混合造血干细胞移植后供者淋巴细胞输注加 IL-2治疗急性非淋巴细胞白血病

为了发挥异基因造血干细胞移植(Allo—SCT)白血病复发率低和自体造血干细胞移植并发症少、应用范围广的优点我们首先开展了混合HLA半相合异体骨髓的自体骨髓移植，实验和临床研究证明混合异体骨髓的自体骨髓移植能部分发挥异体骨髓移植产生的移植物抗白血病(GVL)作用，减少移植后白血病的复发，并且为复发患者进行供者淋巴细胞输注(DLI)提供了契机。本文首次报告混合移植后DLI加IL-2免疫治疗急性白血病2例如下。     

单倍型相合骨髓移植治疗伴髓外侵犯的难治复发变异型急性早幼粒细胞白血病获长期生存1例并文献复习

目的:探讨单倍型相合骨髓移植治疗伴有髓外侵犯的难治性变异型急性早幼粒细胞白血病的疗效。方法:采用母亲供髓的单倍型相合移植治疗1例伴髓外侵犯的难治复发变异型急性早幼粒细胞白血病,预处理方案为CyTBI加Ara-c,GVHD预防采用联合免疫抑制剂和供者应用G-CSF的方法,联合免疫抑制剂包括环胞素A、短程氨甲碟呤、霉酚酸酯、CD 25单抗和抗胸腺细胞球蛋白的应用。结果:移植后+1个月造血重建,植入成功,免疫功能逐渐恢复,定期随访,PML/RARα融合基因持续(-),现已无病生存76个月。结论:单倍型相合骨髓是治愈高危难治复发急性早幼粒白血病的有效措施,为患者提供了长期生存的机会。     

HLA半相合血缘性骨髓移植治疗慢性粒细胞白血病4例

目的:探索半相合未去除T细胞骨髓移植治疗慢性粒细胞白血病的可行性。方法:4例慢性粒细胞白血病患者接受HLA1或2个位点不相合亲缘骨髓移植。用阿糖胞苷、环磷酰胺和全身照射进行预处理,供者应用GCSF250μg/d,连用7d后采髓。移植物抗宿主病(GVHD)预防除用环孢菌素A(CsA)和甲氨蝶呤(MTX)外,在移植前第4天～第1天用抗胸腺细胞球蛋白ATG(兔抗)2.5mg/(kg·d),移植后第7天开始加用霉酚酸酯1.0g/d。结果:患者移植后均获得造血重建,中性粒细胞>0.5×109/L和血小板>20×109/L的中位时间分别是12.5(10～14d)和22d(18～25d)。4例患者发生急性Ⅰ度GVHD,其中1例2个位点不相合者进展为急性肠道和肝脏Ⅳ度GVHD,于+81d合并感染死亡。1例发生迟发性出血性膀胱炎。中位随访时间20个月(5～25个月)。无病存活3例,其中2例存活在1年以上。结论:供者应用GCSF后采髓,多种免疫抑制剂联合应用的HLA不全相合未去除T细胞骨髓移植,在治疗慢性粒细胞白血病过程中,有效地降低了急性重症GVHD发生,提高了无病生存。     

