异基因外周血造血干细胞移植后ABO血型抗原抗体效价的变化

目的:观察ABO血型不合的患者异基因造血干细胞移植前后血型抗原转变及抗体效价的变化,为患者输注血液成分的选择提供依据。方法:在干细胞移植前对患者和供者进行HLA配型、ABO及RhD血型鉴定,移植后定期检测患者ABO血型抗原强度及抗体效价的变化,对输注血液成分的种类进行统计分析。结果:22例ABO血型不合的患者干细胞移植后28～45 d血型抗原转变为供者血型,56～80 d血清中可检出凝集素,抗体效价为1∶4～1∶8,但部分患者的血清中未检出相应的凝集素,正反定型结果不相符。结论:ABO血型不合的干细胞移植后,患者血型抗原转变为供者血型,输血时应选择不被患者血清中抗体所凝集的不含相应抗原的红细胞,以及供者血清中不含相应抗体的血小板或血浆进行输注,确保输血安全。     

异基因造血干细胞移植后免疫功能重建

异基因造血干细胞移植(allo-HSCT)后患者的免疫功能受到严重损伤,移植后免疫功能低下主要表现在①免疫个体发生学过程再现的削弱;②缺乏长期稳定的供体免疫功能的转移;③移植物抗宿主病(GVHD)对免疫功能的影响;④胸腺功能的降低.临床实践证实,清髓性治疗后患者的免疫功能处于麻痹状态可以持续1年之久.     

清肿瘤性异基因造血干细胞移植

标准的清髓性异基因造血干细胞移植(allo-HSCT)对于需代替治疗的造血与免疫系统的非恶性疾病,应当是合理或足够的;然而,对于恶性血液病患者,清除患者骨髓造血组织,成功重建异体正常造血与免疫系统,并不一定能完全治愈恶性血液病,因为白血病(干)细胞并非只限骨髓中存在,它可浸润骨髓之外的其他任何组织。临床实践证实,allo-HSCT后仍然有30%左右的患者疾病复发,特别是具有高危因素或难治复发患者复发率可高达40%～70%以上。这些复发的白血病细胞几乎全系源自患者移植前本身的白血病细胞,其中半数患者以髓外部位复发开始,有证据提示,清髓性移植并没有完全杀灭患者体内的白血病细胞,特别是那些对化放疗不敏感或栖居在髓外"庇护所"中的白血病干细胞,最终导致疾病复发。因此笔者提出并建立了一个清肿瘤性异体造血干细胞移植(TAHSCT)的概念,在临床上对其进行了初步的探讨。其内容贯穿于移植技术全过程的各个环节,但主要为应用个体化清肿瘤性预处理方案和加强移植后免疫治疗。     

异基因造血干细胞移植治疗恶性血液病临床疗效分析

目的总结162例异基因造血干细胞移植（allo-HSCT）[包括：同胞allo-HSCT125例．非血缘关系allo-HSCT30例和非清髓异基因外周血造血干细胞移植（allo-PBSCT）7例1治疗恶性血液病疗效和生存状况。方法慢性粒细胞白血病（CML）患者62例，急性髓系白血病（AML）患者58例，急性淋巴细胞白血病（ALL）患者28例．骨髓增生异常综合征（MDS）6例，多发性骨髓瘤（MM）3例，非霍奇金淋巴瘤（NHL）3例，慢性粒单细胞性白血病（CMML）1例．霍奇金淋巴瘤（HD）1例。经预处理，进行人类白细胞抗原（HLA）相合的同胞异基因骨髓移植（allo-BMT）27例．allo-PBSCT98例和非血缘关系allo-BMT4例，非血缘关系aUo-PBSCT26例．HLA相合的同胞非清髓allo-PBSCT7例。非血缘关系allo-HSCT患者采用长程加强的移植物抗宿主病（GVHD）的预防方案（将环孢菌素A提前至预处理开始时使用，同时加用霉酚酸酯）。结果移植后中位随访38（2-259）个月。allo-HSCT患者长期DFS为54、9％（89／162），CML至今DFS为61-3％（38／62），AML至今DFS为56，9％（33／58），ALL至今DFS为39-3％（11／28）．MDS至今DFS为83-3％（5／6），3例NHL存活1例，1例CMML存活．3例MM均死亡．霍奇金淋巴瘤（HD）1例死亡。移植后100d内移植相关死亡率（TRM）19．8％（32／162），死亡原因分别为急性GVHD、播散性感染、复发和植入失败；移植后100d至2年内TRM为24、7％（40／162）．死亡原因分别为慢性GVHD、CMV感染和疾病复发，1例于移植后1413d死亡．其余移植超过2年均存活，死亡原因是慢性GVHD合并感染，最长生存已11年。结论allo-HSCT可使相当部分白血病患者获得长期无病存活．治疗MDS效果良好．治疗MM效果很差。该组患者移植后100d内死亡原因主要是急性GVHD：移植后100d至2年内死亡的主要原因是慢性GVHD和CMV感染、疾病复发；故除正确处理移植相关并发症?     

