Attenuation of GVHD for Allo-Bone Marrow Transplantation Recipient by FasL-Fas Pathway in an H-2 Haplotype Disparate Mouse Combination

In order to explore a new special and effective way to prevent graft versus host disease (GVHD) after allogenic bone marrow transplantation (allo-BMT), the stem cell antigen-1 (Sca-1) + early hematopoietic cells (EHC) from BALB/c mouse (H-2 d) were introduced with exogenous mouse Fas ligand (mFasL) cDNA gene by the retrovirus-mediated gene transfer and expanded for one week, and then they were co-cultured with the spleen mononuclear cells (SMNC) from BAC mouse (H-2 d×b) as one way mixed lymphocyte reaction (OWMLR). The cytotoxicity of treated BAC mouse SMNC against Na 2 51CrO 4 labeling SMNC from BALB/c mouse was observed. The bone marrow mononuclear cells (BMMNC) from BAC mouse treated by the above methods were transplanted into lethally-irradiated congenic BALB/c mice to observe the occurrence of GVHD. The results showed that the SMNC from BAC mouse after OWMLR with exogenous mFasL cDNA gene-transduced hematopoietic cells (HC) from BALB/c mouse in a ratio of 1 to 5 exhibited an obvious inhibition of the cytotoxicity against the BALB/c mouse spleen cells at different effector/target ratios as compared to the control group (P<0.01). The gradeⅠ GVHD or no GVHD and the 80 % survival rate at day 60 post-BMT were observed in the BALB/c mouse receiving BAC mouse BMMNC treated with similar way, while the grade Ⅱ-Ⅲ GVHD and the 20 % survival rate were noted in the control group (P<0.01). It is suggested that the attenuation of GVHD in allo-BMT recipient could be successfully achieved through FasL-Fas pathway in an H-2 haplotype disparate mouse combination.

After allogenic bone marrow transplantation agent of hemorrhagic cystitis: BK virus

Hemorrhagic cystitis is a common and in its severe form potentially life threatening complication of hematopoietic stem cell transplantation. Hemorrhagic cystitis is defined as a diffuse inflammatory condition of the urinary bladder due to an infectious or noninfectious etiology resulting in bleeding from the bladder mucosa. Hemorrhagic cystitis is characterized by lower urinary tract symptoms including dysuria, hematuria and hemorrhage. The most common cause is a bacterial infection that usually responds promptly to treatment. But chronic and recurrent hemorrhagic cystitis often arises from anticancer chemotherapy or radiotherapy for the treatment of pelvic malignancies. Infectious etiologies are less common causes of chronic hemorrhagic cystitis except in immunocompromised hosts like bone marrow transplant recipients. Hemorrhagic cystitis is a significant complication of bone marrow transplantation which influences economic and survival outcome. Hemorrhagic cystitis can be divided into two classes according to onset time;early and late onset time. Earlyonset hemorrhagic cystitis is commonly associated used with chemo-radiotherapy protocols in some of the preparatory regimens. More than one factor is accused in the etiology of late onset hemorrhagic cystitis. Here, we present a patient whose hematuria started after 54 days from allogeneic stem cell transplantation.

Cost Effectiveness Analysis of Filgrastim versus Placebo in Post AIIogentic Bone Marrow Transplantation

Filgrastim is used to accelerate hematopoietic recovery after ABMT （allogeneic bone marrow transplantation）. Its impact on the total cost of patient care remains to be explored. We therefore undertook a cost effectiveness analysis in the context of a randomized single blinded clinical trial of Filgrastim versus placebo in post ABMT. A primary endpoint, duration of myelosuppression, and three secondary end points （number of days of fever, length of hospital stay, survival at one hundred days） were used to assess efficacy. Direct costs were evaluated and allowed the calculation of the ICER （incremental cost-effectiveness ratios） for the major endpoint of the trial. Sixteen patients were included in the study. The duration of myelosuppression was significantly decreased in the Filgrastim arm with medians of 15 days vs. 19 days in the placebo arm （p = 0.023）. Cost analysis showed no statistically significant difference between the two arms. According to the calculation of ICER, Filgrastim was more costly and more effective than placebo for the number of days of aplasia avoided and the number of days with fever avoided. Placebo strictly dominated filgrastim for days of hospitalization avoided. Filgrastim has proven effective in reducing the duration of aplasia without increasing costs.

