异基因造血干细胞移植后的长期随访*

背景：近20年来中国的造血干细胞移植工作迅速发展,接受异基因造血干细胞移植后得到治愈的患者越来越多,但关于患者移植后长期随访情况的报道少见。 目的：总结异基因造血干细胞移植后患者的长期随访情况。 方法：对1995/2005在中山大学附属第三医院进行异基因造血干细胞移植的30例患者进行长期随访,总结患者的远期并发症、长期生存率、死亡原因,调查长期生存患者的生活质量。 结果与结论：30例患者的慢性移植物抗宿主病发生率为67%,所有患者均出现不育,1例轻度肺纤维化,无其他远期并发症;移植后2年的无病生存率为60%,2年内未复发的患者均长期生存,生存时间最长的1例已超过14年;12例患者死亡,均发生于移植后2年内,其中5例死于复发,7例为移植相关死亡(均死于感染);长期生存的患者移植后1年时的生活质量较差,随着生存时间的延长生活质量逐渐改善,5年时的生活质量明显提高,8年时多已拥有较高的生活质量,影响生活质量的主要因素是慢性移植物抗宿主病。     

异基因造血干细胞移植中急性移植物抗宿主病的研究状况

目前认为,异基因造血干细胞移植可通过免疫细胞介导的移植物抗白血病效应消除体内残留的肿瘤细胞,达到治愈的目的。但移植物抗宿主病,尤其急性移植物抗宿主病,是异基因造血干细胞移植应用中最棘手的问题,是移植后致残及相关死亡的主要原因。而控制移植物抗宿主病的药物大多抑制移植物抗白血病效应,同时付出了移植效能降低、感染机会增加的代价。因此,理想的方法是寻找使T细胞处于激活状态,保持移植物抗白血病效应,但同时又抑制特异性同种抗原。一些对经典抗原提呈细胞、树突状细胞等的新发现,移植方式的改良以及间充质干细胞和调节性T细胞的应用等,为移植物抗宿主病的防治提示了新的方向。     

非清髓性异基因造血干细胞移植基础及临床研究进展

非清髓性异基因造血干细胞移植现在被广泛的用在由于年龄或合并症而不适应行传统HCT的患者身上,采用非清髓性预处理策略,移植后形成供受体造血细胞混合嵌合状态,淋巴细胞供受体双向免疫耐受,即非清髓性干细胞移植。通过非清髓性预处理方案,移植后形成供受体细胞嵌合状态,发挥移植物抗肿瘤效应,来达到治疗目的,与传统HCT中文？相比有较低的移植相关死亡率,术后GVHD中文？发生减少,且扩大了适应症范围,并且非清髓性预处理方案的改进、移植后相关疾病发生的防治可以进一步提高治疗效果。虽然有许多治疗策略被提出,并在动物模型上取得了成功,但其在人体方面的应用仍需得到进一步的证实和研究。     

骨髓间充质干细胞移植对心肌梗死后炎性细胞因子表达的调节*☆

目的：近来研究表明，骨髓间充质干细胞具有独特的免疫调节特性，实验拟观察骨髓间充质干细胞移植对梗死心脏炎性细胞因子表达的影响，以进一步认识干细胞治疗心肌梗死的可能机制。 方法：实验于2006-11/2007-08在中南大学湘雅二医院心内科实验室和实验动物中心完成。①实验材料：清洁级SD大鼠由中南大学湘雅二医院实验动物中心提供，实验过程中对动物处置符合动物伦理学标准。②实验方法：提取体质量120~150 g SD大鼠骨髓间充质干细胞，以溴脱氧尿嘧啶核苷标记。取体质量180~200 g 雄性SD大鼠，结扎冠状动脉前降支建立心肌梗死模型，将模型随机分为3组：假手术组：仅穿线不接扎血管；PBS溶液注射组：模型制作成功即刻心外膜注射PBS液；干细胞移植组：模型制作成功即刻心外膜注射溴脱氧尿嘧啶核苷标记的骨髓间充质干细胞，每组8只。③实验评估：于术后1 ，28 d行心脏超声检测，测量大鼠射血分数、左室短轴缩短率和左室舒张末期直径以评价心脏功能，并进行免疫组织化学和western blot检测，分析移植细胞存活和炎性细胞因子的表达情况。 结果：24只建模成功大鼠均进入结果分析。①术后28 d，心脏超声检测显示与注射PBS溶液相比，移植骨髓间充质干细胞可以延缓心室重构，改善心功能[射血分数：（30.76±3.39）%，（24.06±4.71）%，P < 0.05；左室短轴缩短率：（29.33±4.87）%，（23.05±3.94）%，P < 0.05；左室舒张末期直径：（6.45±0.47）mm，（7.81±0.31）mm，P < 0.05] 。②免疫组织化学检测显示，心肌梗死后即刻心外膜移植的骨髓间充质干细胞可以在宿主体内存活。③ 与注射PBS溶液相比，骨髓间充质干细胞移植可以明显降低肿瘤坏死因子α、白细胞介素6表达（P < 0.05） ，同时增加白细胞介素10表达（P < 0.05) 。western blot检测与免疫组织化学结果一致。 结论：骨髓间充质干细胞移植可以延缓心肌梗死大鼠心室重构，同时下调大鼠梗死心脏促炎细胞因子肿瘤坏死因子α和白细胞介素6表达，上调抗炎细胞因子白细胞介素10表达。     

