藥濃度維持在較高水平全環(huán)境保護(hù)抗感染治療支持治療二線藥物：MMF、

ATG、抗CD3單抗、FK-506ClinicalTrialsandBasicResearchonHSCTNon-HLAgeneticvariationsandoutcomesofallo-HSCTKIRCytokinegenepolymorphismsInnateimmunitygenesPharmacogeneticpolymorphismProphylaxisandtreatmentofGVHDprophylaxis:CSA+MMF+MTXImmunologicalmechanisms:Th17,γδTreg

MesenchymalstemcellinthetreatmentofGVHDImatinibinthetreatmentofcGVHDRelapseafterallo-HSCTBiologicalmechanismsProphylaxis,interventionandtreatmentOurResearchonKIR116unrelateddonortransplantsperformedbetweenFebruary2001andMay2008wereanalyzed;Thesepatientsreceivedstandardmyeloablative(n=103)ornonmyeloablative(n=13,withATG)conditioningfollowedbytransplantationfromHLAmatched[n=83]ormismatched[n=33]donors;Allpatientsreceivedstandardcyclosporine/MMF/shortcoursemethotrexateregimenasprophylaxisforaGVHD;HeHuang,etal.BoneMarrowTransplant.2010;45:1514-1521OurResearchonKIRNKcellalloreactionsfromgraftswasassociatedwitharemarkableGVLeffectinmyeloiddiseaseinURDtransplantation.Thepresenceofdonor-activatingKIR2DS3significantlyincreasedtheincidenceofrelapse.DonorAAcanreducetheriskofrelapseafterURD-HSCT,asmoreactivatingKIRsmayimpairtheimmunereconstitutionandhampertheGVLeffect.OurConclusionsHeHuang,etal.BoneMarrowTransplant.2010;45:1514-1521OurResearchonCytokineGeneGenePolymorphismRecipient/DonorDonorTypeaGVHDOutcomeTNFα-857C/Crecipientand/ordonorURD↑aGVHDandII-IVaGVHDTNFβ+252Gallele-positiverecipientand/ordonorURD↑aGVHDandII-IVaGVHDTNFRIIcodon196T/Trecipientand/ordonorURD↑aGVHD,↓relapseIL10-819C/Crecipientand/ordonorURDandSib↑aGVHDandII-IVaGVHD,↑deathinremission-592C/Crecipientand/ordonorURDandSib↑aGVHDandII-IVaGVHD,↑deathinremissionTGFβ1-509T/Trecipientand/ordonorURD↓aGVHDandII-IVaGVHDHeHuang,etal.BoneMarrowTransplant.2011;46:400-407HeHuang,etal.BiolBloodandMarrowTransplant.2011;17:542-549

OurResearchonImmunityGeneandPharmacogeneticPolymorphismsOurstudyindicatedthegenepolymorphismsofthetargetenzymeofmycophenolatemofetil(MMF),inosinemonophosphatedehydrogenase(IMPDH),haveimpactsontheriskofaGVHD.DonorT-cellcostimulatorymoleculegenepolymorphisms,cytotoxicT-lymphocyteantigen4(CTLA-4)gene,areassociatedwiththeriskofaGVHD.HeHuang,etal.BiolBloodandMarrowTransplant2011(Epubaheadofprint);The15th(2010)CongressofAPBMT.OralPresentation(SecondPrize);37th(2011)AnnualMeeting0fEBMT.OralPresentation(O176);OurResearchonGVHDCharacteristicsUnrelatedtransplant(n=138)Siblingtransplant(n=102)P-valueAge(median,range),years24

(10~50)32(14~52)<0.001

Donor-recipientgender,n(%)

Female-male35(25.4)33(32.4)0.249

Other103(74.6)69(67.6)Reasonfortransplantation,n(%)

Nonmalignantdisease6(4.3)6(5.9)0.245

Low-riskcancer117(84.8)90(88.2)

High-riskcancer15(10.9)6(5.9)HLAmatching,n(%)

Matched96(69.6)102(100)<0.001

Mismatched(1allele/2alleles)42(30.4)0Low-dose,short-coursemycophenolatemofetil+CSA+MTXisaneffectiveGVHDprophylaxisHeHuang,etal.15thAPBMT2010oralpresentation(O2-3)OurResearchonGVHDCharacteristicsUnrelatedtransplant(n=138)Siblingtransplant(n=102)P-valueNeutrophilrecovery,media(range)13(7~22)14(9~30)0.094