异基因造血干细胞移植后的移植物抗白血病作用的临床观察

移植物抗白血病作用（GVL）是异基因造血干细胞移植后,引起造血系统肿瘤持续缓解状态的一种免疫介导的反应。目前,对GVL的机理仍不清楚。临床上观察到,在GVL出现前,病人多经历了一个急或慢性的移植物抗宿主病（GVHD）过程。GVHD使受者正常的组织和细胞受到损伤,而同时产生     

异基因造血干细胞移植后骨髓免疫微环境重建研究

GVHD 和GVL 是影响异基因造血干细胞移植术成败的关键因素。骨髓不仅是造血器官,更是免疫器官,骨髓中的免疫微环境关乎移植后造血重建、免疫功能重建、移植后白血病复发。本文将从骨髓微环境的病理生理特点、异基因造血干细胞移植后免疫功能重建、骨髓免疫微环境与移植后造血恢复及移植后白血病复发等角度,综述近年来关于异基因造血干细胞移植后骨髓免疫微环境重建与造血重建和白血病复发相关研究情况。     

pp65抗原血症监测异基因造血干细胞移植受体CMV的感染

人巨细胞病毒（cytomegalovirus，CMV）感染是异基因造血干细胞移植（Allo-HSCT）术后重要并发症和首位感染性死亡原因。若不加以预防或治疗，12％～20％的Allo-HSCT患者术后会发生严重的CMV感染，病死率〉85％。     

异基因造血干细胞移植后乙型肝炎病毒感染2例临床干预并文献复习

目的加强对造血干细胞移植中HBV感染的重视,注重早期干预,提高移植成功率。方法回顾性分析97例异基因造血干细胞移植患儿的临床资料,通过对2例HBV感染的诊治体会结合文献复习。结果2001年5月至2008年5月在上海交通大学附属上海儿童医学中心接受异基因造血于细胞移植的97例患儿中,2例分别在移植后41d（病例1）、15个月（病例2）发生HBV感染。病例1移植前肝功能正常,乙肝二对半检查阴性,回顾性分析发现该患儿移植时HBV正处于潜伏状态（HBV—DNA1．17×10^6copies·mL^-1）。该患儿乙肝来势凶猛,移植后41～43d出现巩膜明显黄染并伴大量腹水,移植后46d迅速发展至肝、肾功能衰竭,出现少尿,凝血酶原时间38．4S,部分凝血酶原时间〉120S,凝血酶时间〉100s,Cr251μmol·L^-1,ALT3195U·L^-1,血清总胆红素7mg·L^-1,直接鹏红素2．8mg·L^-1,HBV—DNA1,08×10^3copies·mL^-1,经拉米夫定等积极治疗2周后好转。移植后130d随着移植物抗宿主病（GVHD）的复燃和免疫抑制药物的加强应用,HBV再度活跃,HBV—DNA从原已控制的3．50×10^4copies·mL^-1逐升至2．05×10^6copies·mL^-1,移植后315d出现HBVYMDD（＋）变异株,遂予阿德福韦酯联合治疗至今（移植后3．5年）,目前肝、肾功能正常。病例2白血病起病初及干细胞移植前均示HBs-Ab（＋）、HBc—Ab（＋）、HBe—Ab（＋）,A¨和HBV—DNA正常,移植后12个月发生慢性广泛性GVHD,加强抗排异治疗后于移植后15个月复查发现：ALT168U·L^-1,HBV-DNA升至5×10^8copies·mL^-1,出现HBs-Ag（＋）和HBe-Ag（＋）。予拉米夫定治疗至移植后4．5年,目前ALT40～80U·L^-1,HBV—DNA1×10^3～1×10^4copies·mL^-1。结论乙肝在移植患儿巾并不少见,长期的免疫抑制治疗常使病情反复,加强病毒监测、重视早期干预至关重要;移植前HBVDNA检测有助于发现潜伏期患儿;HBs—Ab（＋）、HBe—Ab（＋）和HBc—Ab（＋）患儿在强烈免疫抑制下仍有HBV复燃的风险。     