自体分选造血干细胞和异体脐带间充质干细胞移植治疗神经系统变性疾病

背景：将自体分选造血干细胞和异体脐带间充质干细胞技术应用于临床,试图寻找一种新的治疗神经系统变性疾病的可行方法。 目的：探讨自体分选CD34+造血干细胞和异体脐带间充质干细胞治疗神经系统变性病的可行性。 方法：入选神经系统变性病患者21例,其中运动神经元病15例,脊髓小脑共济失调6例。取4 mL的脐带间充质干细胞或自体分选干细胞液经腰穿注射到患者蛛网膜下腔,每次注射的细胞数1.0×107。于干细胞治疗后3个月进行评分。 结果与结论：15例运动神经元病患者,治疗总有效率为80%,治疗前后的脊髓侧索硬化功能分级量表和自我评估问卷评分差异有显著性意义(P < 0.05);6例脊髓小脑共济失调患者,按ICARS评分,4例病情等级下降,治疗总有效率为77%。21例中4例有轻度低颅压性头痛(腰穿后),14例采用脐带间充质干细胞治疗的患者中2例在治疗后2 h出现短暂发热,其余患者治疗后未见明显不良反应。结果初步表明自体分选造血干细胞和异体脐带间充质干细胞治疗神经系统变性病临床疗效肯定,安全可行。     

单倍体相合造血干细胞移植后慢性移植物抗宿主病小鼠模型的 建立及其评价*★

目的：因供受者主要或次要组织相容性抗原存在差异而引起的移植物抗宿主病是影响异基因造血干细胞移植效果的关键因素，目前国内尚缺乏研究此领域的实验动物模型及其评价体系。本实验拟建立半相合异基因造血干细胞移植后慢性移植物抗宿主病小鼠模型，并制定半定量评价体系。 方法：实验于2007-03/08在南方医科大学实验动物中心完成。①动物：供鼠为近交系Balb/CH-2d小鼠40只，雄性，8~10周龄，体质量18~22 g；受鼠为CB6F1小鼠(Balb/c×C57BL/6 F1H-2d/b) 48只，雌性，10~12周龄，体质量18~24 g，随机数字表法分为3×107，6×107，9×107个脾细胞移植组及空白对照组，12只/组。小鼠均由南方医科大学实验动物中心提供，SPF级，实验过程中对动物的处置符合动物伦理学标准。②实验方法：Balb/CH-2d小鼠颈椎脱臼处死，取脾脏研磨后制成脾单细胞悬液，离心弃上清，加入Tris-NH4Cl裂解红细胞，用RPMI 1640调整细胞浓度为1.2×108 L-1，锥虫蓝染色计数脾细胞拒染率为95.6%。3×107，6×107，9×107个脾细胞移植组分别经尾静脉输入对应数量的MHC半相合脾细胞悬液0.5 mL，空白对照组输入等体积 RPMI 1640培养液。③实验评估：脾细胞移植后2，5，8，12周，光镜下计数20个分裂相，检查骨髓细胞中的Y染色体数量，进行嵌合体分析。脾细胞移植18 d后，每3 d观察受体小鼠的临床表现，包括体质量、体形、体位、毛发变化及生存情况，并予以评分。输注脾细胞后100 d观察皮肤、肝脏、回盲端小肠靶器官的病理变化，并进行评分。 结果：48只受体小鼠均进入结果分析。①嵌合体检测：6×107个脾细胞移植组可形成较为稳定的低水平供受者混合嵌合体；9×107个脾细胞移植组起初可形成较高水平的供受者混合嵌合体，随时间的推移水平降低；3×107个脾细胞移植组未能形成稳定的供受者混合嵌合体。②慢性移植物抗宿主病的临床及病理评分：移植物抗宿主病发生时间多集中在供鼠脾细胞输注后30~80 d，临床表现以皮肤改变、体质量减轻、弓背体态为主，病理损害以皮肤、肠道及肝脏明显。与3×107个脾细胞移植组比较，6×107，9×107个脾细胞移植组临床和病理评分均显著升高（P < 0.05），慢性移植物抗宿主病发生率均显著升高（P < 0.01）；后2组临床和病理各项指标之间差异均无显著性意义（P > 0.05）。 结论：输注6×107和9×107个脾细胞的小鼠成功诱导出半相合异基因造血干细胞移植慢性移植物抗宿主病，其临床和病理评价体系具有较高的一致性和实用性，为进一步分析慢性移植物抗宿主病的发病机制和评价干预因素的优劣奠定实验学基础。     