Plateletrecovery,media(range)16(8~64)15(8-63)0.8AcuteGVHD

044(31.9)78(76.5)<0.001

I34(24.6)11(10.8)0.007

II45(32.6)12(11.8)0.003

III~IV15(10.9)1(1.0)0.004ChronicGVHD

093(67.4)68(66.7)0.768

Limited23(16.7)23(22.5）0.310

Extensive22(15.9)11(10.8)0.343HeHuang,15thAPBMT2010oralpresentation(O2-3)Low-dose,short-coursemycophenolatemofetil+CSA+MTXisaneffectiveGVHDprophylaxisOurResearchonGVHDMesenchymalstemcellinthetreatmentofGVHD10nationalinventionpatentsaboutmesenchymalstemcellidentificationandenrichmentSupportedbyNationalHighTechnologyResearchandDevelopmentProgramofChinaImmunologicalmechanisms:Th17,γδTregSupportedbyNationalNaturalScienceFoundationofChinaClinicaltrialsandbasicresearchofimatinibinthetreatmentofcGVHDSupportedbyNationalNaturalScienceFoundationofChinaHowtheneoplasticcellsgothroughseveralregimesofselectioninallo-HSCTsetting？Keypointsinrelapsepost-HSCTBiologicalcharacteristicsofrelapsedleukemiacells？ImmuneescapemechanismsofGVL?DenovoleukemiaRadiotherapy/ChemotherapyAblationMinimalresidualdiseaseDonorhematopoieticstemcellsEngraftmentofdonorcellDonorT、NKcellsGVLRelapsedleukemia?OurResearchonRelapseafterAllo-HSCTTwogenetichitsmechanismsandmultipleCEBPAmutationsofinleukemogenesisandrelapseSupportedbyNationalNaturalScienceFoundationofChinaOurResearchonRelapseafterAllo-HSCTMultiplemutationsinCEBPAgenePatientatdenovoAMLPatientinCRPatientbeforeallo-HSCTDonor-derivedcellsinpatientDonor-derivedcellsinpatientinCRDonor-derivedcellsinDCLAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAMultiplemutationsinCEBPAgeneDonorThesamegerm-linemutationinCEBPAAllo-HSCTHeHuang,etal.Blood.2011;117:5257-5269OurResearchonRelapseafterAllo-HSCTOurResearchonRelapseafterAllo-HSCTAclinicalresearchinourgroupFromJune2005toOctober2007,

28consecutivepatientswithPh+CMLinCP1;Imatinibwasadministeredatadoseof400mg/day

for3–6monthsbeforetransplantation.Conditioningregimen:intravenousFlu30mg/m2/day(fromday–10to–5)oralBu4mg/kg(n=4)orintravenousBu3.2mg/kg(n=24)(fromday–6to–5)ATGFresenius5mg/kg/day(fromday–4to–1)Prophylacticimatinib300–400mg/daywas

commencedonday+100inengraftedpatientstoprevent

relapseandwas

discontinued12monthsaftertransplantation.RIC-HSCTcombinedwithimatinibmesylate

forCML(CP)inourGroupHeHuangetal.Leukemia2009;23(6):1171-1174.OurResearchonRelapseafterAlloHSCTRIC-HSCTcombinedwithimatinibmesylate

forCML(CP)inourGroupNon-relapsemortality:14.3%(4/28)Relapse-relatedmortality:3.57%(1/28)Overallsurvival:82.1%(23/28)Completemolecularremission:91.3%(21/23)Overallsurvivalanddisease-freesurvivalHeHuangetal.Leukemia2009;23(6):1171-1174.OurResearchonHaploidenticalHSCTOutcomesofhaplo-HSCTcomparedwithURD-andMR-HSCTinourcenterduringApril2008toJune2010DonorTypeII-IVaGVHDTRM(100days)OS(1year)Haploidentical(n=38)47.4%15.8%68.4%Unrelated(URD)(n=63)42.9%6.3%78.8%Matchedrelated(MR)(n=35)22.9%091.3%CohortscomparisionPValuePValuePValueHaplovsURDNodifferenceNodifferenceNodifferenceHaplovsMR<0.05<0.05NodifferenceHeHuangetal.2011BMTTandemMeeting.移植后復(fù)發(fā)分子機(jī)制復(fù)發(fā)患者來源的復(fù)發(fā)血液學(xué)復(fù)發(fā)>50%髓外復(fù)發(fā)20-45%供者來源的復(fù)發(fā)供者細(xì)胞白血病(DCL)0~5%供者來源第二腫瘤13reportedcasesBiolBloodMarrowTransplant2006;12:511-517TohokuJ.Exp.Med2010;220:121-126HeHuangetal,Onkologie2010;33:195-200移植后復(fù)發(fā)分子機(jī)制（二次打擊學(xué)說）FirstreportofmultipleCEBPAmutationscontributingtodonororiginofleukemiarelapseafterallogeneichematopoieticstemcelltransplantation

