单倍体相合造血干细胞移植患儿慢性移植物抗宿主病的临床及免疫学特征研究

单倍体相合造血干细胞移植患儿慢性移植物抗宿主病的临床及免疫学特征研究摘要目的：本研究旨在探讨单倍体异基因造血干细胞移植（Haplo-HSCT）术后患儿慢性移植物抗宿主病（cGVHD）的临床及免疫学特征，分析其潜在的预测因素，并评估现有治疗方案的效果，以期为临床早期预测和优化治疗提供理论依据。方法：通过收集2023年1月至2024年12月在深圳市儿童医院行Haplo-HSCT的64例患儿的临床及实验室数据，应用流式细胞术检测淋巴细胞亚群、T细胞精细亚群及免疫比浊法检测体液免疫指标，结合统计学方法分析cGVHD+组与cGVHD-组的组间差异。通过动态监测这些指标在cGVHD发生前、发生时及治疗后的变化，结合单因素和多因素Logistic回归分析，筛选与cGVHD发生相关的危险因素并构建预测模型。结果：研究发现，cGVHD阳性组患儿在辅助性T淋巴细胞百分比及CD4⁺/CD8⁺比值上显著高于cGVHD阴性组，而B淋巴细胞百分比和NK细胞计数则显著降低。体液免疫指标显示，cGVHD阳性组补体C3水平显著升高。单因素Logistic回归分析显示，辅助性T淋巴细胞百分比降低、CD4⁺/CD8⁺比值升高及补体C3水平升高是cGVHD发生的单一危险因素。Logistic回归多因素分析进一步明确辅助性T淋巴细胞百分比和补体C3水平为独立预测因子。临床抗GVHD治疗后，患儿的T细胞比例和表型从免疫紊乱趋于免疫稳态，但B细胞和NK细胞功能恢复缓慢，补体C3水平仍维持高位。结论：cGVHD的发生与T细胞过度活化、B细胞功能抑制及补体系统激活密切相关。辅助性T淋巴细胞百分比和补体C3水平可作为cGVHD发生的潜在预测指标。现有免疫抑制治疗虽能部分纠正T细胞免疫紊乱，但对B细胞功能重建和补体调控的免疫治疗方案尚需进一步探索，需要免疫稳态综合预防及治疗方案，减少cGVHD的发生及cGVHD持续时间长，造成不良临床后果的发生率。关键词：慢性移植物抗宿主病（cGVHD），单倍体相合造血干细胞移植，淋巴细胞亚群，T细胞精细分型，免疫稳态

CLINICALANDIMMUNOLOGICALCHARACTERISTICSSTUDYOFCHILDRENWITHHAPLOIDENTICALHEMATOPOIETICSTEMCELLTRANSPLANTATIONANDCHRONICGRAFT-VERSUS-HOSTDISEASEABSTRACTObjective:Thisstudyaimstoexploretheclinicalandimmunologicalcharacteristicsofchronicgraft-versus-hostdisease(cGVHD)inchildrenafterhaploidallogeneichematopoieticstemcelltransplantation(Haplo-HSCT),analyzeitspotentialpredictivefactors,andevaluatetheeffectsofexistingtreatmentregimens,withtheexpectationofprovidingatheoreticalbasisforearlyclinicalpredictionandoptimizedtreatment.Methods:Bycollectingtheclinicalandlaboratorydataof64childrenwhounderwentHaplo-HSCTatShenzhenChildren'sHospitalfromJanuary2023toDecember2024,flowcytometrywasusedtodetectlymphocytesubsets,Tcellsubgroups,andimmunoturbidimetrywasemployedtomeasurehumoralimmuneindicators.Statisticalmethodswereappliedtoanalyzetheinter-groupdifferencesbetweenthecGVHD+groupandthecGVHD-group.Bydynamicallymonitoringthechangesoftheseindicatorsbefore,during,andaftertheoccurrenceofcGVHD,combinedwithunivariateandmultivariateLogisticregressionanalysis,theriskfactorsrelatedtotheoccurrenceofcGVHDwerescreenedandapredictionmodelwasconstructed.Results:TheresearchfoundthatthechildreninthecGVHDpositivegrouphadsignificantlyhigherpercentagesofhelperTlymphocytesandtheCD4⁺/CD8⁺ratiocomparedtothoseinthecGVHDnegativegroup,whilethepercentagesofBlymphocytesandthecountsofNKcellsweresignificantlylower.ThehumoralimmuneindicatorsshowedthatthecomplementC3levelinthecGVHDpositivegroupwassignificantlyincreased.UnivariateLogisticregressionanalysisrevealedthatthedecreaseinthepercentageofhelperTlymphocytes,theincreaseintheCD4⁺/CD8⁺ratio,andtheincreaseinthecomplementC3levelwerethesoleriskfactorsforcGVHD.MultivariateLogisticregressionanalysisfurtherclarifiedthatthepercentageofhelperTlymphocytesandthecomplementC3levelwereindependentpredictors.Afterclinicalanti-GVHDtreatment,theTcellproportionsandphenotypesofthechildrentendedtoshiftfromimmunedisordertoimmunehomeostasis,butthefunctionsofBcellsandNKcellsrecoveredslowly,andthecomplementC3levelremainedatahighlevel.Conclusions:TheoccurrenceofcGVHDiscloselyrelatedtoexcessiveactivationofTcells,inhibitionofBcellfunction,andactivationofthecomplementsystem.ThepercentageofhelperTlymphocytesandthelevelofcomplementC3canbeusedaspotentialpredictiveindicatorsfortheoccurrenceofcGVHD.AlthoughcurrentimmunosuppressivetreatmentscanpartiallycorrectTcellimmunedisorders,furtherexplorationisneededforimmunotherapyregimensthatrestoreBcellfunctionandregulatethecomplementsystem.ComprehensivepreventionandtreatmentstrategiesforimmunehomeostasisarerequiredtoreducetheoccurrenceofcGVHDandthelongdurationofcGVHD,therebyloweringtheincidenceofadverseclinicalconsequences.Keywords:Chronicgraft-versus-hostdisease(cGVHD),haploidenticalhematopoieticstemcelltransplantation(haplo-HSCT),lymphocytesubsets,fineT-cellsubpopulation,Immunehomeostasis

单倍体相合造血干细胞移植患儿慢性移植物抗宿主病的临床及免疫学特征研究1前言慢性移植物抗宿主病（cGVHD）是异基因造血干细胞移植（allo-HSCT）后常见的并发症之一，发生率可达30%～70%ADDINZOTERO_ITEMCSL_CITATION{"citationID":"TpbHtigR","properties":{"unsorted":true,"formattedCitation":"\\super[1,2]\\nosupersub{}","plainCitation":"[1,2]","noteIndex":0},"citationItems":[{"id":46,"uris":["/users/17061017/items/4TURVMJW"],"itemData":{"id":46,"type":"article-journal","abstract":"Allogeneichematopoieticstemcelltransplantation(allo-HSCT)iswidelyusedtotreatmalignanthematologicalneoplasmsandnon-malignanthematologicaldisorders.Approximately,5000allo-HSCTproceduresareperformedinChinaannually.SubstantialprogresshasbeenmadeinhaploidenticalHSCT(HID-HSCT),pre-transplantationriskstratification,anddonorselectioninallo-HSCT,especiallyaftertheestablishmentofthe“BeijingProtocol”HID-HSCTsystem.Transplantindicationsforselectedsubgroupsinlow-riskleukemiaorsevereaplasticanemia(SAA)differfromthoseintheWesternworld.TheseuniquesystemsdevelopedbyChinesedoctorsmayinspiretherefiningofglobalclinicalpractice.Wereviewedtheefficacyofallo-HSCTpracticefromavailableChinesestudiesonbehalfoftheHSCTworkgroupoftheChineseSocietyofHematology,ChineseMedicalAssociationandcomparedthesestudiestotheconsensusorguidelineoutsideChina.Wesummarizedtheconsensusonroutinepracticesofall-HSCTinChinaandfocusedontherecommendationsofindications,conditioningregimen,anddonorselection.","container-title":"JournalofHematology&Oncology","DOI":"10.1186/s13045-018-0564-x","ISSN":"1756-8722","journalAbbreviation":"JHematolOncol","note":"PMID:29495966\nPMCID:PMC5833104","page":"33","source":"PubMedCentral","title":"Theconsensusonindications,conditioningregimen,anddonorselectionofallogeneichematopoieticcelltransplantationforhematologicaldiseasesinChina—recommendationsfromtheChineseSocietyofHematology","volume":"11","author":[{"family":"Xu","given":"Lanping"},{"family":"Chen","given":"Hu"},{"family":"Chen","given":"Jing"},{"family":"Han","given":"Mingzhe"},{"family":"Huang","given":"He"},{"family":"Lai","given":"Yongrong"},{"family":"Liu","given":"Daihong"},{"family":"Liu","given":"Qifa"},{"family":"Liu","given":"Ting"},{"family":"Jiang","given":"Ming"},{"family":"Ren","given":"Hanyun"},{"family":"Song","given":"Yongping"},{"family":"Sun","given":"Zimin"},{"family":"Wang","given":"Jianmin"},{"family":"Wu","given":"Depei"},{"family":"Zhou","given":"Daobin"},{"family":"Zou","given":"Ping"},{"family":"Liu","given":"Kaiyan"},{"family