An improved anti-leukemic effect achieved with donor progenitor cell infusion for relapse patients after allogeneic bone marrow transplantation

Objective To observe the antileukemic effect in relapse patients by infusion of donor immunocompetent cells with or without granulocyte colony-stimulating factor (G-CSF) mobilization.Methods Twenty patients with leukemia in relapse after allogeneic bone marrow transplantation (allo-BMT) were treated with chemotherapy followed by donor-derived lymphocytes (DDL) without G-CSF mobilization (Group A, n=11), or donor peripheral blood progenitor cells (PBPCs) with G-CSF mobilization (Group B, n=9).Results Five patients in Group A were in hematologic relapse. After DDL infusion, 3 of 5 patients had a temporary complete remission (CR) and relapsed after 3, 7 and 10 months, respectively. One achieved partial remission and died of interstitial pneumonia; and the other one showed no response. Another 6 patients in Group A were in cytogenetic relapse or central nerve system (CNS) leukemia, and all achieved CR and remained in disease free survival (DFS) for 10 to 98 months after DDL infusion. All 9 patients in group B were in hematologic relapse. Three patients with chronic myeloid leukemia (CML) had cytogenetic and molecular remission for 16, 35 and 51 months, respectively after PBPC infusion; and 5 patients with acute lymphoid leukemia (ALL) had CR and were still in CR for 10 to 18 months except 1 patient relapsed soon. And the other one with AML showed no response to the therapy.Conclusion Donor immunocompetent cells infusion is an effective therapy for relapsed leukemia after allo-BMT, especially for the patients with early (molecular and cytogenetic) or CNS relapse. Infusion of donor PBPC mobilized by G-CSF seems to have more potentiated graft-versus-leukemia (GVL) effect than DDL infusion.

Graft-versus-leukemia effects from donor lymphocyte infusion after nonmyeloablative allogeneic bone marrow transplantation in mice

BABSTRACTackground Nonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).Methods A total of 615 (H-2~k) mice were injected with L615 tumor cells and received 500 cGy (^(60)Coγ-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3×10~7 bone marrow cells and 1×10~7 spleen cells from BALB/C (H-2~d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells (2×10~7) on day 14 or 21, or donor spleen cells (5×10~7) pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.Results The median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P<0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.Conclusion Donor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.

Influence of the degree of donor bone marrow hyperplasia on patient clinical outcomes after allogeneic hematopoietic stem cell transplantation

This study evaluated the influence of the degree of donor bone marrow(BM)hyperplasia on patient clinical outcomes after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Twelve patients received allo-HSCT from hypoplastic BM donors between January 2010 and December 2017.Forty-eight patients whose donors demonstrated BM hyperplasia were selected using a propensity score matching method(1:4).Primary graft failure including poor graft function and graft rejection did not occur in two groups.In BM hypoplasia and hyperplasia groups,the cumulative incidence(CI)of neutrophil engraftment at day 28(91.7%vs.93.8%,P=0.75),platelet engraftment at day 150(83.3%vs.93.8%,P=0.48),the median time to myeloid engraftment(14 days vs.14 days,P=0.85)and platelet engraftment(14 days vs.14 days,P=0.85)were comparable.The 3-year progression-free survival,overall survival,CI of non-relapse mortality and relapse were 67.8%vs.71.7%(P=0.98),69.8%vs.77.8%(P=0.69),18.5%vs.13.6%(P=0.66),and 10.2%vs.10.4%(P=0.82),respectively.In multivariate analysis,donor BM hypoplasia did not affect patient clinical outcomes after allo-HSCT.If patients have no other suitable donor,a donor with BM hypoplasia can be used for patients receiving allo-HSCT if the donor Complete Blood Count and other examinations are normal.

Relapse of acute lymphoblastic leukemia in the pancreas after allogeneic bone marrow transplantation

The occurrence of metastatic lesions in the pancreas of patients with cancer, including hematological cancers,is uncommon （1.6%-37.5%） and of these, the majority of patients will have widespread disease.1-7 Isolated potentially resectable pancreatic metastases are rarely seen. Resection of metastatic tumors of the pancreas has been reported, but its role in improving survival rates or quality of life is not clear.7,8 However,

FILGRASTIM (G-CSF) FOR ALLOGENEIC BONE MARROW TRANSPLANTATION

Conditioning for allogeneic bone marrow transplantation causes severe neutropenia until new

TREATMENT OF APLASTIC ANEMIA: ALLOGENEIC BONE MARROW TRANSPLANTATION VERSUS IMMUNOMODULATION THERAPY WITH ANTILYMPHOCYTE GLOBULIN

There are two arms in the management of aplastic anemia,one allogeneic bone marrow transplantation(BMT) and the other immunomodulation therapy with antilymphocyte globulin(ALG).