异基因造血干细胞移植后慢性粒细胞白血病患者bcr-abl融合基因水平的动态变化

背景：bcr-abl融合基因不仅是慢性粒细胞白血病的诊断标准，其水平的变化也是判断病情或疗效的惟一指标，尤其是bcr-abl融合基因的动态变化对于异基因造血干细胞移植效果的评估和预后判断可能更为重要。 目的：了解慢性粒细胞白血病患者异基因造血干细胞移植后bcr-abl融合基因水平的变化情况。 设计：回顾性病例分析。 对象：2001-04/2007-01在广东省人民医院血液科收治的确诊1年内行异基因造血干细胞移植的慢性粒细胞白血病患者22例，中位年龄31.5岁，患者对治疗均签署知情同意书，治疗方案经医院医学伦理委员会批准。 方法：所有同胞供者均采用粒细胞集落刺激因子行外周血干细胞动员。所有患者均采用改良BU/CY联合FLU预处理方案，供受者血型不相合者于移植前7 d、1 d行血浆置换以降低血型抗体滴度。患者经深静脉输入同胞供者平均单个核细胞数为5.48×108/kg，平均CD34+细胞数为10.45×106/kg。由于移植和随访时间不同，每例患者收集标本3~14份，共130份外周血标本。 主要观察指标：通过实时荧光定量PCR法检测22例患者异基因造血干细胞移植后不同时间130份外周血标本中bcr-abl融合基因的水平。 结果：移植后6个月内，bcr-abl融合基因总体水平逐渐降低，移植后1、3、6个月bcr-abl阴性患者的比例分别为33.33%、53.33%和84.62%，bcr-abl高水平阳性（bcr-abl/abl ≥ 0.02%）患者的比例分别为58.33%、33.33%和15.38%，低水平阳性（bcr-abl/abl < 0.02%）患者的比例分别为8.33%、13.33%和0。移植6个月后，多数患者连续多次检测不到bcr-abl融合基因，其中持续阴性者9例（69.23%），低水平波动者1例（7.69%），低水平持续阳性者1例（7.69%），复发2例（15.38%）；复发的2例患者移植后6个月内连续检测bcr-abl融合基因水平均未降低至0，且3次检测结果中2次属于高水平阳性。 结论：异基因造血干细胞移植后，慢性粒细胞白血病患者bcr-abl融合基因总体水平逐渐下降，但不同患者bcr-abl变化情况各异，通过实时定量PCR检测可明确其变化趋势。     

Clinical and MRI outcome after autologous hematopoietic stem cell transplantation in MS

The authors report the outcome of 14 patients with severe multiple sclerosis treated with autologous hematopoietic stem cell transplantation (AHSCT) after a median follow-up period of 3 years. The 3-year actuarial probability of progression-free survival was 85.7% and that of disease activity-free survival was 46.4%. On MRI, no T1-enhanced lesions were detected after AHSCT. The mean change in T2 lesion volume from baseline to the third year was -20.2% and that of the corpus callosum area was -12.7%; 50% of this reduction was seen during the first year.     