3例复发难治性B细胞急性淋巴细胞白血病的治疗及移植物抗宿主病预防

目的 通过对3例复发难治性B细胞急性淋巴细胞白血病(B-ALL)患者的治疗方法及移植治疗后移植物抗宿主病(GVHD)的预防方法分析,总结这类疾病有效治疗方案及预防方案.方法 3例复发难治性B-ALL患者中2例为费城染色体Ph(+),在确定半相合供者后,给予CD19靶向嵌合抗原受体T(CAR-T)细胞桥接治疗,缓解后接受亲缘半相合异基因造血干细胞移植(haplo-HSCT),采用造血干细胞移植方案GIAC进行移植治疗,同时在移植后给予小剂量环磷酰胺(PTCY,14.5 mg/kg体质量)预防GVHD.结果 3例B-ALL患者CAR-T细胞治疗后均获得了完全缓解(CR),其中2例达分子生物学缓解,1例达流式MRD阴性缓解;其中2例患者顺利接受haplo-HSCT,回输后均成功供者细胞造血重建,无严重GVHD,白血病疾病状态稳定.另1例伴有p.T315I突变的Ph(+)-ALL患者,CAR-T细胞治疗后获得短暂分子生物学缓解,但在移植准备过程中再次复发后失去移植机会.结论 CAR-T治疗后序贯亲缘半相合移植治疗复发难治性B-ALL患者效果良好;小剂量PTCY联合GIAC移植方案可以安全有效地预防GVHD的发生;CAR-T细胞治疗后短期再次复发的B-ALL患者目前仍然治疗困难.     

异基因造血干细胞移植术后供体型复发及文献复习

目的：提高对异基因造血干细胞移植术后供体型复发的认识水平，分析其发生原因。方法：回顾性分析2001年9月～2003年12月在我院行异基因造血f：细胞移植术后供者复发3例的移植前状态、治疗和预后。结果：3例患者移植后全部重建造血，DNA序列分析表明1个月内移植物植入．复发时嵌合体检查为完全供体型。结论：移植前未达完全缓解．或反复复发的白血病患者移植后易复发；STR-PCR结果显示．患者可在完全供者嵌合体状态下复发。     

异基因造血干细胞移植治疗白血病76例临床观察

目的观察不同来源的异基因造血干细胞移植治疗白血病的疗效并探讨主要并发症的处理方案。方法对2001年9月至2007年3月第四军医大学西京医院血液科76例白血病患者行异基因造血干细胞移植治疗,其中慢性粒细胞白血病34例,急性髓性白血病24例,急性淋巴细胞白血病15例,T细胞淋巴瘤/白血病3例。人类白细胞抗原(HLA)全相合的同胞供者57例,1个HLA位点不合同胞供者3例,HLA单倍型半相合同胞供者7例,非血缘供者9例。预处理方案采用改良的马利兰联合环磷酰胺(BUCY)或改良的环磷酰胺联合全身放疗及阿糖胞苷或鬼臼乙叉甙(CyTBI Ara-c/VP-16)方案。采用标准的环孢素A(CsA)联合短期甲氨蝶呤(MTX)方案预防移植物抗宿主病(GVHD);无关供者移植加用抗人胸腺细胞球蛋白,单倍型半相合移植同时加用CD25单克隆抗体。结果96.1%(73/76)获得植入。24.7%(18/73)出现急性GVHD,32.9%(24/73)出现慢性GVHD;合并重症肝静脉闭塞病2例;并发纯红细胞性再生障碍性贫血5例。随访3~72个月,现存活56.6%(43/76),43.4%(33/76)在移植后1~36个月时死亡,19例死于白血病复发,14例死于移植相关并发症。结论多种来源的异基因造血干细胞移植是治疗白血病的有效方法,于慢性粒细胞白血病慢性期、急性白血病缓解期移植效果较好,移植前处于高危难治状态的病例复发率仍较高。     

