PCR-STR用于异基因造血干细胞移植后的骨髓和外周血嵌合状态的监测及作用

目的用PCR-STR检测观察比较异基因造血干细胞移植术后,骨髓和外周血嵌合状态和植入状态。方法提取17例异基因造血干细胞移植术中的供者外周血及受者移植前后各阶段外周血和骨髓的DNA,PCR PCR-STR检测扩增15个基因位点,和1个性别位点AMEL。用遗传分析仪进行毛细管电泳,确定基因位点,根据基因型的差异选择嵌合率计算公式,分析植入情况和嵌合状态。结果12名患者完全植入,骨髓和外周血的PCR-STR结果一致;1名患者骨髓PCR-STR显示为持续未植入,外周血PCR-STR显示为短期混合植入;4名患者骨髓和外周血PCR-STR显示嵌合状态时间不一致,骨髓PCR-STR显示嵌合状态时间明显早于外周血(P<0.05)。结论PCR-STR是分析异基因造血干细胞移植后供体是否植入的灵敏、准确度高的方法,骨髓嵌合状态的变化的出现早于外周血,混合嵌合状态对白血病复发有预警作用,骨髓嵌合状态的早期变化对实施临床干预治疗尤为重要。     

异基因造血干细胞移植侵袭性真菌感染26例临床分析

目的 探讨异基因造血干细胞移植患者发生侵袭性真菌感染的高危因素、临床特点、治疗和预后.方法 分析我科2002年3月至2010年7月行造血干细胞移植术100例患者发生侵袭性真菌感染26例患者的临床资料.造血干细胞移植前发生肺部真菌感染7例;造血干细胞移植后发生肺部真菌感染14例,呼吸道真菌感染3例,肠道真菌感染2例.观察侵袭性真菌感染患者的移植物抗宿主病的发生、巨细胞病毒血症发生、淋巴细胞亚群检测、合并其他慢性基础疾病情况.将侵袭性真菌感染患者分为2组,拟诊经验性治疗组12例及临床诊断抢先治疗组14例.结果 26例侵袭性真菌感染的患者合并移植物抗宿主病20例,合并巨细胞病毒血症6例,细胞免疫功能低下的19例.合并糖尿病5例,肺结核3例,支气管扩张1例.经验性治疗组完全治愈8例(67%),疾病进展1例(8%).抢先治疗组完全治愈3例(21%);疾病进展5例(36%),2组比较差异有统计学意义(χ2=5.418,P<0.05).结论 造血干细胞移植存在侵袭性真菌感染高危因素的患者更应引起临床医师高度的重视.

Abstract：

Objective To explore the high-risk factors,clinical characteristics,therapy and prognosis of invasive fungal infection (IFI)in patients underwent allogeneic haemopoietic stem cell transplantation (AlloHSCT). Methods One hundred patients underwent Allo-HSCT at our department from March 2002 to July 2010 were analyzed retrospectively,among whom 26 patients had invasive fungal infection(IFI). Seven patients had pulmonary IFI before allo-HSCT, 14 patients had pulmonary IFI after allo-HSCT,3 patients had respiratory tract system IFI, and 2 patients had intestinal IFI. We observed the occurrence of Graft-versus-host disease (GVHD) ,cytomegalovirus( CMV )infection, Lymphocyte subsets and chronic basic diseases in patients with IFI. The twenty six cases were divided into two groups: experience therapy group with 12 cases and preemption therapy group with 14 cases. Results Among 26 patients with IFI,20 cases suffered from GVHD,6 cases had CMV infection,19 cases had low cellular immune function simultaneously. 1 case had diabetes,3 patients had pulmonary tuberculosis and 1 case had bronchiectasis as complications. In experience therapy groupe: 8 cases (67%)recovered completely but 1 case(8% )suffered from progressive infection. In preemption therapy groupe:3 cases ( 21% ) recovered completely but 5 cases ( 36% ) suffered from progressive infection. Conclusion Clinician should pay close attention to the patients with high-risk factors of IFI after allo-HSCT.     