Autologous and allogeneic hematopoietic stem cell transplantation for Multiple Sclerosis: perspective on mechanisms of action

Multiple Sclerosis (MS) is a frequent demyelinating immune-mediated disease of the central nervous system (CNS) that affects principally young adults and leads to severe physical and cognitive impairment. The current standard treatment makes use of the immune modulators beta-interferon, glatiramer acetate and natalizumab, or immunosuppressants such as mitoxantrone. However, these agents are only partially effective and in a number of patients fail to achieve satisfactory disease control. Autologous hematopoietic stem cell transplantation (HSCT) is being explored in the treatment of severe MS as a means of delivering high-dose immunosuppression followed by 'rescue' of the immuno-hematopoietic system with autologous HSC. The potential therapeutic benefit is based on the concept of so-called 'resetting' the immune system. The use of allogeneic HSCT as a possible therapeutic approach for severe MS is inspired by case reports of MS patients that underwent allogeneic HSCT for a concomitant hematological malignancy, and subsequently is supported by data from rodent models of MS. Allogeneic HSCT may offer specific therapeutic effects, such as the replacement of the autoreactive immune compartment by healthy allogeneic cells and the development of a graft-versus-autoimmunity (GVA) effect. Here, we review the currently available experimental and clinical evidence to support the role of autologous and allogeneic HSCT in MS.     

HLA相合同胞异基因外周血造血干细胞移植治疗急性白血病

背景：HLA相合同胞间异基因外周血造血干细胞移植是治疗急性白血病的一种有效方法。 目的：评价HLA相合异基因外周血造血干细胞移植治疗急性白血病的临床疗效及并发症。 方法：25例急性白血病患者接受HLA相合同胞的异基因外周血造血干细胞移植,其中急性髓系白血病20例,急性淋巴细胞白血病5例。预处理方案为BU+CY方案或CY+TBI方案,移植物抗宿主病预防采用环孢素A+吗替麦考酚酯+短程甲氨蝶呤。 结果：最短随访2个月,最长随访80个月。患者均获造血重建,中性粒细胞≥0.5×109 L-1的时间为10~18 d,血小板≥20× 109 L-1的时间为10~37 d。主要并发症：感染败血症12例,巨细胞病毒感染9例,带状疱疹病毒感染3例,发生急性移植物抗宿主病10例,慢性移植物抗宿主病11例,出血性膀胱炎4例。至随访结束,17例无病生存,8例死亡。提示HLA相合同胞异基因外周血造血干细胞移植是治疗急性白血病安全有效的方法。     

Etiology,characteristics and outcome of seizures after pediatric hematopoietic stem cell transplantation

PurposeEpileptic seizures are frequent manifestations after hematopoietic stem cell transplantation (HSCT). In this retrospective single-center study we evaluated electroclinical features and analyzed etiologies and outcome of seizures after pediatric HSCT.MethodsOf 261 children transplanted between 2000 and 2010, we identified and analyzed data of 28 patients with seizures within a year from HSCT.ResultsMost frequent etiologies were posterior reversible encephalopathy syndrome (PRES, 14 patients) and central nervous system (CNS) infections (4 patients). Seizures were the presentation of the underlying complications in 22 patients. Sixteen episodes of status epilepticus were identified. Seizures secondary to PRES were usually longer and associated with non-convulsive signs. Early neuroimaging and EEG monitoring proved to be crucial to diagnose and treat seizures and their causes. No patients developed epilepsy suggesting that chronic antiepileptic therapy is not necessary in these patients. Overall survival was 32.3% over 5 years in patients with seizures and 45.8% in patients without seizures (p < 0.05). Multivariate statistical analysis identified as independent risk factors for seizures a diagnosis of non-oncological disease and cord blood stem cell transplantation.ConclusionsSeizures in transplanted children are a severe event and are associated with high morbidity and poor outcome. In particular, patients with non-oncological diseases and cord blood stem cell transplantation have to be considered at high risk of seizures. Moreover, this study underlines the importance of early recognition of non-convulsive clinical signs and of EEG monitoring for a prompt diagnosis and an appropriate management of seizures and their causes.     