亲属间HLA半相合干细胞移植治疗难治复发性白血病患者30例疗效分析

目的观察亲属间HLA半相合干细胞移植(SCT)治疗难治复发性白血病(RL)的疗效。方法对自1998年8月至2004年8月期间进行HLA半相合SCT治疗30例RL的治疗结果进行分析。结果30例患者均为RL,其中急性非淋巴细胞白血病13例,急性淋巴细胞白血病10例,慢性粒细胞白血病6例,Ⅳ期淋巴瘤1例。男18例,女12例,中位年龄25(3~52)岁。30例患者均接受HLA半相合SCT。12例为父母给子女,4例为子女给父母,其余为同胞。HLA3个位点不合12例,HLA2个位点不合13例,5例HLA1个位点不合。预处理方案为氟达拉宾、马利兰、环磷酰胺方案,部分患者在此方案基础上加用抗人胸腺细胞球蛋白。平均移植单个核细胞数5·0(2·9~8·0)×108/kg,CD3+4细胞数为5·5(3·0~6·5)×106/kg。30例中24例完全供者植入,3例供、受者部分嵌合,经供者淋巴细胞输注转为完全供者型;1例移植失败,1例移植后2d死于重症霉菌感染,1例移植后28d死于重症肝静脉阻塞病;移植后WBC>1·0×109/L平均时间14(11~18)d,血小板>20×109/L时间15(11~18)d。6例发生Ⅲ~Ⅳ度重症移植物抗宿主病(GVHD),7例发生慢性GVHD,白血病缓解率90%,随访3~60个月,平均生存时间为16·9个月。7例复发,其中2例脑膜白血病复发,复发中位时间为10(3~24)个月,14例生存,10例仍无病生存。结论HLA半相合SCT能使大部分RL患者缓解,也能使部分RL患者获长期生存;HLA半相合SCT具有较强的移植物抗白血病效应,但在疾病状态下进行HLA半相合SCT,白血病仍会复发。     

半相合淋巴细胞输注治疗急性白血病研究进展

供体淋巴细胞具有移植物抗白血病 (ＧＶＬ)效应 ,而ＧＶＬ效应主要由异体Ｔ细胞介导。目前 ,半相合供者淋巴细胞输注 (ＤＬＩ)已成为治疗异基因造血干细胞移植 (ａｌｌｏ ＨＳＣＴ)后慢性髓细胞白血病(ＣＭＬ)复发的最有效方法 ,并逐渐用于其他恶性血液病ａｌｌｏ ＨＳＣＴ后复发的治疗 ,取得了令人瞩目的效果 ,应用范围逐渐扩展到ａｌｌｏ ＨＳＣＴ的非复发并发症和未经骨髓移植预处理患者的过继免疫治疗。而半相合ＤＬＩ治疗急性白血病成为本领域研究的热点。本文对半相合ＤＬＩ治疗的效应机制 ,临床应用的并发症以及时机、剂量等方面的研…     

供者用粒细胞集落刺激因子单倍体骨髓移植的临床研究

目的探讨供者用粒细胞集落刺激因子(G-CSF)和受者联合应用多种免疫抑制剂治疗的单倍体骨髓移植在降低重症移植物抗宿主病(GVHD)和改善无病生存的疗效.方法单倍体骨髓移植治疗白血病13例(单倍体移植组),移植后结果和连续完成的13例白血病HLA匹配异基因移植 (相合移植组) 相比较,单倍体移植方法是供者应用G-CSF 250 μg/d,连用7 d后采髓, 受者GVHD预防除环孢素A(CSA)和甲氨蝶呤(MTX)外,在移植前4～1 d用抗胸腺细胞球蛋白(ATG) 5 mg*kg-1*d-1, 移植后7 d始加服霉酚酸酯(MMF).结果单倍体移植组植入物CD+34细胞中位数6.1×106/kg,是相合移植组输入CD+34细胞中位数2.5×106/kg的2倍多(P＜0.01),单倍体移植组和相合移植组植入物CD+3细胞中位数分别是50.5×106/kg和47.0×106/kg(P＞0.05).移植后无1例发生植入失败,两组造血重建速度无差异(P＞0.05),所有患者经骨髓植活直接证据检测证实为完全供者造血.单倍体移植组发生急性Ⅱ～Ⅳ GVHD 5例(38.5%),可评价的8例中7例发生慢性GVHD(87.5%),为局限性慢性GVHD,这与相合移植组差异无显著性 (P＞0.05).单倍体移植组中位随访453 d(180～690),移植相关死亡5例,无复发死亡病例,剩余8例无病存活(61.5%).相合移植组中位随访510 d(220～810),移植相关死亡2例,复发死亡2例,9例无病存活(69.2%),通过比较两组差异无显著性(P＞0.05).结论本研究单倍体骨髓移植治疗白血病是一种安全和有效方法,在降低重症急性GVHD 及改善无病生存方面和HLA相合同胞供者移植相当.     