异基因造血干细胞移植后免疫功能重建

异基因造血干细胞移植(allo-HSCT)，岳患者的免疫功能受到严重损伤，移植后免疫功能低下主要表现在：①免疫个体发生学过程再现的削弱；②缺乏长期稳定的供体免疫功能的转移； ③移植物抗宿主病(GVHD)对免疫功能的影响； ④胸腺功能的降低。临床实践证实，清髓性治疗后患者的免疫功能处于麻痹状态可以持续1年之久。     

异基因造血干细胞移植后NK细胞免疫重建的研究进展

异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation, allo-HSCT)是治疗血液系统恶性肿瘤有效且唯一的治愈方法,移植成功与否主要取决于移植后受者造血系统和免疫系统的恢复程度。HSCT后免疫系统的恢复,也被称为免疫重建,其中NK细胞的免疫重建最为重要,它在个体的早期防御和allo-HSCT的成败中发挥重要的作用。文章即对allo-HSCT中NK细胞免疫重建规律、促进NK细胞免疫重建的相关免疫治疗方法及疗效的进展作一综述。     

单倍型异基因造血干细胞移植后发生巨细胞病毒感染的临床分析

目的 探讨单倍型异基因造血干细胞移植后发生巨细胞病毒(CMV)感染的临床特点及治疗.方法 对北京军区总医院血液科2011年1月-2013年1月采用单倍型异基因造血干细胞移植治疗的50例患者的临床资料进行回顾性研究,供者接受粒细胞集落刺激因子动员,采用骨髓加外周血干细胞联合移植,预处理方案以改良BUCY方案为主,移植后监测CMV-DNA,分析CMV感染的特点并探讨其治疗方法.结果 全部患者中男32例,女18例,年龄5-45岁,平均年龄23.8岁,其中共12例发生CMV血症,发生率为24％,首次检出CMV-DNA中位时间是移植后50天(36-72),CMV定量范围为2.4×103-6.2×106拷贝/mL,其中6例为CMV相关性出血性膀胱炎,2例为CMV相关性间质性肺炎,抗CMV治疗后全部患者CMV-DNA转阴.结论 单倍型异基因造血干细胞移植后巨细胞病毒感染发生率高,更昔洛韦、膦甲酸钠抗病毒治疗疗效可靠、不良反应少.     

Cytomegalovirus management after allogeneic hematopoietic stem cell transplantation: A mini-review

Because of the high incidence of cytomegalovirus (CMV) seropositivity in the population, CMV infection is a common and severe complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Taiwan. Here we propose a CMV management strategy for patients undergoing allo-HSCT from the Taiwanese perspective, which focuses on the epidemiology, diagnosis, monitoring, prophylaxis, and treatment of CMV infection after allo-HSCT. In terms of CMV monitoring, weekly CMV monitoring with the COBAS® AmpliPrep system is the standard approach because the pp65 CMV antigenemia assay has a lower sensitivity than CMV monitoring with the COBAS® AmpliPrep system. However, pp65 CMV antigenemia assay has a better correlation with clinical symptoms in immunocompromised patients. A 14-week prophylactic course of letermovir is recommended for allo-HSCT recipients in Taiwan, especially for recipients of hematopoietic stem cells from mismatched unrelated and haploidentical donors. Preemptive ganciclovir therapy should be initiated when the CMV viral load exceeds 1000 copies/mL, and should not be discontinued until CMV DNA is no longer detected in the blood. For allo-HSCT recipients who have CMV-related diseases, ganciclovir with or without CMV-specific intravenous immunoglobulin is the standard of care. The limited availability of foscarnet, an alternative for patients who are not responsive to or cannot tolerate ganciclovir, is a crucial issue in Taiwan. For pediatric allo-HSCT recipients, more data are needed to propose a CMV management recommendation.     