":"Huang","given":"Xiaojun"}],"issued":{"date-parts":[["2018",3,2]]}}},{"id":52,"uris":["/users/17061017/items/SHSQGVWM"],"itemData":{"id":52,"type":"article-journal","abstract":"Theconsensusin2018fromTheChineseSocietyofHaematology(CSH)onthemonitoring,treatment,andpreventionofleukaemiarelapseafterallogeneichaematopoieticstemcelltransplantation(HSCT)facilitatedthestandardizationofclinicalpracticesinChinaandprogressiveintegrationwiththeworld.Tointegraterecentdevelopmentsandfurtherimprovetheconsensus,apanelofexpertsfromtheCSHrecentlyupdatedthefollowingconsensus:(1)integraterisk-adapted,measurableresidualdisease(MRD)-guidedstrategyonmodifieddonorlymphocyteinfusion(DLI)andinterferon-αintototaltherapy,whichwaspioneeredandrefinedbyChineseresearchers;(2)provideadditionalevidenceofthesuperiorityofhaploidenticalHSCT(thedominantdonorsourceinChina)tomatchedHSCTforhigh-riskpopulations,especiallyforpre-HSCTMRD-positivepatients;(3)supporttherapidprogressoftechniquesforMRDdetection,suchasnext-generationsequencing(NGS)andleukaemiastemcell-basedMRDdetection;and(4)addresstheroleofnewtargetedoptionsintransplantsettings.Inconclusion,theestablishmentofa“totaltherapy”strategyrepresentsagreatstepforward.WehopethattheconsensusupdatedbyChinesescholarswillincludethelatestcutting-edgedevelopmentsandinspireprogressinpost-HSCTrelapsemanagement.","container-title":"CancerLetters","DOI":"10.1016/j.canlet.2024.217264","ISSN":"0304-3835","journalAbbreviation":"CancerLetters","page":"217264","source":"ScienceDirect","title":"Consensusonthemonitoring,treatment,andpreventionofleukaemiarelapseafterallogeneichaematopoieticstemcelltransplantationinChina:2024update","title-short":"Consensusonthemonitoring,treatment,andpreventionofleukaemiarelapseafterallogeneichaematopoieticstemcelltransplantationinChina","volume":"605","author":[{"family":"Wang","given":"Yu"},{"family":"Chang","given":"Ying-Jun"},{"family":"Chen","given":"Jing"},{"family":"Han","given":"Mingzhe"},{"family":"Hu","given":"JianDa"},{"family":"Hu","given":"Jiong"},{"family":"Huang","given":"He"},{"family":"Lai","given":"Yongrong"},{"family":"Liu","given":"Daihong"},{"family":"Liu","given":"Qifa"},{"family":"Luo","given":"Yi"},{"family":"Jiang","given":"Er-lie"},{"family":"Jiang","given":"Ming"},{"family":"Song","given":"Yongping"},{"family":"Tang","given":"Xiao-Wen"},{"family":"Wu","given":"Depei"},{"family":"Xia","given":"Ling-Hui"},{"family":"Xu","given":"Kailin"},{"family":"Zhang","given":"Xi"},{"family":"Zhang","given":"Xiao-Hui"},{"family":"Huang","given":"Xiaojun"}],"issued":{"date-parts":[["2024",11,28]]}}}],"schema":"/citation-style-language/schema/raw/master/csl-citation.json"}[1,2]。其发病机制复杂，临床表现多样，诊断和治疗存在瓶颈，伴有异质性免疫细胞激活异常和功能耦联。单倍体造血干细胞移植（Haplo-HSCT）具有供体来源广泛、供体选择灵活等优势，近年来已经在临床治疗中得到了广泛应用，成为目前治疗血液系统恶性疾病的一种既可靠又有效的热门选择ADDINZOTERO_ITEMCSL_CITATION{"citationID":"oLr0cDeX","properties":{"formattedCitation":"\\super[3]\\nosupersub{}","plainCitation":"[3]","noteIndex":0},"citationItems":[{"id":57,"uris":["/users/17061017/items/EGGIRGMK"],"itemData":{"id":57,"type":"webpage","title":"单倍体造血干细胞移植研究进展-国际输血及血液学杂志","URL":"/CN511693201906/1175894.htm","accessed":{"date-parts":[["2025",5,9]]}}}],"schema":"/citation-style-language/schema/raw/master/csl-citation.json"}[3]。