T否决细胞促进造血干细胞植入的研究

否决效应是一种能特异性抑制识别否决细胞自身表面抗原的细胞毒性T细胞前体细胞(CTL-p)的攻击,而CTL-p对识别第三方抗原无抑制作用。具有否决效应的细胞称为否决细胞。细胞毒性T淋巴细胞(cytotoxic Tlymphocyte,CD8+CTL)是现知否决活性最强的细胞。在异基因造血干细胞移植中,输注供者源的CD8+CTL否决细胞清除宿主同种异体反应细胞可以促进供者干细胞的植入。本文就近年来关于CD8+CTL否决效应的机制、GVH效应、抗肿瘤效应、活体内的应用及与药物和细胞之间的协同作用的研究进展做一综述。     

The experience of allogeneic hematopoietic stem cell transplantation in a patient with X-linked adrenoleukodystrophy

X-linked adrenoleukodystrophy (X-ALD), a progressive neurometabolic disorder that is caused by a defect in the gene ABCD1 (ATP-binding cassette, subfamily D, member 1), which encodes the peroxisomal ABC half-transporter ALD protein. Recently, allogeneic hematopoietic stem cell transplantation (alloHSCT) is the only therapy known to prevent disease progression. In this study, we would like to present our experience of alloHSCT for X-ALD from a HLA matched related sibling by the use of reduced intensity conditioning regimen composed of fludarabine, busulfan and ATG which allows us to reduce procedure-related toxicity and prevent mortality while achieving a curative effect.     

异基因造血干细胞移植后并发自身免疫性溶血性贫血患者产生类抗体血清学检测方法及输血策略

目的探讨异基因造血干细胞移植（AHSCT）后免疫介导的自身免疫性溶血性贫血（AIHA）患者交叉配血不合的原因,血浆中不规则抗体筛选试验阳性时,不规则抗体鉴定的血清学检测方法的选择,处理措施及输血策略。 方法一例AHSCT后的3岁男性患儿因重度贫血为改善贫血症状需输注红细胞入住四川省人民医院。通过盐水试管法确定了患儿ABO、Rh血型后进行交叉配血试验发现患儿与多个同型供者血液不相合,考虑到患儿贫血严重,于是采用直接抗球蛋白试验来判断红细胞是否被致敏;通过盐水介质试管法、微柱抗球蛋白法和聚凝胺法进行ABO血型系统以外的不规则抗体筛选试验以确定血浆中有无不规则抗体;根据不规则抗体筛选试验结果选择聚凝胺法来确定抗体的特异性;通过盐水介质试管法、经典抗球蛋白法和聚凝胺法进行交叉配血实验,结合抗体鉴定的结果,综合分析选择合适的供者红细胞输注。 结果本例患儿ABO血型为AB型,Rh分型为CCDee,ABO血型已转变为供者血型。直接抗球蛋白试验强阳性,红细胞被抗体致敏。血浆中检出了不规则抗体,红细胞上放散下来的致敏抗体与血清中检出的不规则抗体均为类抗-Ce自身抗体。选择了避开类抗-Ce抗体的Ce抗原阴性的AB型红细胞输注后血色素升高,3 d后复查血常规Hb为79 g/L,输血有效。 结论任何类型的AHSCT后都有可能发展成AIHA,也就是血浆中可能存在某种自身抗体（类抗体）而破坏自身红细胞,若能够明确患者血清中存在的类抗体,即可避开这种抗体而筛选红细胞无相应抗原的供者,也就避免发生溶血性输血反应以安全输血。     

Successful collection and engraftment of autologous peripheral blood progenitor cells in poorly mobilized patients receiving high-dose granulocyte colony-stimulating factor

Background : Granulocyte Colony‐Stimulating Factor (G‐CSF) alone or in conjunction with chemotherapy is commonly used to mobilize hematopoietic progenitor cells (HPC) into peripheral blood for progenitor cell harvest for autologous HPC transplantation. However, in up to 30% of patients, HPC are not effectively mobilized. In this study, we report the efficacy and safety profiles of a mobilization strategy using high‐dose (up to 36 μg/kg) G‐CSF in poorly mobilized patients. Study Design and Methods : Retrospective medical record reviews were performed for 392 patients who underwent autologous peripheral blood progenitor cell collection. A total of 56 patients were given high‐dose G‐CSF due to very ineffective mobilization and 35 of these patients underwent autologous HPC transplantation. The efficacy of mobilization, apheresis collection, and infusion were reviewed and analyzed. Results : More than 2.5 × 10⁶ CD34/Kg were collected in 88% of patients (49 of 56) who were placed on high‐dose G‐CSF due to very ineffective mobilization. Of the 35 patients who underwent HPC transplantation using the progenitor cells that were mobilized with high‐dose G‐CSF due to very ineffective mobilization, all had rapid and complete neutrophil and platelet engraftment comparable with good mobilizers. Conclusion : We conclude that collection of HPC using hyperstimulation with G‐CSF is an effective alternative approach for HPC harvest for poorly mobilized patients. J. Clin. Apheresis 27:235–241, 2012. © 2012 Wiley Periodicals, Inc.     