不同预处理移植后树突状细胞早期重建及与移植物中CD34+细胞相关性分析

目的：比较不同预处理异基因造血干细胞移植（allo-HSCT）后早期树突状细胞（DCs）亚群重建情况，以及移植物中CD34^＋细胞是否影响移植后早期DCs亚群重建。方法：采用三色流式细胞仪动态检测不同预处理移植后早期外周血树突状细胞亚群DC1、DC2水平。结果：移植后早期清髓性移植患者体内DCs亚群数量非常低，常规移植组移植后14天与半相合移植组相比，DC1、DC2均无统计学意义（P〉0．05）。非清髓性移植组（NST）DC1、DC2高于清髓性移植组，两者相比具有统计学意义（P〈0．05）。在30天和60天，所有组DC1、DC2略有波动，但是幅度不大。以输入的CD34^＋细胞数平均分为三组，三组患者DC1、DC2移植后14、30和60天均无统计学意义（P〉0．05）。结论：NST后患者早期DCs重建较清髓性干细胞移植患者早，而常规移植和半相合移植早期DCs重建较慢，二者无差别。移植物中的CD34^＋细胞不影响移植后早期DCs亚群重建。     

Bone marrow-derived mesenchymal stem cells decrease acute graft-versus-host disease after allogeneic hematopoietic stem cells transplantation

Graft-versus-host disease is a major complication of allogeneic hematopoietic stem cells transplantation, leading to serious morbidity and mortality. Mesenchymal stem cells(MSC)from bone marrow cause immunoregulation in vitro and in vivo. They also have the potential to protect from lethal GVHD after both autologous and allogeneic Hematopoietic Stem Cells Transplantation (HSCT). In this study, we investigated the mechanisms responsible for GVHD in the allo-HSCT co-transplantation with MSC condition. The model of acute GVHD in Rats was established using allogeneic HSC with donor-derived T cells transplantation, with or without additional donor-derived MSC co-transplantation. The degrees of GVHD were compared, the differentiation of CD4⁺, CD8⁺, Th1/Th2 and CD4⁺CD25⁺ T cells in vivo were assessed by flow cytometry and RT-PCR analyses. We found that MSC inhibited lethal GVHD after allo-HSCT. The value of CD8⁺ and CD4⁺ T cells and the ratio of Th1/Th2 T cell subsets decreased, at the same time the proportion of CD4⁺CD25⁺ T cells increased both in spleen lymphocytes and thymocytes in vivo after allo-HSCT with MSC co-transplantation compared with conventional allo-HSCT. Our results strongly suggested that BM-derived MSC has the function of preventing lethal GVHD after allo-HSCT by means of homeostasis of T subsets in vivo.     

异基因造血干细胞移植CD94分子在T细胞高表达的意义初探

目的检测CD94分子在T细胞表达的水平,同时探讨在异基因造血干细胞移植(HSCT)中发生移植物抗宿主病(GVHD)患者CD94分子在T细胞表达的意义。方法HSCT治疗高危白血病和遗传性溶血性贫血成功植入的儿童患者,其中同胞脐血移植(UCBT)10例,非血缘相关UCBT1例,异基因外周血造血干细胞移植(alloPBSCT)5例,在移植前后发生GVHD时采用流式细胞仪检测和比较外周血中CD94的表达,并与正常外周血水平比较。结果CD94主要表达于CD3+CD8+T细胞,正常情况下外周血和脐血T细胞表达率低于10%,其中CD4+细胞几乎不表达,低于2%。但在UCBT或alloPBSCT后CD94在CD4+T细胞和CD8+T细胞均明显增高。3例UCBT无GVHD,其余均发生了ⅠⅣ0急性GVHD。急性GVHD发生时CD4+CD94+和CD8+CD94+T细胞表达明显升高。结论异基因造血干细胞移植后发生GVHD时,T细胞高表达CD94,可能是在同种抗原的刺激下机体自我保护的结果。     