随着这一技术的广泛应用，cGVHD的发生率并未显著降低，反而成为影响移植成功率和患者长期生存质量的关键因素ADDINZOTERO_ITEMCSL_CITATION{"citationID":"7AfYeS2x","properties":{"formattedCitation":"\\super[4]\\nosupersub{}","plainCitation":"[4]","noteIndex":0},"citationItems":[{"id":59,"uris":["/users/17061017/items/EYP2AM2D"],"itemData":{"id":59,"type":"article-journal","container-title":"中华实用儿科临床杂志","DOI":"10.3760/cma.j.issn.2095-428X.2018.03.012","ISSN":"2095-428X","issue":"03","language":"ch","license":"info:中华医学会和《中华医学杂志》社有限责任公司","note":"number:03\npublisher:《中华医学杂志》社有限责任公司","page":"208-212","source":"","title":"亲缘单倍体造血干细胞移植治疗59例儿童血液病的疗效","volume":"33","author":[{"family":"费新红","given":""},{"family":"王静波","given":""},{"family":"程昊钰","given":""},{"family":"殷宇明","given":""},{"family":"张维婕","given":""},{"family":"张书芹","given":""},{"family":"王筱璨","given":""},{"family":"刘梦琦","given":""},{"family":"赵杰","given":""}],"issued":{"date-parts":[["2018",2,5]]}}}],"schema":"/citation-style-language/schema/raw/master/csl-citation.json"}[4]。有研究表明，cGVHD的发生与供体和受体之间的免疫差异、移植前的预处理方案、移植后的免疫抑制治疗等多种因素密切相关ADDINZOTERO_ITEMCSL_CITATION{"citationID":"GIrZxj8o","properties":{"unsorted":true,"formattedCitation":"\\super[5\\uc0\\u8211{}7]\\nosupersub{}","plainCitation":"[5–7]","noteIndex":0},"citationItems":[{"id":67,"uris":["/users/17061017/items/B4XT35ZA"],"itemData":{"id":67,"type":"webpage","title":"Updatesinchronicgraft-versus-hostdisease:noveltreatmentsandbestpracticesinthecurrentera|BoneMarrowTransplantation","URL":"/articles/s41409-024-02370-8","accessed":{"date-parts":[["2025",5,9]]}}},{"id":64,"uris":["/users/17061017/items/R4LXM4NJ"],"itemData":{"id":64,"type":"article-journal","abstract":"Chronicgraft-vs-hostdisease(cGVHD)istheleadingcauseoflong-termmorbidityandmortalityfollowingallogeneichematopoieticstemcelltransplantation.Itpresentsasachronicinflammatoryandscleroticautoimmune-likeconditionthatmostfrequentlyaffectstheskin,oralmucosa,liver,eyesandgastrointestinaltract.BothclinicalandanimalstudieshaveshownthatmultipleTcellsubsetsincludingTh1,Th2,Th17,TfollicularhelpercellsandregulatoryT-cellsplaysomeroleincGVHDdevelopmentandprogression;BcellsalsoplayanimportantroleinthediseaseincludingtheproductionofantibodiestoHYandnuclearantigensthatcancauseserioustissuedamage.AnarrayofcytokinesandchemokinesproducedbydifferenttypesofimmunecellsalsomediatetissueinflammationanddamageofcGVHDtargettissuessuchastheskinandoralcavity.ManyofthesesameimmuneregulatorshavebeenstudiedascandidatecGVHDbiomarkers.Recentstudiessuggestthatsomeofthesebiomarkersmaybeusefulfordeterminingdiseaseprognosisandplanninglong-termclinicalfollow-upofcGVHDpatients.","container-title":"WorldJournalofTransplantation","DOI":"10.5500/wjt.v6.i4.608","ISSN":"2220-3230","issue":"4","journalAbbreviation":"WorldJTransplant","note":"PMID:28058210\nPMCID:PMC5175218","page":"608-619","source":"PubMedCentral","title":"Biologyofchronicgraft-vs-hostdisease:Immunemechanismsandprogressinbiomarkerdiscovery","title-short":"Biologyofchronicgraft-vs-hostdisease","volume":"6","author":[{"family":"Presland","given":"RichardB"}],"issued":{"date-parts":[["2016",12,24]]}}},{"id":61,"uris":["/users/17061017/items/QPR2IGEA"],"itemData":{"id":61,"type":"article-journal","abstract":"Chronicgraft-versus-hostdisease(cGvHD)isoneofthemajorcomplicationsofallogeneicstemcelltransplantation(HSCT).cGvHDisanautoimmune-likedisorderaffectingmultipleorgansandinvolvesadermatologicalrash,tissueinflammationandfibrosis.TheincidenceofcGvHDhasbeenreportedtobeashighas30%to60%andtherearecurrentlynoreliabletoolsforpredictingtheoccurrenceofcGvHD.Thereisthereforeanimportantunmetclinicalneedforpredictivebiomarkers.ThepresentreviewsummarizesthestateoftheartforgeneticvariationasapredictivebiomarkerforcGvHD.Wediscussthreedifferentmodesofactionforgeneticvariationintransplantation:geneticassociations,geneticmatching,andpharmacogenetics.Theresultsindicatethatcurrently,therearenogeneticpolymorphismsorgenetictoolsthatcanbereliablyusedasvalidatedbiomarkersforpredictingcGvHD.Anumberofrecommendationsforfuturestudiescanbedrawn.Themajorityofstudiestodatehavebeenunder-poweredandincludedtoofewpatientsandgeneticmarkers.Likeinallcomplexmultifactorialdiseases,largecollaborativegenome-levelstudiesarenowneededtoachievereliableandunbiasedresults.Someofthecandidategenes,inparticular,CTLA4,HSPE,IL1R1,CCR6,FGFR1OP,andIL10,andsomenon-HLAvariantsintheHLAgeneregionhavebeenreplicatedtobeassociatedwithcGvHDriskinindependentstudies.Theseassociationsshouldnowbeconfirmedinlargewell-characterizedcohortswithfinemapping.SomepatientsdevelopcGvHDdespiteveryextensiveimmunosuppressionandothertreatments,indicatingthatthecurrenttherapeuticregimensmaynotalwaysbeeffectiveenough.Hence,morestudiesonpharmacogeneticsarealsorequired.Moreover,allofthesestudiesshouldbeadjustedfordiagnosticandclinicalfeaturesofcGvHD.Weconcludethatfuturestudiesshouldfocusonmoderngenome-leveltools,suchasmachinelearning,polygenicriskscoresandgenome-wideassociationstudy-transcriptionmeta-analyses,insteadoffocusingonjustsinglevariants.TheriskofcGvHDmayberelatedtothesummarylevelofimmunogeneticdifferences,orwholegenomehistocompatibilitybetweeneachdonor-recipientpair.Asthenumberofgenome-wideanalysesinHSCTisincreasing,weareapproachinganerawheretherewillbesufficientdatatoincorporatetheseapproachesinthenearfuture.","container-title":"FrontiersinImmunology","DOI":"10.3389/fimmu.2020.575492","ISSN":"1664-3224","journalAbbreviation":"FrontImmunol","note":"PMID:33193367\nPMCID:PMC7604383","page":"575492","source":"PubMedCentral","title":"ReviewofGeneticVariationasaPredictiveBiomarkerforChronicGraft-Versus-Host-DiseaseAfterAllogeneicStemCellTransplantation","volume":"11","author":[{"family":"Partanen","given":"Jukka"},{"family":"Hyvärinen","given":"Kati"},{"family":"Bickeböller","given":"Heike"},{"family":"Bogunia-Kubik","given":"Katarzyna"},{"family":"Crossland","given":"RachelE."},{"family":"Ivanova","given":"Milena"},{"family":"Perutelli","given":"Francesca"},{"family":"Dressel","given":"Ralf"}],"issued":{"date-parts":[["2020",10,19]]}}}],"schema":"/citation-style-language/schema/raw/master/csl-citation.json"}[5–7]，其发病机制涉及T细胞、B细胞、NK细胞等多种免疫细胞的异常激活与功能失调，以及细胞因子网络的紊乱。