Peripheral blood stem cell grafts in allogeneic hematopoietic cell transplantation: It is not all about the CD34+ cell dose

Allogeneic Hematopoietic Cell Transplantation is a curative approach in various malignant and non-malignant disorders. The majority of adult transplants in the current era are performed using mobilized stem cells, harvested from the peripheral blood by leukapheresis. Peripheral blood stem cell (PBSC) collections are designed to target a dose of stem cells that will result in safe engraftment and hematopoietic recovery; however, 99 % of the cells contained in a PBSC graft are not stem cells and a growing number of studies attempt to characterize the associations between graft composition and transplant outcomes. A better understanding of the impact of the quantity and quality of various cell types in PBSC grafts may lead to development of novel collection strategies or improved donor selection algorithms. Here we review relevant findings from recent studies in this area.     

Hematopoietic stem cell transplantation in geriatric patients in Turkey

The incidence of most hematologic malignancies increases with age. Physicians increasingly refer older patients for hematopoietic stem cell transplantation (HSCT) due to more experience and improved supportive care in HSCT. This article discusses the available data regarding the feasibility, tolerability, toxicity, and effectiveness of autologous and allogeneic HSCT in older adults.     

异基因造血干细胞移植后伴噬血现象分析

目的分析异基因造血干细胞移植(allo-HSCT)后伴噬血现象的特征。方法选取上海市第一人民医院行allo-HSCT后出现噬血细胞性淋巴组织细胞增多症(HLH)的患者2例,讨论其可能的病因。结果患者一入院诊断为免疫缺陷病,行allo-HSCT 13 d后粒系植入,血红蛋白、血小板始终偏低,凝血功能差,消化道出血,巨细胞病毒性肠炎,肺部感染,骨髓细胞形态学检查可见噬血现象。患者二入院诊断为急性单核细胞性白血病,行allo-HSCT 14 d后粒系植入,行allo-HSCT 42 d后三系降低,发热,铁蛋白高,骨髓细胞形态学检查可见噬血现象,嵌合率进行性降低。结论allo-HSCT后伴噬血现象非常罕见,且进展迅速,预后不良,噬血现象常因感染所致。当行allo-HSCT后骨髓出现噬血现象时,应积极排查感染,检测原发病是否复发,同时应检测患者及其父母相关基因,尽早治疗,争取取得更好的预后。     

Mycobacterium abscessus and massiliense lung infection during macrolide treatment for bronchiolitis obliterans after allogeneic hematopoietic stem cell transplantation

In patients undergoing allogeneic hematopoietic stem cell transplantation (allo-SCT), post-transplant lung infection is critical for their prognosis. Mycobacterium abscessus complex is not fully recognized as a nontuberculous mycobacteria (NTM) pathogen of post-SCT lung infection. Here, we present three post-allogeneic SCT patients who developed pulmonary infection caused by M. abscessus complex including M. abscessus and M. massiliense. In all three cases, macrolide antibiotics had been administered for bronchiolitis obliterans syndrome (BOS) before the confirmation of their infection, and macrolide resistance was noted in the M. abscessus isolates, one of which resulted in an unfavorable treatment outcome. It is important to consider M. abscessus lung infection as well as other NTM in patients receiving allo-SCT, particularly those receiving macrolide therapy for BOS.     