HLA半相合非清髓异基因造血干细胞移植治疗难治性急性白血病1例

目的探讨HLA半相合非清髓异基因造血干细胞移植(NAST)在治疗难治性急性白血病中的作用。方法2002-06采用非清髓预处理的NAST治疗军事医学科学院附属307医院难治性急性髓性白血病患者1例。预处理方案主要由抗淋巴细胞球蛋白(ATG)、阿糖胞苷(Ara-C)、氟达拉滨(Flu)和环磷酰胺(CTX)等组成。移植物抗宿主病(GVHD)的预防采用环孢素A(CSA)、霉酚双酯(MMF)、甲氨蝶呤(MTX)和CD25单抗。结果患者移植过程顺利,于移植后第100d转为完全供者型植入。患者于移植后出现皮肤Ⅰ度GVHD,经治疗后好转。结论应用HLA半相合NAST治疗难治性急性白血病患者,简便安全,疗效好,为无HLA相合供者的白血病患者治疗开辟了新的治疗手段。     

Restriction fragment length polymorphism analysis of hematopoietic cells following successful treatment of relapsed acute lymphoblastic leukemia following bone marrow transplantation

Despite aggressive therapy for leukemia in the form of bone marrow transplantation (BMT) relapse occurs in a significant number of cases. The origin of the leukemic relapse, whether it is of donor or recipient origin, and how best to treat the patients continue to pose problems for the clinician. In this paper we present a case in which the cytogenetics suggested that the relapse was of donor origin; however, molecular analysis revealed that the leukemic population was of host origin. The leukemic relapse following the BMT was treated with a second BMT. This resulted in a remission of 28 months after which leukemic relapse was again diagnosed. Using conventional chemotherapy it was possible to obtain another complete remission. This case illustrates a pitfall to cytogenetic analysis and two contrasting methods of dealing with leukemic relapse following BMT.     

Allogeneic bone marrow transplantation for high-risk acute lymphoblastic leukemia during first complete remission

Fifty-three patients with high-risk acute lymphoblastic leukemia (ALL) under age 50 with a histocompatible sibling donor received high-dose radiochemotherapy followed by allogeneic bone marrow transplantation (BMT). The high-risk factors used to identify the patients were: white blood cell count at initial presentation, cytogenetic abnormalities, age, extramedullary leukemic infiltration, and time from initial therapy to complete remission. Patients with one or more of the above risk factors who received BMT have a disease-free survival of 61% with a median follow-up of 66 months (range 11 months to 10.6 years), and an actuarial relapse rate of 10%. This study demonstrates that patients with high-risk ALL achieve a significant disease-free survival and cure rate with the use of allogeneic fully matched sibling BMT. However, a properly designed prospective study comparing the outcome of BMT with the best currently available chemotherapy data is required to define the ultimate role of BMT in this group of patients.     

Molecular analysis of relapse in chronic myeloid leukemia after allogeneic bone marrow transplantation

Four patients with Philadelphia (Ph') positive chronic myeloid leukemia (CML) were studied before, after, and on relapse following allogeneic bone marrow transplantation (BMT). Southern analysis of DNA from cells collected before and at relapse after BMT was performed in order to investigate the origin of the leukemia at relapse. Using minisatellite probes we showed that the relapse occurred in cells of host origin in all four patients and this was confirmed with a Y chromosome specific probe in two male patients who had a female donor. Furthermore, using two probes for the breakpoint cluster region (bcr) on chromosome 22, we showed that leukemic cells at relapse bore identical rearrangements to those in the disease at time of presentation of each patient. We conclude that relapse in all four patients is due to re-emergence of the original leukemic clone.     