HeHuang,etal.Blood.2011;117:5257-5260我們移植中心對移植后復(fù)發(fā)的研究：DCL移植后復(fù)發(fā)分子機(jī)制（二次打擊學(xué)說）MultiplemutationsinCEBPAgenePatientatdenovoAMLPatientinCRPatientbeforeallo-HSCTDonor-derivedcellsinpatientDonor-derivedcellsinpatientinCRDonor-derivedcellsinDCLAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAMultiplemutationsinCEBPAgeneDonorThesamegerm-linemutationinCEBPAAllo-HSCT

HeHuang,etal.Blood.2011;117:5257-5260移植后復(fù)發(fā)分子機(jī)制（二次打擊學(xué)說）+12monthsbonemarrowrelapseBonemarrow:＞30%leukemicblasts,FABsubtypeM4

HeHuang,etal.Blood.2011;117:5257-5260移植后復(fù)發(fā)分子機(jī)制（二次打擊學(xué)說）STR:absolutedonorphenotypekaryotypeofthepatient:45,XX,der(15;22)(q10;q10)[10]karyotypeofthedonor:45,XX,der(15;22)(q10;q10)[10]DonorOriginofLeukemiaRelapse我們中心對移植后復(fù)發(fā)的研究Donorselectionfornaturalkillercellactivatinggenesleadstoareducedrateofrelapseafterallogeneichematopoieticcelltransplantationbothforacutemyeloidleukemiaandacutelymphoblasticleukemia.HeHuang,etal.37thAnnualMeetingoftheEuropeanGroupforBloodandMarrowTransplantation.(OralPresentationNumber405)ThegenotypeofT-cellcostimulatorymoleculeisassociatedwithrelapseincidenceinpatientswithacutemyeloidleukemiaafterallogeneichematopoieticcelltransplantation.

HeHuang,etal.2010AnnualMeetingoftheAmericanSocietyof

hematology.(OralPresentationNumber684)

Donor-derivedsolidmalignanciesafterhematopoieticstemcelltransplantation

Suchcasesprovideauniquediseasemodeltostudythecontributionoftransplantedhealthyallogeneicdonorbonemarrow-derivedcellsforcarcinogenesisFig.2IntheOSCCregion(B,D),87.6%and85.6%ofcellspositiveP-CK(red)withnegativeCD45antigensandcontainedY(orange)chromosomesrespectively.YellowarrowheadsindicatedCD45positive(brown)whitebloodcellswhichwerefromdonors.Intheadjacentnormalregion(A,C),theXXchromosomepatternandtumorcellswithpositiveP-CKandnegativeCD45were89.6%and0%respectively.Ourfindings:Adonor-derivedoralsquamouscarinomaafterHSCTFig.1A:awell-differentiatedoralsquamouscarcinomacellswithcellnestsandkeratinouspearlscanbeseenclearly(HE,×100).B:Highermagnificationmicroscopicfindings(HE,×400).HeHuangetal.StemCellsDev,2012Jan20;21(2):177-80

Donor:male,recipient:femaleFig3.ArchivalPCR-STRanalysisofPBMCspre-andpost-transplantationaswellasPBMCsfromthedonor(A).PCR-STRanalysisofmicrodissectedtumorcells(B).ABConfirmationofthecarcinomaoriginHeHuangetal.StemCellsDev,2012Jan20;21(2):177-80