异基因造血干细胞移植后的相关并发症及危险因素

背景：有效预防和治疗异基因造血干细胞移植后并发症是提高患者存活率的重要因素。 目的：分析异基因造血干细胞移植后相关并发症的发生和危险因素。 方法：应用文献检索的方法获取异基因造血干细胞移植后相关并发症研究的文献,对符合研究标准的文献进行深入的数据分析,文章选取异基因造血干细胞移植后极易发生的并发症进行分析,如肺部并发症、真菌性败血症、巨细胞病毒感染以及中枢神经系统并发症等。 结果与结论：异基因造血干细胞移植后易出现肺部并发症,而且死亡率较高,肺部并发症的发病机制可能与移植物抗宿主病和巨细胞病毒抗原血症相关。异基因造血干细胞移植后真菌性败血症病原菌以假丝酵母菌属为主,死亡率较高,应二级预防性和早期经验性抗真菌治疗。更昔洛韦、膦甲酸钠对异基因造血干细胞移植后巨细胞病毒感染的治疗有效。中枢神经系统并发症在异基因造血干细胞移植后发生率较低,但在治疗过程也不容忽视。异基因造血干细胞移植后相关并发症的发生与多种危险因素有关,在临床治疗过程中要对相关因素采取预防措施,减少并发症的发生,提高患者的存活率。     

Assessment of lineage-specific chimerism after allogeneic stem cell transplantation

IntroductionDonor lineage-specific chimerism of hematopoietic cells enables very precise monitoring of engraftment in selected cell lines after allogeneic stem cell transplantation (allo-SCT).Materials and methodsThe study group consisted of 12 acute leukemia patients who underwent allo-SCT in the Department of Hematology and Bone Marrow Transplantation in Katowice, Poland. Lineage-specific chimerism was assessed in B cells (CD19+ CD38−/+), plasma cells (CD19+ CD38++), T cells (CD3+ or CD7+ CD56−), monocytes (CD14+), and immature progenitor cells deriving from myeloid line (CD34+CD19). We also assessed erythrocyte chimerism by flow cytometry.ResultsAll patients engrafted. 8 out of 10 patients presented normal donor hematopoiesis. Lineage specific chimerism in these patients corresponded with chimerism analysis in unsorted material and with undetectable minimal residual disease (MRD). Relapse of the underlying disease was diagnosed in 2 patients. In both cases loss of donor chimerism occurred in leukemia specific cell line and corresponded with detectable MRD. One patient with secondary graft failure presented decreasing lineage specific chimerism in all subpopulations, with negative MRD status. In 10 patients normal hematopoiesis of donor-origin was assessed by flow cytometry. In one case no donor-derived erythrocytes were detected and the diagnosis of pure red cell aplasia was set.ConclusionsLineage specific chimerism as a method of high sensitivity and specificity allows for precise assessment of donor chimerism especially in clinically ambiguous situations. Assessment of erythrocyte chimerism by flow cytometry is a reliable method of monitoring erythroblastic line engraftment. Presented results are preliminary and the study is being continued.     

骨髓间质干细胞降低大鼠移植物抗宿主反应的实验研究

目的 探讨MSCs对GVHD的作用及其机制.方法 建立大鼠同种异体骨髓移植模型,同时输入供者的T淋巴细胞诱导出移植物抗宿主反应,联合或不联合移植供体来源的MSCs,观察受鼠的生存时间,同时利用RT-PCR法研究Th1/Th2淋巴细胞亚群的比例,用ELISA法检测移植后体内IL-4细胞因子的浓度.结果 GVHD组的平均生存时间为(17.30±2.33)天,实验组的平均生存时间为(24.10±2.36)天 , 与单独移植HSCs相比,MSCs与HSCs共移植明显延长的受鼠的生存时间.同时,GVHD组Th1/Th 2 细胞比值为1.29±0.06,IL-4因子的浓度平均为(14.84±2.59) pg/mL,实验组Th1/Th 2细胞比值为(0.77±0.14),IL-4因子的浓度平均为(40.09±13.99) pg/mL.MSCs与 HSCs 共移植降低了体内Th1/Th2淋巴细胞亚群的比例,提高了体内IL-4细胞因子的浓度.结论 MSCs与HSCs共移植能有效抑制HSCs移植后致死性GVHD的发生,延长生存时间,同时MSCs 可能通过作用于体内Th1/Th2淋巴细胞亚群的比例,促进体内IL-4细胞因子的分泌从而间接发挥了抑制GVHD的作用.     