The analysis of leukemic relapse after allogeneic bone marrow transplantation]

This report presents the analysis of leukemic relapse of 52 patients who received allogeneic bone marrow transplantation between July 1984 and May 1990. Conditioning regimen consisted of TBI + CY and GVHD prophylaxis consisted of cyclosporin-A and methotrexate. The relapse ratios of chronic myelogenous leukemia (CML) (21 in chronic phase, 1 in accelerated phase, 1 in blastic crisis), acute nonlymphocytic leukemia (ANLL) (all 17 in 1st CR), acute lymphocytic leukemia (ALL) (all 12 in 1st CR) were 13%, 18%, 25%, respectively, and 3 year disease free survival (DFS) was as follows, CML 68%, ANLL 72%, ALL 49%. Regarding acute GVHD grading and chronic GVHD presence, 3 year DFS was as follows, acute GVHD 0 degree: 59%, I degree: 78%, II degree-IV degree: 53%, chronic GVHD (+): 82% GVHD (-): 77%. In our center leukemic relapse has been the major cause of death after BMT since 1984. Among 9 relapsed cases, one recurred more than 3 years after BMT, and another one got recurrent leukemia of donor origin.     

Humanized anti-CD25 monoclonal antibody for prophylaxis of graft-vs-host disease (GVHD) in haploidentical bone marrow transplantation without ex vivo T-cell depletion

OBJECTIVE: To investigate the effects of a novel anti-IL-2 receptor (CD25) monoclonal antibody, basiliximab, on graft-vs-host disease (GVHD) and engraftment in haploidentical bone marrow transplantation (BMT). MATERIALS AND METHODS: Thirteen consecutive high-risk leukemia patients (age 9-41) underwent haploidentical BMT with G-CSF-primed marrow as stem cells without ex vivo T-cell depletion. Basiliximab, along with a combination of cyclosporine (CSA), methotrexate (MTX), and mycophenolate mofetil (MMF), was used for GVHD prophylaxis. Immunophenotyping, limited-dilution assay, and colony-forming assays were used to measure the effect of basiliximab on the subsets of lymphocytes, cytotoxic T-lymphocyte precursors (CTLp), and hematopoietic cells. RESULTS: All patients established successful trilineage engraftment with full donor chimerism. No patients developed grade II-IV acute GVHD. Patients who survived more than 12 months and were free of relapse showed limited chronic skin GVHD. Ten of 13 patients are currently alive with a Karnofsky performance score of 100% at median follow-up of 17 months (range 12-24 months). Basiliximab significantly decreased alloreactive CTLp by 10-fold to 100-fold in limiting-dilution assays. It had no effect on hematopoietic stem and progenitor cells as determined by in vitro colony-forming assays. CONCLUSION: The addition of basiliximab to CSA, MMF, and MTX as GVHD prophylaxis effectively reduced severe lethal GVHD in haploidentical BMT. It is possible to selectively eliminate or reduce the number of alloreactive T cells with anti-CD25 antibody, which results in prevention of or a reduction in the severity of GVHD.     

Allogeneic bone marrow transplantation versus chemotherapy in the treatment of childhood acute lymphoblastic leukemia in second complete remission

Seventy-six patients between the ages of 2 and 17 years with acute lymphoblastic leukemia (ALL) achieved a second complete remission induced by polychemotherapy. Twenty-one had an HLA-identical donor and underwent allogeneic bone marrow transplantation (BMT) after conditioning with total body irradiation and cyclophosphamide. The remaining 55 patients lacked a suitable donor and received intensive chemotherapy as treatment. Fifteen patients were excluded from the analysis because they relapsed within 3 months after achieving a second complete remission. Three of the 21 BMT patients died of transplant-related complications and seven relapsed between 90 and 480 days after transplantation. Eleven patients are alive and disease free at 5.5-71 months with an actuarial survival of 47.1%; eight patients are on a plateau extending from 22 to 71 months. Thirty-three patients treated with chemotherapy died from relapse and seven are alive and disease free 7.5-99 months from the second remission, with an actuarial survival of 9%. The probability of survival was significantly higher in the BMT group (p less than 0.025). The probability of remaining in complete remission in the BMT group was 58.5% versus 10.9% in the chemotherapy group (p less than 0.005). Our results show that BMT is the best alternative therapy for children affected by ALL who have had a relapse in the marrow.     