RelapsinghematologicmalignanciesafterhaploidenticalHSCTOursummaryandhypothesisFig.1CorrelatingfactorsonrelapsinghematologicmalignanciesfollowinghaploidenticalHSCTHeHuang,etal.BiolBloodMarrowTransplant.2011,17(8):1099-111Fig.2Schematicrepresentationoflossofmismatchedhaplotype(a)andlossofheterozygosity(b).Patientswithoutmutationsinmalignantcells(c)keepcompleteremissionbecauseofpotentGVTeffect.HeHuang,etal.BiolBloodMarrowTransplant.2011,17(8):1099-111Fig.3Summaryofcurrentclinicalinterventionsafterrelapsepost-haploidenticalHSCT.治療技術(shù)（靶向藥物）

AclinicalresearchinourgroupFromJune2005toOctober2007,

28consecutivepatientswithPh+CMLinCP1.medianageof26years(range,17–49years).medianintervalfromdiagnosistotransplantwas

8months(range,4–28months).Imatinibwasadministeredatadoseof400mg/day

for3–6monthsbeforeHSCT.HeHuang,Leukemia2009;23(6):1171-1174;IntJHematol2009;89(4):445-451治療技術(shù)（靶向藥物）

Conditioningregimen:intravenousFlu30mg/m2/day(fromday–10to–5)oralBu4mg/kg(n=4)orintravenousBu3.2mg/kg

(n=24)(fromday–6to–5)ATGFresenius5mg/kg/day(fromday–4to–1)

PreventaGVHD:CsA,mycophenolate

mofetil

and

short-termMTX.

Imatinibwasadministered400–600mg/dayafter

transplantationto

treatrelapseddiseaseorgraft

failure.HeHuang,Leukemia2009;23(6):1171-1174;IntJHematol2009;89(4):445-451治療技術(shù)（靶向藥物）Non-relapsemortality:14.3%(4/28)Relapse-relatedmortality:3.57%(1/28)Overallsurvival:82.1%(23/28)Completemolecularremission:91.3%(21/23)HeHuang,Leukemia2009;23(6):1171-1174;IntJHematol2009;89(4):445-451治療技術(shù)（單克隆抗體）aGVHD的治療—TNF-αInhibitorInfliximab，抗TNF-α單抗，可與可溶性及胞膜TNF-α結(jié)合，中和循環(huán)中TNF-α并清除分泌TNF-α的T細(xì)胞；Etanercept，由人類75kDTNF受體的細(xì)胞外配體結(jié)合部分和人類IgG1Fc段連接而成，特異性地與TNF-α結(jié)合，阻斷TNF-α的作用；治療技術(shù)（單克隆抗體）Table:CRratesat4weeksaccordingtotreatmentgroup.Blood,2008,111(4):2470-2475Etanerceptplus

methylprednisolone

asinitialtherapy

foraGVHD治療技術(shù)（單克隆抗體）Blood,2008,111(4):2470-2475Etanerceptplus

methylprednisolone

asinitialtherapy

foraGVHDTreatmentofsteroid-refractoryacutegraft-versus-hostdiseasewithBasiliximabandEtanerceptinearlyperiod

gradeIVSR-aGVHD(skininvolvedinsevenpatients,gastro-intestinalinvolvedinsixpatients,liverinvolvedinfourpatients).Anti-cytokinetherapyconsistedofintravenousbasiliximab20mgondays1,4,8,15,andetanercept25mgsubcutaneoustwiceaweekfor4weeks,followedbyonceweeklyfor4moreweeksHeHuang,etal.2010AnnualMeetingoftheAmericanSocietyofhematology.(Poster)

Treatmentofsteroid-refractoryacutegraft-versus-hostdiseasewithBasiliximabandEtanerceptinearlyperiod

Allpatients(100%)achievedcompleteremission.Fiveof7(71.4%)patientsarecurrentlyalivewithamedianfollow-upof163(80–348)daysaftertransplantation,andamedianfollow-upof121(48–321)daysafterAnti-cytokinetherapy.OnlytwopatientsdiedofpulmonaryinfectionandcardiacarrestHeHuang,etal.2010AnnualMeetingoftheAmericanSocietyofhematology.(Poster)

Treatmentofsteroid-refractoryacutegraft-versus-hostdiseasewithBasiliximabandEtanerceptinearlyperiod

prospectivestudyintreatmentofgradesIII-IVSR-aGVHDskininvolvedinsevenpatients,gastro-intestinalinvolvedinsixpatients,liverinvolvedinfourpatients).