MRI and CSF oligoclonal bands after autologous hematopoietic stem cell transplantation in MS

OBJECTIVE: To analyze the MRI and CSF oligoclonal bands (OB) changes in patients with MS who underwent an autologous hematopoietic stem cell transplantation (AHSCT). BACKGROUND: AHSCT is evaluated as an alternative therapy in severe MS. In previous series of AHSCT for MS, data on MRI or OB outcome were limited or not provided. METHODS: Five patients with a median Kurtzke's EDSS score of 6.5, more than two attacks, and confirmed worsening of the EDSS in the previous year received an AHSCT. Hematopoietic stem cells were mobilized with cyclophosphamide (3 g/m2) and granulocyte colony-stimulating factor (5 microg/kg/d). The graft was T cell depleted by positive CD 34+ selection. Conditioning regimen included BCNU (300 mg/m(2)), cyclophosphamide (150 mg/kg in 3 days), and antithymocyte globulin (60 mg/kg in 4 days). MRI scans were scheduled at baseline and 1, 3, 6, and 12 months and OB analysis at baseline and 3 and 12 months post-AHSCT. RESULTS: Four patients had a stable or improved EDSS after a median follow-up of 18 months (range, 12 to 24 months). The fifth patient's condition deteriorated during AHSCT. She partially improved and remained stable after month 3 after AHSCT. The baseline CSF OB persisted 1 year after AHSCT. MRI studies after AHSCT showed no enhanced T1 lesions and no new or enlarging T2 lesions. The median percentage change of T2 lesion load was -11.8% (range, -26.6 to -4.0%). All patients had a decrease of corpus callosum area at 1 year (median, 12.4%; range, 7.8% to 20.5%) that did not progress in the two patients evaluated at 2 years after AHSCT. CONCLUSIONS: Although the persistence of CSF OB suggests the lymphocytes were not eliminated from the CNS, the follow-up MRI studies showed no enhanced T1 brain lesions and a reduction in the T2 lesion load that correlated with the clinical stabilization of MS after AHSCT.     

含氟达拉滨预处理方案对异基因造血干细胞移植过程中造血重建、移植物抗宿主病及患者生存的影响

目的：观察接受不同预处理方案改良Bucy(白消安/阿糖胞苷/环磷酰胺)或白消安/氟达拉滨对异基因造血干细胞移植后造血重建,移植物抗宿主病及生存的影响。 方法：选择2006-12/2008-06解放军北京军区总医院收治的行异基因造血干细胞移植恶性血液病患者27例,按移植预处理方案不同分为两组,10例给予以氟达拉滨+白消安为基础的预处理方案：移植前2~6 d氟达拉滨 30 mg/(m2•d),移植前3~6 d白消安 3.2 mg/(kg•d);17例给予以改良Bucy为基础的预处理方案：移植前4~6 d白消安4 mg/(kg•d),移植前两三天环磷酰胺60 mg/(kg•d)和阿糖胞苷2.0 g/(m2•d)。比较两组患者移植后白细胞、血小板重建时间,移植物抗宿主病的发生及其程度,复发与移植相关死亡率。 结果：白细胞、血小板重建时间在Bucy组与白消安/氟达拉滨组分别为[(8.45±2.31),(8.96±2.47)d,P=0.957;(13.31±4.80),(15.89±5.21) d,P=0.662]。Bucy组与白消安/氟达拉滨组患者急性移植物抗宿主病发生率分别为[47%(8/17),40%(4/10),P=0.629]。27例患者截至统计时间10例死亡,其中复发死亡4例,移植物抗宿主病死亡3例,感染死亡2例,其他移植合并症死亡1例。Bucy组与白消安/氟达拉滨组1年内总生存率分别为(58.30±19.80)%和(73.00±11.80)%,1年内无病生存率为(48.60±18.70)%和(72.20±12.80)%,白消安/氟达拉滨组总生存率及无病生存率均较Bucy组高,但两组1年内总生存率、无病生存率差异均无显著性意义(P=0.511,0.854)。 结论：氟达拉滨应用于移植预处理方案中,可降低异基因造血干细胞移植过程中髓外毒性,但对疗效并无明显影响。     