Characterization of mixed chimerism in patients with chronic myeloid leukemia transplanted with T-cell-depleted bone marrow: involvement of different hematologic lineages before and after relapse

We have characterized mixed chimerism (MC) in five patients with chronic myeloid leukemia (CML) who received transplants with T-cell- depleted bone marrow (BM) and who relapsed within 4 years after transplantation. To study the possible relation of MC with relapse, we purified different populations of leukocytes and analyzed their donor/recipient origin by a method based on polymerase chain reaction amplification of minisatellite DNA regions. Our results show that before relapse, all hematopoietic recipient cells are T cells, whereas monocytes, B, and natural killer (NK) cells are of donor origin. This observation does not appear to be specific for CML as similar results were found in two control patients with acute myeloid leukemia (AML). At the time of (CML) relapse, recipient granulocytes, monocytes, and erythrocytes appeared and progressively replaced the respective lineages of donor origin. No other lineages seemed to be involved as B cells and NK cells remained of donor origin and no significant changes in the number of recipient T cells were detected. In this respect relapse of CML after BM transplantation (BMT) seems not to be very different from the primary disease in chronic phase before transplantation. Furthermore, we conclude that after BMT, an association between mixed chimerism before relapse and the (CML) relapse does exist because both phenomena are consequences of T-cell depletion of the BM graft. However, this correlation might well be indirect as the MC caused by the recipient T cells appears to be independent of the one caused by the recurrent disease.     

Successful graft-versus-leukemia effect of second bone marrow transplantation on relapsed leukemia cutis that was refractory to intensive chemotherapy and donor lymphocyte transfusions in a patient with acute monocytic leukemia

We report a patient with acute monocytic leukemia (AMoL; M5) who received a second bone marrow transplantation (BMT) with graft-versus-leukemia (GVL) effect on relapsed leukemia cutis, which had been refractory to intensive chemotherapy and donor lymphocyte transfusions (DLTs). A 21-year-old woman was diagnosed with AMoL and achieved complete remission after intensive chemotherapy. The patient received a nonmanipulated allogeneic BMT from her HLA-identical father. Skin tumors developed in her upper extremities, chest, and thigh 11 months after BMT. Leukemia cutis was confirmed by skin biopsy. There was no evidence of relapse in bone marrow. The patient received several courses of chemotherapy and DLTs for the skin relapse, but the skin tumors persisted. The patient then received a second BMT from the same donor. On day 80, grade II acute graft-versus-host disease developed, and the remaining skin tumors were eradicated on day 98, most probably because of GVL effect.     

Treatment of infant leukemia with busulfan, cyclophosphamide +/- etoposide and bone marrow transplantation.

Infants with acute leukemia have a poor chance of being cured by conventional chemotherapy. We therefore treated cases of infant leukemia with high dose chemotherapy followed by bone marrow transplantation (BMT). Six suffered from acute leukemia and one from refractory anemia with excess of blasts (RAEB-t). The conditioning regimen consisted of busulfan (BU) and cyclophosphamide (CY), and was intensified by adding etoposide (VP) in four cases. At the time of BMT the children were 4, 5, 12, 13, 13, 14, and 20 months old. Three children were autografted, three received HLA-identical marrow from a sibling donor, and one child received matched unrelated donor marrow. All five children who were grafted in complete (CR) or partial remission (PR) are alive and well in CR 7, 13, 24, 37, and 46 months after allogeneic (two patients) or autologous (three patients) BMT, and 13, 17, 29, 42, and 53 months after initial diagnosis. The child with RAEB-t and the one transplanted in second chemotherapy-resistant relapse of acute non-lymphoblastic leukemia relapsed at 7 and 17 months respectively. The chemotherapy regimen was well tolerated. BU-CY-VP is a promising alternative treatment to regimens including total body irradiation for very young children suffering from acute leukemia.