intravenousbasiliximab20mgondays1,4,8,15,andetanercept25mgsubcutaneoustwiceaweekfor4weeks,followedbyonceweeklyfor4moreweeksHeHuang,etal.37th(2011)AnnualMeeting0fEBMT.PosterOutcomes CRrate TRMHeHuang,etal.37th(2011)AnnualMeeting0fEBMT.Poster非去髓性造血干細(xì)胞移植非去髓性預(yù)處理的提出以往移植前超大劑量放化療對預(yù)防移植物排斥是必需的移植前高強(qiáng)度放化療使造血干細(xì)胞移植僅限年齡較輕，一般情況較好的患者最近高效免疫抑制劑氟噠拉賓（fludarabine)的使用

GVT機(jī)理研究的加深許多研究者開始探索強(qiáng)度較弱非去髓性預(yù)處理方案非去髓性預(yù)處理方案的適應(yīng)癥--ASHNST病例選擇的總原則移植物抗腫瘤效應(yīng)敏感的腫瘤必須在原發(fā)病進(jìn)展前產(chǎn)生GVL效應(yīng)

NST病例選擇處于緩解期的高危AML惰性腫瘤：CML慢性期、低度惡性淋巴瘤不適合NST的病例原發(fā)病進(jìn)展迅速，如：難治性急性白血病、CML急變對GVL不敏感，如：ALL、高度惡性淋巴瘤Decitabine-conditionedRegulatoryγδTCellsProvideEnhancedProtectionFromGVHDviaImprovedFoxp3StabilityandICOSUp-regulationYongxianHu,YanjunGu,LixiaSheng,HuaruiFu,KangniWu,LifeiZhang,LizhenLiu,ShanFu,YulinXu,HeHuang*BoneMarrowTransplantationCenter,theFirstAffiliatedHospital,ZhejiangUniversitySchoolofMedicine,Hangzhou,Zhejiang,China

Results1.DecitabinecanincreasetheinductionofγδTregsCFSE+PBMCsCFSE-γδTregsandγδTcellsCFSE

Results2.Decitabine-conditionedγδTregscanenhanceimmunosuppressiononhPBMCproliferationinvitro

AγδTcells:PBMCs1:1CFSEBC:CommonγδTregsD:Decitabine-conditionedγδTregs**********

ResultsProliferationrateofPBMCsandinhibitionrateofPBMCsproliferationarestatisticallydifferentbetweencommonγδTreganddecitabine-conditionedγδTreggroups.

Results3.Enhancedimmunosuppressionofdecitabine-conditionedγδTregsispartlyviahighamountofTGF-β1andIL-10productioninthesupernatantDecitabine-conditionedγδTregsproducehigheramountofIL-10thancommonγδTregs

********NSNSNSNS

CommonγδTregsDecitabine-conditionedγδTregsCommonγδTregsDecitabine-conditionedγδTregsγδTcellsCommonγδTregsDecitabine-conditionedγδTregsγδTcellsβ-actinIL-10β-actinTGF-β1γδTreg:hPBMCs

Results4.Improvedstabilityofdecitabine-conditionedγδTregspartlycontributestohighamountofTGF-β1productionCommon

γδTregsDecitabine-conditioned

γδTregs34.5%23.1%48.5%50.3%PercentageofγδTregsculturedwithoutcytokinesfor5days

Results5.ICOSup-regulationofdecitabine-conditionedγδTregspartlycontributestohighamountofIL-10productionIsotypecontrol

CommonγδTregs

Decitabine-conditionedγδTregs

Results6.Decitabine-conditionedγδTregscanenhancetheprotectionofxenogenicGVHDinhumanizedmiceImmunohistochemicalstainingwithanti-humanCD45onday10afterhPBMCstransfusion

ResultsMicesurvivaltimesindifferentgroups

ConclusionsDecitabinecombinedwiththecytokinescanincreasetheinductionofγδTregscomparedwiththecytokinesalonefromadulthPBMCsDecitabine-conditionedγδTregsprovideenhancedprotectionfromGVHDbothinvitroandinvivoTheenhancedprotectionfromGVHDofdecitabine-conditionedγδTregsisviaimprovedFoxp3stabilityandICOSup-regulationEx

vivo-ExpandedVγ9Vδ2TCellsCan