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HLA是人類主要組織相容性抗原復合物（MHC）

編碼HLA的基因位于第6條染色體短臂P21區，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）對異基因造血干細胞移植影響最大HLA基因具有高度多態性目前為止，已發現的等位基因：HLA-A125個，HLA-B260個，HLA-C225個，HLA-DRB1225個、HLA-DQB140個HLA配型技術和造血干細胞移植HLA的血清學配型（1998年前）HLA的基因學配型（1998年以后）血清學配型相合的患者仍有30%基因學配型不相合

HLA-Ⅰ類基因不合和移植物排斥有關

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

分類移植后100天內發生的GVHD稱aGVHD移植后100天后發生的GVHD稱cGVHDaGVHD的預防增加HLA配型的精確程度藥物經典方法:CsA+MTXGVHD預防的新藥:MMF,FK-506,

ATGT細胞去除術體外去T,如:CD34+細胞純化體內去T,如:自殺基因導入選擇性去T,如去除CD8+亞群共刺激通路阻滯

aGVHD的治療甲基強的松龍調整CsA劑量，使其血藥濃度維持在較高水平全環境保護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.移植后復發分子機制復發患者來源的復發血液學復發>50%髓外復發20-45%供者來源的復發供者細胞白血病(DCL)0~5%供者來源第二腫瘤13reportedcasesBiolBloodMarrowTransplant2006;12:511-517TohokuJ.Exp.Med2010;220:121-126HeHuangetal,Onkologie2010;33:195-200移植后復發分子機制（二次打擊學說）FirstreportofmultipleCEBPAmutationscontributingtodonororiginofleukemiarelapseafterallogeneichematopoieticstemcelltransplantation

HeHuang,etal.Blood.2011;117:5257-5260我們移植中心對移植后復發的研究：DCL移植后復發分子機制（二次打擊學說）MultiplemutationsinCEBPAgenePatientatdenovoAMLPatientinCRPatientbeforeallo-HSCTDonor-derivedcellsinpatientDonor-derivedcellsinpatientinCRDonor-derivedcellsinDCLAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAAsinglegerm-linemutationinCEBPAMultiplemutationsinCEBPAgeneDonorThesamegerm-linemutationinCEBPAAllo-HSCT

HeHuang,etal.Blood.2011;117:5257-5260移植后復發分子機制（二次打擊學說）+12monthsbonemarrowrelapseBonemarrow:＞30%leukemicblasts,FABsubtypeM4

HeHuang,etal.Blood.2011;117:5257-5260移植后復發分子機制（二次打擊學說）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發的研究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（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（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（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（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結合，中和循環中TNF-α并清除分泌TNF-α的T細胞；Etanercept，由人類75kDTNF受體的細胞外配體結合部分和人類IgG1Fc段連接而成，特異性地與TNF-α結合，阻斷TNF-α的作用；治療技術（單克隆抗體）Table:CRratesat4weeksaccordingtotreatmentgroup.Blood,2008,111(4):2470-2475Etanerceptplus

methylprednisolone

asinitialtherapy

foraGVHD治療技術（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非去髓性造血干細胞移植非去髓性預處理的提出以往移植前超大劑量放化療對預防移植物排斥是必需的移植前高強度放化療使造血干細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機理研究的加深許多研究者開始探索強度較弱非去髓性預處理方案非去髓性預處理方案的適應癥--ASHNST病例選擇的總原則移植物抗腫瘤效應敏感的腫瘤必須在原發病進展前產生GVL效應

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的病例原發病進展迅速，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