免疫重建对移植成功起着关键作用，更是影响cGVHD出现的重要因素。移植后，患者的免疫系统需要重新建立，在这一过程中，免疫细胞的增殖、分化以及功能恢复，对移植是否成功和并发症的出现起着关键作用，辅助性CD4⁺T细胞亚群是免疫系统引发特异性细胞免疫应答以防范外界病原体入侵的核心力量之一，其功能的恢复、平衡对维持免疫的稳态是至关重要的ADDINZOTERO_ITEMCSL_CITATION{"citationID":"iOGajYHo","properties":{"unsorted":true,"formattedCitation":"\\super[8,9]\\nosupersub{}","plainCitation":"[8,9]","noteIndex":0},"citationItems":[{"id":69,"uris":["/users/17061017/items/DFMEL42E"],"itemData":{"id":69,"type":"article-journal","abstract":"Immunosuppressivemediatorsintuberculosispleurisy(pleuralfluid(PF))areassociatedwiththecourseofdisease,buttheyremainpoorlydefined.Tostudythelocalimmunestatusofpatientswithtuberculosispleurisy,weexaminedtheeffectofPFonthefunctionsofTcellsandthedifferentiationofTh1cells.PFcouldinhibittheabilityofTcellstoproducecytokines.However,tumor-necrosisfactor(TNF)-αderivedfromnon-Tcellswasnotimpaired.Furtheranalysisindicatedthatcellactivationandcellcycleprogressionwerealsosuppressed.Moreover,PFcouldinhibitTh1celldifferentiation.Importantly,wefoundthatinhibitorsofindoleamine2,3-dioxygenase(IDO)andadenosineandneutralizingantibodiesagainstIL-10andtransforminggrowthfactor(TGF)-βcouldreversecytokineproduction,suggestingthatIDO,adenosine,IL-10andTransforminggrowthfactor–β1inPFmighttakepartinimpairingT-cellfunctions.Takentogether,ourdatademonstrateforthefirsttimethatseveralimmunopathologicalfactorsparticipateinthedownregulationofT-cellfunctionsinlocalPF.","container-title":"CellularandMolecularImmunology","DOI":"10.1038/cmi.2010.80","ISSN":"1672-7681","issue":"2","journalAbbreviation":"CellMolImmunol","note":"PMID:21258363\nPMCID:PMC4003141","page":"172-180","source":"PubMedCentral","title":"PleuralfluidfromtuberculouspleurisyinhibitsthefunctionsofTcellsandthedifferentiationofTh1cellsviaimmunosuppressivefactors","volume":"8","author":[{"family":"Li","given":"Qin"},{"family":"Li","given":"Li"},{"family":"Liu","given":"Yun"},{"family":"Fu","given":"Xiaoying"},{"family":"Qiao","given":"Dan"},{"family":"Wang","given":"Hui"},{"family":"Lao","given":"Suihua"},{"family":"Huang","given":"Fengyu"},{"family":"Wu","given":"Changyou"}],"issued":{"date-parts":[["2011",3]]}}},{"id":72,"uris":["/users/17061017/items/U9BIV8YL"],"itemData":{"id":72,"type":"article-journal","abstract":"Background\nTuberculouspleuraleffusion(TPE)isoneofthemostcommonformsofextrapulmonarytuberculosis(Tb).PatientswithTPEormalignantpleuraleffusions(MPE)frequentlyhaveasimilarlymphocyticpleuralfluidprofile.SincetheetiologyofPEinvariousdiseasesisdifferent,identifyingthecellularcomponentsmayprovidediagnosticcluesforunderstandingthepathogenesis.\n\nObjective\nWedeterminedthefrequencyofThelper(Th)subtypesinthePEsfordifferentiationofTbandnon-Tbpatients.\n\nMethods\nThirtypatientswithTPE,30patientswithMPE,14patientswithempyema(EMP),and14patientswithparapneumoniceffusion(PPE)wereenrolledbetweenDecember2018andDecember2019.Five-milliliterfreshPEintubescontainingheparinasananticoagulantwasobtainedfrompatients.ThefrequenciesofCD4+IL-9+,CD4+IL-22+,CD+IL-17+,andregulatoryT-cellsCD4+CD25+FOXP3+(Treg)weredeterminedbyflowcytometry.