Comparison of six different specimen types for Epstein-Barr viral load quantification in peripheral blood of pediatric patients after heart transplantation or after allogeneic hematopoietic stem cell transplantation

Background

Epstein-Barr Virus (EBV) a gamma-herpes virus is associated with a spectrum of lymphoid and epithelial malignancies including posttransplant lymphoproliferative disorders (PTLD). EBV-load measurement has been shown to be important for the monitoring of these patients. However, in contrast to the viral quantification of human immunodeficiency virus or human hepatitis C virus, the EBV-load measurement has not been completely standardized as yet.

Objectives

In this study, we compared the EBV DNA levels in whole blood (WB), plasma, peripheral mononuclear cells (PBMC) and B-cells (BC) in children and adolescents after heart transplantations (HTx) and allogeneic hematopoietic stem cell transplantations (HSCT).

Study design

In a period of 2 years (from May 2007 to May 2009) we collected 547 samples of 96 cardiac transplant recipients and 248 samples of 37 patients who underwent HSCT. For EBV DNA quantification we used a duplex real-time PCR (ABI Prism 7500, Applied Biosystems). Additionally, EBV-load of PBMC and BC were normalized with respect to endogenous cell DNA.

Results

In both patient populations we found no significant difference of test sensitivity for the EBV detection. In PBMC as well as BC, there was a high correlation between the analysis of cells with and without normalization in both populations. Spearman's correlation coefficient ρ between PBMC without and PBMC with normalization was ρ = 0.98 (P < 0.0001) in patients after HTx and ρ = 0.99 (P < 0.0001) in patients after HSCT. Correlation between BC with and without normalization was ρ = 0.98 (P < 0.0001) in patients after HTx and ρ = 0.995 (P < 0.0001) in patients after HSCT. When comparing the different blood compartments for EBV quantification in both populations, the strongest correlations were found between the EBV DNA levels in WB and PBMC (HTx: ρ = 0.93, P < 0.0001; HSCT: ρ = 0.81, P < 0.0001) followed by PBMC and BC (HTx: ρ = 0.87, P < 0.0001; HSCT: ρ = 0.81, P < 0.0001) as well as WB and BC (HTx: ρ = 0.86, P < 0.0001; HSCT: ρ = 0.75, P < 0.0001). In contrast, the correlation coefficients between plasma and the other blood compartments (WB as well as PBMC or BC) were lower.Six patients developed seven episodes of PTLD (five patients after HTx and one after renal transplantation). Analyzing the different blood compartments, we found that a threshold of WB ≥ 20,000 EBV-copies/ml and plasma ≥ 1000 EBV-copies/ml had the highest sensitivities and specificities (WB: sensitivity 100%, specificity 87% and plasma: sensitivity 88%, specificity 98%).

Conclusion

Normalization towards an endogenous control does not seem to be necessary for EBV quantification in peripheral blood. The analysis of whole blood correlates well with B-cells and PBMC. Routine screening of EBV DNA in whole blood appeared to be a useful tool supplemented by EBV-load measurement in plasma to discriminate chronic high EBV-load carrier without risk for PTLD from those who are at risk for PTLD. Values in whole blood higher than 20,000 EBV-copies/ml WB and plasma values higher than 1000 EBV-copies/ml plasma indicated PTLD in our series.     

Adoptive precursor cell therapy to enhance immune reconstitution after hematopoietic stem cell transplantation

Strategies to enhance post-transplant immune reconstitution without aggravating graft-vs-host disease (GVHD) can improve the outcome of allogeneic hematopoietic stem cell transplantation. Recent preclinical studies demonstrated that the use of T cell depleted allografts supplemented with committed progenitor cells (vs stem cells only) allows enhanced immune reconstitution of specific hematopoietic lineages including myeloid, B, T, and natural killer lineages in the absence of GVHD. This novel adoptive therapy resulted in significantly improved resistance to microbial pathogens and could, in some cases, even mediate tumor immunity. Clinical protocols using adoptive transfer of committed hematopoietic progenitor cells are currently being evaluated.