恶性血液病患者接受造血干细胞移植后发生糖尿病的危险因素分析★

目的：造血干细胞移植是目前根治血液病的惟一方法。调查分析恶性血液病造血干细胞移植后糖尿病发生及发展的危险因素。 方法：①对象和分组：收集2003-01/2007-02武汉市中心医院行造血干细胞移植后患者和外院行造血干细胞移植后转至武汉市中心医院的患者16例。根据干细胞移植术后血糖水平分为糖尿病组6例，非糖尿病组11例，两组平均年龄45.2岁。②评估：分析移植后糖尿病的发生与患者年龄、术前疾病状态（化疗次数、大剂量强化的次数、血压状态、糖耐量情况）、免疫抑制剂、皮质激素用量及时间的关系，以及移植后糖尿病对患者的影响。 结果：16例患者均进入结果分析。①术后出现5例糖尿病；术前有糖耐量异常者其移植后糖尿病的发生率高于无糖耐量异常者。②年龄大于40岁、曾经多次使用过含有糖皮质激素化疗方案的患者，其糖尿病的发病率较高；激素半年内撤离的患者移植后糖尿病的发生率明显低于半年内未撤离者。③两组患者使用免疫抑制剂及血药浓度的情况对糖尿病的发生无影响。④糖尿病组对术后巨细胞病毒感染率无明显影响。 结论：年龄大于40岁、曾多次使用过含糖质皮激素的化疗方案、糖耐量异常、长期使用糖皮质激素预防移植物抗宿主反应者是糖尿病的高危人群。     

丁基苯酞加强造血干细胞移植及内源性干细胞动员治疗脑梗死的效应

Exogenous stem cell transplantation and endogenous stem cell mobilization are both effective for the treatment of acute cerebral infarction. The compound dl-3-butylphthalide is known to improve microcirculation and help brain cells at the infarct loci. This experiment aimed to investigate the effects of dl-3-butylphthalide intervention based on the transplantation of hematopoietic stem cells and mobilization of endogenous stem cells in a rat model of cerebral infarction, following middle cerebral artery occlusion. Results showed that neurological function was greatly improved and infarct volume was reduced in rats with cerebral infarction. Data also showed that dl-3-butylphthalide can promote hematopoietic stem cells to transform into vascular endothelial cells and neuronal-like cells, and also enhance the therapeutic effect on cerebral infarction by hematopoietic stem cell transplantation and endogenous stem cell mobilization.     

Quantitative proton magnetic resonance spectroscopy of children with adrenoleukodystrophy before and after hematopoietic stem cell transplantation

About 35-40 % of boys with X-linked adrenoleukodystrophy (ALD) develop a rapidly progressive cerebral form which leads to severe neurologic disability and death within 3-5 years after onset of clinical symptoms. Because previous proton magnetic resonance spectroscopy (MRS) studies of ALD identified metabolite patterns characteristic of demyelination, gliosis, and neuroaxonal loss, this work tested the hypothesis that MRS--apart from indicating disease progression--provides criteria for the outcome after hematopoietic stem cell transplantation (HSCT) which has been promising at an early stage of the active disease. Follow-up quantitative proton MRS was performed in frontal and occipital white matter of ALD patients (n = 12) before and up to 5 years after HSCT. The observed metabolite alterations were retrospectively correlated with the clinical outcome representing either a stable condition (n = 5), a further deterioration (n = 5), or death (n = 2). While disease progression of patients before HSCT was mainly characterized by a further increase of elevated choline-containing compounds (Cho) as an indicator of active demyelination, a positive outcome after HSCT was correlated with high N-acetylaspartate (tNAA) levels in affected white matter before HSCT yielding positive and negative predictive values for tNAA of 80 %. Although to be confirmed in a larger cohort of patients, the present findings suggest the preservation of neuroaxonal integrity as a prerequisite for an arrested course. Conversely, the combination of increased Cho with markedly reduced tNAA before HSCT apparently reflects a degree of tissue degeneration which precludes a successful therapeutic intervention.     

Discordant functional and inflammatory parameters in multiple sclerosis patients after autologous haematopoietic stem cell transplantation

This article describes outcomes in four patients with advanced multiple sclerosis up to two years after autologous haematopoietic stem cell transplantation using a total-body irradiation-based preparative regimen. MRI and CSF analyses demonstrated clear suppression of the inflammatory processes. The results demonstrate however, a dissociation of inflammation parameters and functional disability findings raising questions about optimal future stem cell transplantation strategies for this disease.