\n\nResults\nTregcellshavealowerfrequencyinTPEpatients[4.2(0.362–17.24)]comparedwithnon-TPEpatients[26.3(3.349–76.93,p<0.0001)].ThefrequencyofCD4+IL-9+cellswassignificantlylowerinTPEpatients[3.67(0.87–47.83)]comparedwithnon-TPEgroups[13.05(1.67–61.45),p<0.0001].Onthecontrary,therewasnosignificantdifferenceinthefrequencyofCD4+IL-17+andCD4+IL-22+cellsbetweenTPEandnon-TPEpatients(p=0.906andp=0.2188).Receiver-operatorcurve(ROC)analysisdemonstratedthatCD4+CD25+FOXP3+Tcells[optimalcutoffvalue=13.6(%),sensitivity90%,specificity75.86%]couldbeconsideredaspredictorforTPE.However,adenosinedeaminase[cutoffvalue27.5(IU/l),sensitivity90%,specificity96.5%]levelshadanevengreaterpredictivecapacity.\n\nConclusion\nADA,Tregcells,andCD4+IL-9+cellsmaydifferentiateTPEfromnon-TPEpatients.However,theseresultsneedvalidationinanindependentlargecohort.","container-title":"FrontiersinImmunology","DOI":"10.3389/fimmu.2021.780453","ISSN":"1664-3224","journalAbbreviation":"FrontImmunol","note":"PMID:34925358\nPMCID:PMC8674472","page":"780453","source":"PubMedCentral","title":"THelperCellSubsetsinthePleuralFluidofTuberculousPatientsDifferentiatePatientsWithNon-TuberculousPleuralEffusions","volume":"12","author":[{"family":"Roofchayee","given":"NedaDalil"},{"family":"Adcock","given":"IanM."},{"family":"Marjani","given":"Majid"},{"family":"Dezfuli","given":"NedaK."},{"family":"Varahram","given":"Mohammad"},{"family":"Garssen","given":"Johan"},{"family":"Mortaz","given":"Esmaeil"}],"issued":{"date-parts":[["2021",12,2]]}}}],"schema":"/citation-style-language/schema/raw/master/csl-citation.json"}[8,9]。cGVHD患者一般会表现出免疫重建的异常状态，就如T细胞亚群的不均衡、调节性T细胞（Treg）功能的缺失以及B细胞的异常活化ADDINZOTERO_ITEMCSL_CITATION{"citationID":"CfZWWJKd","properties":{"unsorted":true,"formattedCitation":"\\super[10,11]\\nosupersub{}","plainCitation":"[10,11]","noteIndex":0},"citationItems":[{"id":75,"uris":["/users/17061017/items/F6DZP2QD"],"itemData":{"id":75,"type":"article-journal","abstract":"目的\n探讨青蒿琥酯抗慢性移植物抗宿主病（cGVHD）的作用及可能机制。\n\n方法\n将B10D2小鼠的骨髓细胞和脾脏细胞混合悬液通过尾静脉输入近交系雌性BALB/c小鼠体内建立cGVHD模型，分别予青蒿琥酯（实验组）和丙酮腹腔（对照组）注射治疗。观察两组小鼠的cGVHD临床表现、生存时间和组织病理学改变；应用流式细胞术分析小鼠外周血和脾脏Th17和Treg细胞比例；免疫磁珠分选BALB/c小鼠脾脏CD4+T细胞，分别予丙酮和青蒿琥酯进行干预72h，流式细胞术分析两组Th17/Treg细胞比例。\n\n结果\n①实验组小鼠cGVHD临床症状较对照组明显减轻，生存时间延长[（55.71±6.99）d对（46.57±7.83）d，χ2=5.457，P=0.020]。②实验组小鼠皮肤及肺脏cGVHD病理损伤较轻。③与对照组比较，实验组小鼠外周血、脾脏Th17细胞比例均降低[（0.58±0.19）%对（1.51±0.18）%，t=7.233，P<0.001；（0.71±0.18）%对（1.48±0.38）%，t=3.653，P=0.011]，Treg细胞比例增高[（8.40±0.23）%对（4.45±0.04）%，t=15.680，P<0.001；（10.48±0.48）%对（6.62±0.24）%，t=6.590，P=0.003]，Th17/Treg细胞比值均下降（0.09±0.03对0.34±0.05，t=7.621，P=0.002；0.06±0.02对0.19±0.03，t=6.993，P=0.002）。④体外培养小鼠脾脏CD4+T细胞，青蒿琥酯干预组较对照组Th17细胞比例减低[（0.82±0.37）%对（3.39±1.22）%，t=4.044，P=0.007]，Treg细胞比例明显增高[（34.63±1.29）%对（14.28±1.69）%，t=19.119，P<0.001]，Th17/Treg细胞比值下降（0.24±0.09对0.02±0.01，t=4.780，P=0.003）。