实时定量聚合酶链反应检测巨细胞病毒在同种异基因造血干细胞移植术后监测中的意义

目的:探讨同种异基因造血干细胞移植(allo-HSCT)中巨细胞病毒(cytomegalovirus,CMV)激活情况及激活相关因素。方法:52例接受allo-HSCT的患者为研究对象,所有患者及其供者移植前均为CMV携带者,即为CMV血清型阳性。所有移植术后患者应用实时定量聚合酶链反应(PCR)方法对CMVDNA进行定量检测。结果:移植后100d内,52例患者中28例检测到CMVDNA复制。造血干细胞移植(HSCT)供者类型和移植物抗宿主反应(graft versus host disease,GVHD)的发生与CMV激活明显相关(P值分别为0.018和0.039)。在44例可评估患者中,allo-HSCT100d后仅7例检测到CMVDNA复制,其发生率与移植后发生GVHD且接受激素治疗(P=0.0009)、人类白细胞抗原(HLA)全相合无关供者移植或者HLA部分相合同胞供者移植有关(P=0.037)。应用实时定量PCR检测发现,CMV对更昔洛韦抢先治疗的应答存在2种动力学模式(快速型和缓慢型),其中表现为缓慢型的患者CMV最高负载量较快速型高10倍以上,其病毒衰减时间也要延长3～4周,多数发生在因出现GVHD而接受激素治疗以及接受HLA全相合无关供者和HLA部分相合同胞供者干细胞移植的患者。结论:移植前CMV血清型阳性患者接受allo-HSCT后100d内出现CMV激活是常见并发症。移植100d之后,对于未发生GVHD而未使用激素治疗者或以HLA全相合同胞为供体的患者,CMV激活概率低,无需进行常规频繁监测。     

脐血造血干细胞移植中脐血输注的护理

脐血造血干细胞移植中如何使脐血造血干细胞顺利输注人病人体内是移植成功的重要环节,因此,脐血输注的护理尤为重要。笔阐述9例脐血造血干细胞移植患,脐血输注前输注时的护理,脐血输注后并发症的观察及护理3个部分的护理经验。9例患均成功进行了脐血输注。     

Treatment with granulocyte colony-stimulating factor increases interleukin-1 receptor antagonist levels during engraftment following allogeneic stem-cell transplantation

BACKGROUND: The effect of treatment with granulocyte colony-stimulating factor (G-CSF) on interleukin-1 receptor antagonist (IL-1ra) plasma concentrations as well as the role of IL-1ra on leucocyte recovery and parameters of infection within the first 30 days after haematopietic stem-cell transplantation (HSCT) are not well known. MATERIAL AND METHODS: Twenty-seven patients undergoing myeloablative therapy followed by allogeneic SCT for various haematological disorders were either treated with (n = 18) or without (n = 9) G-CSF. IL-1ra plasma levels were serially determined by ELISA starting at day - 1 and continued until patients were engrafted. RESULTS: Patients receiving G-CSF had significantly shorter neutropenic periods and significantly lower mean C-reactive protein serum levels during the first 3 weeks succeeding bone marrow transplantation (BMT). Importantly, starting at day + 11 and paralleling the rise of peripheral blood leucocytes, increasing IL-1ra plasma concentrations were observed in both treatment groups. However, the magnitude of the IL-1ra surge was far greater in the G-CSF treatment group. Peak IL-1ra plasma level observed on day + 19 was 882.3 +/- 879.2 pg mL(-1) (mean +/- SD) in patients receiving G-CSF compared with 285.8 +/- 175.2 pg mL(-1) (mean +/- SD) in patients not receiving G-CSF (P = 0.0130). Furthermore, a direct correlation of IL-1ra with peripheral blood leucocytes was verified by the Spearman rank test (P = 0.0025). CONCLUSION: Granulocyte colony-stimulating factor-mediated acceleration of neutrophil recovery following myeloablative therapy correlated with increased IL-1ra plasma concentrations. Our data suggest that IL-1ra constitutes an intrinsic component of the anti-inflammatory and neutrophil differentiating efficacy of G-CSF and, thus, IL-1ra may be required for the in vivo activity of G-CSF.     

Surgical resection for persistent localized pulmonary fungal infection prior to allogeneic hematopoietic stem cell transplantation: Analysis of six cases

ObjectiveAlthough invasive fungal disease (IFD) is an important complication in allogeneic hematopoietic stem cell transplantation (HSCT), the clinical significance of surgery, including the role of surgical resection for persistent pulmonary fungal disease prior to allogeneic HSCT in the current era with a variety of available antifungal agents, is controversial. We investigated the role of surgical resection.MethodsWe retrospectively investigated six patients who underwent surgical resection of suspected pulmonary fungal disease prior to allogeneic HSCT between April 2007 and June 2016 at our medical center.ResultsWe present six patients who underwent surgical resection of suspected pulmonary fungal disease prior to allogeneic HSCT. In our case series, three of four patients who were given a presurgical diagnosis of possible IFD were given a proven diagnosis after surgery, including two cases of invasive aspergillosis (IA) and one case of mucormycosis. All surgeries were performed by video-assisted thoracic surgery (VATS) for lobectomy without major complications. Recurrence of IFD was not observed after allogeneic HSCT in any of the six patients.ConclusionOur experience indicated that surgical resection of persistent localized pulmonary lesions of IFD before allogeneic HSCT was helpful for obtaining a definitive diagnosis and might be useful for reducing recurrence after HSCT.     