\n\n结论\n青蒿琥酯可通过减少Th17细胞及增加Treg细胞，恢复Th17/Treg平衡，减轻cGVHD的临床与病理学损伤，从而发挥抗cGVHD作用。","container-title":"ChineseJournalofHematology","DOI":"10.3760/cma.j.issn.0253-2727.2019.01.012","ISSN":"0253-2727","issue":"1","journalAbbreviation":"ZhonghuaXueYeXueZaZhi","note":"PMID:30704231\nPMCID:PMC7351699","page":"63-68","source":"PubMedCentral","title":"青蒿琥酯调控Th17/Treg平衡抑制慢性移植物抗宿主病的实验研究","volume":"40","issued":{"date-parts":[["2019",1]]}}},{"id":77,"uris":["/users/17061017/items/P9EEDLK5"],"itemData":{"id":77,"type":"article-journal","abstract":"Thesuccessofallogeneichematopoieticstemcelltransplantation(allo-HSCT)inthetreatmentofhematologicalmalignanciesremainshamperedbylife-threateningchronicgraftvs.hostdisease(cGVHD).Althoughmultifactorialinnature,cGVHDhasbeenassociatedwithimbalancesbetweeneffectorandregulatoryTcells(Treg).Tofurtherelucidatethisissue,weperformedaprospectiveanalysisofpatientsundergoingunrelateddonorallo-HSCTafterareducedintensityconditioning(RIC)regimencontaininganti-thymocyteglobulin(ATG)andthesameGVHDprophylaxis,atasingleinstitution.WestudiedTcellsubsethomeostasisovera24-monthfollow-upafterHSCTinacomparativeanalysisofpatientswithandwithoutcGVHD.WealsoquantifiednaiveandmemoryTcellsubsets,proliferationandexpressionoftheapoptosis-relatedproteinsBcl-2andCD95.Finally,weassessedthymicfunctionbyTcellreceptorexcisioncircle(TREC)quantificationandTcellreceptor(TCR)diversitybyTCRVβspectratyping.WhilethetotalnumberofconventionalCD4(Tcon)andCD8Tcellswassimilarbetweenpatientgroups,TregweredecreasedincGVHDpatients.Interestingly,wealsoobserveddivergentpatternsofNaiveandStemCellMemory(SCM)subsetrecoveryinTregandTconcomparedtoCD8.PatientswithcGVHDshowedimpairedrecoveryofNaiveandSCMTconandTreg,butsignificantlyincreasedfrequenciesandabsolutenumbersofNaiveandSCMwereobservedintheCD8pool.MarkedlyincreasedEMRACD8TcellswerealsonotedincGVHD.Takentogether,theseresultssuggestthatNaive,SCMandEMRACD8playaroleintheemergenceofcGHVD.ReducedNaiveandrecentthymicemigrantTconandTregincGVHDwaslikelyduetoimpairedthymicoutput,asitwasaccompaniedbydecreasedCD4TRECandTCRdiversity.Ontheotherhand,CD8TCRdiversitywassimilarbetweenpatientgroups.Furthermore,nocorrelationwasobservedbetweenCD8TRECcontentandNaiveCD8numbers,suggestinglimitedthymicproductionofNaiveCD8Tcellsinpatientsaftertransplant,especiallyinthosedevelopingcGVHD.ThemechanismsbehindtheopposingpatternsofCD4andCD8subsetcellrecoveryincGVHDremainelusive,butmaybelinkedtothymicdamageassociatedwiththeconditioningregimenand/oracuteGVHD.","container-title":"FrontiersinImmunology","DOI":"10.3389/fimmu.2019.00334","ISSN":"1664-3224","journalAbbreviation":"FrontImmunol","note":"PMID:30894856\nPMCID:PMC6414429","page":"334","source":"PubMedCentral","title":"NaiveandStemCellMemoryTCellSubsetRecoveryRevealsOpposingReconstitutionPatternsinCD4andCD8TCellsinChronicGraftvs.HostDisease","volume":"10","author":[{"family":"Soares","given":"MariaV."},{"family":"Azevedo","given":"RitaI."},{"family":"Ferreira","given":"InêsA."},{"family":"Bucar","given":"Sara"},{"family":"Ribeiro","given":"AnaC."},{"family":"Vieira","given":"Ana"},{"family":"Pereira","given":"PauloN.G."},{"family":"Ribeiro","given":"RuyM."},{"family":"Ligeiro","given":"Dario"},{"family":"Alho","given":"AnaC."},{"family":"Soares","given":"