异基因造血干细胞移植后供者外周血造血干细胞输注预防高危白血病复发

目的评估重组人粒细胞集落刺激因子(G-CSF)动员的供者外周血造血干细胞输注 (GPBSCI)作为一种早期过继性免疫治疗方法的有效性和安全性。方法 12例高危白血病患者同胞配型相合异基因造血干细胞移植(allo-HSCT)后接受了G-CSF动员的预防性GPBSCI。allo-HSCT前患者的诊断包括2例Ph 急性淋巴细胞白血病首次完全缓解(ALL-CR1),1例ALL-CR2,1例ALL合并顽固中枢神经系统白血病(CNSL),1例急性髓系白血病(AML)复发,1例AML合并CNSL,1例 AML-CR3,4例进展期慢性粒细胞白血病(CML)及1例骨髓增生异常综合征-难治性贫血伴幼稚细胞增多型(MDS-RAEB)。结果 12例患者共接受了16次GPBSCI,其中移植后90天( 90天)前接受 GPBSCI 5次,输注的单个核细胞(MNC)及CD3 细胞中位数分别为1．00(0．95-1．24)×108/kg和 0．53(0．39-0．63)×108/kg。 90天后接受GPBSCI 11次,输注两类细胞中位数分别为2．27(1．00- 4．30)×108/kg和1．15(0．55-2．10)×108/kg。输注后4例患者发生了Ⅰ或Ⅱ度急性移植物抗宿主病(GVHD),1例患者发生Ⅲ度急性GVHD。7例患者发生了慢性GVHD,其中4例为广泛型。2例患者未发生输注相关GVHD。未观察到GPBSCI相关的全血细胞减少。中位随访563(415-728)d,12 例高危白血病患者中有10例无病存活,2例死于白血病复发。结论预防性GPBSCI可以增强移植物抗白血病作用,相关不良反应小,可能成为改善高危白血病allo-HSCT预后的安全有效的手段。     

ABO血型对异基因造血干细胞移植患者植入的影响探讨

目的 探讨ABO同型及3种ABO血型不合对异基因造血干细胞移植患者各系植入的影响,为患者优化移植方案提供依据.方法 回顾性分析本院2014年1月～2018年6月期间进行异基因造血干细胞移植患者70例,ABO同型18例,ABO血型不合52例,在确保对比组患者年龄;性别;供/受者亲缘关系;疾病诊断;移植前骨髓造血功能状况;...     

HLA相合同胞脐血移植成功治疗一例成人慢性髓系白血病

目的研究脐血移植(CBT)治疗成人恶性血液病的可行性,观察长期造血重建和移植相关并发症的发生.方法 1例18岁体重75 kg的慢性髓系白血病慢性期(CML-CP)患者在用改良的马利兰、环磷酰胺(Bu/CTX)方案预处理后给予HLA相合同胞CBT.输入有核细胞数1.73×107/kg,CD34+细胞为2.7×105/kg.移植物抗宿主病(GVHD)预防方案选用环孢菌素A(CsA)加甲泼尼龙.结果移植后第18天中性粒细胞计数>0.5×109/L,移植后36天血小板计数>50×109/L.CBT后共给予7次血小板输注.移植后80天DNA位点检测示已全部转为供者型.移植后90天后出现严重黄疸,诊断为肝脏急性GVHD合并巨细胞病毒感染,通过加用免疫抑制剂和抗病毒药物治疗,同时辅以血浆交换和人工肝体外胆红素吸附,并发症得到控制.随访24个月,患者一般情况良好,肝功能基本正常,骨髓细胞Ph染色体和bcr/abl基因持续阴性.结论本例为国内首例应用异基因CBT成功治疗成人白血病,表明异基因CBT应用于成人血液病的治疗是可行的.