THE ROLE OF ETOPOSIDE (VP-16) IN PREPARATORY REGIMENS FOR PATIENTS WITH LEUKEMIA UNDERGOING ALLOGENEIC BONE MARROW TRANSPLANTATION

In 1983,we began a series of five clinical trials with the goal of reducing the relapse rate following

A comparative study of unrelated donor and matched-sibling donor allogeneic hematopoietic stem cell transplantation in children and adolescents with acquired severe aplastic anemia

Objective To evaluate the efficacy of unrelated do-nor allogeneic hematopoietic stem cell transplantation(URD allo-HSCT).for children and adolescents with severe aplastic anemia(SAA).Methods Clinical data of34 SAA children and adolescents undergoing allo-HSCT were retrospectively analyzed from October 2001 to October 2015.According to the source of donor,the

小剂量骨髓联合外周血造血干细胞移植治疗重型再生障碍性贫血18例临床研究

目的:探讨小剂量骨髓联合外周血造血干细胞(PBSC)移植治疗重型再生障碍性贫血(SAA)的疗效及安全性。方法:纳入以小剂量骨髓(中位体积200 mL)联合PBSC移植治疗的18例SAA患者的临床资料,回顾分析其造血重建情况、移植物抗宿主病(GVHD)发生率、感染发生率及生存率等指标。结果:18例患者的中位年龄27(13~52)岁,男11例,女7例;SAA-Ⅰ型15例,SAA-Ⅱ型3例;单倍体相合供者15例,同胞全相合供者3例;预处理:氟达拉滨+环磷酰胺+抗人胸腺球蛋白方案2例,氟达拉滨+环磷酰胺+移植后环磷酰胺方案8例,氟达拉滨+环磷酰胺+抗人胸腺球蛋白+移植后环磷酰胺方案8例;采用环孢素或他克莫司+短疗程甲氨蝶呤联合吗替麦考酚酯预防GVHD。所有患者均获得造血重建,仅1例患者出现继发性移植物功能不良;10例(55.6%)出现总计19例次移植后感染,其中10例次为巨细胞病毒(CMV)或EB病毒(EBV)血症,6例次细菌感染;急性GVHD发生率为16.7%,其中Ⅰ度1例,Ⅱ度2例;慢性GVHD发生率为16.7%,均为轻度;中位随访26(2~64)个月,移植后2年总生存率为72.2%(95%CI:51.4~93.0)。5例患者死亡,4例死于肺部感染,1例死于颅内感染。结论:采用小剂量骨髓联合PBSC是SAA患者进行造血干细胞移植的有效且可行的移植物选择策略。

Outcomes of peripheral blood stem cell transplantation in patients from human leukocyte antigen matched or mismatched unrelated donors

Background AIIogeneic peripheral blood stem cell transplantation from unrelated donors （UR-PBSCT） is an alternative treatment for many hematologic diseases due to lack of human leukocyte antigen （HLA）-identical sibling donors. This study aimed to evaluate the impact of the degree of the HLA match on the clinical efficacy of UR-PBSCT. Methods Patients who underwent UR-PBSCT from September 2003 to September 2012 were retrospectively investigated. They were divided into three groups according to high-resolution molecular typing. SPSS version 17.0 was used to analysis and compare the statistics of engraftment, incidence of GVHD, other complications and survival among the groups. Results One hundred and eleven patients received UR-PBSCT, 60 of them with an HLA matched donor （10/10）, 36 of them with a one locus mismatched donor （9/10）, and 15 of them with a two loci mismatched donor （8/10）. Similar basic characteristics were found in the three groups. No differences were found in engraftment of myeloid cells or platelets in the three groups （P〉0.05）. Two-year cumulative incidence of relapse, overall survival （OS） and disease-free survival （DFS） among those three groups were similar （P〉0.05）. The cumulative incidence of 100-day Ill-IV aGVHD in the HLA matched group and the one HLA locus mismatched group were significantly lower than that in the two HLA loci mismatched group （3.3%, 8.6%, and 26.7%, P=0.009）. The occurrence rate of new pulmonary infections in the HLA matched group was lower than in the two HLA mismatched groups （26.67%, 52.78%, and 41.18%, P=0.035）. The cumulative incidence of 100-day and 2-year transplantation related mortality （TRM） in two HLA loci mismatched group was higher than in the HLA matched group and in the one HLA locus mismatched group, （8.4%, 11.8% and 33.3%, P=0.016） and （12.3%, 18.7% and 47.5%, P=0.002）. Conclusions HLA mismatch will not significantly impact the engraftment or 2-year survival after UR-PBSCT, but two mismatched HLA loci may increase the cumulative incidence of severe aGVHD and TRM. Chin Med J 2014;127 （14）： 2612-2617

大鼠骨髓间充质干细胞对同种异体骨髓移植造血重建的影响

为了探讨大鼠骨髓间充质干细胞(mesenchymalstemcell,MSC)对同种异体骨髓移植造血重建的影响,建立大鼠同种异体骨髓移植模型,通过外周血像检测、病理分析和流式细胞术检测综合评价MSC对骨髓移植(bonemarrowtransplantation,BMT)的积极作用。结果表明:①共移植后30天,MSC可在致死量照射的受者存活,并被发现受者胸腺、脾脏和骨髓;②MSC可促进BMT后造血重建,促进外周血白细胞、淋巴细胞和血小板的明显恢复;促进骨髓细胞数恢复及B淋巴系、巨核系分化增强;有利骨髓组织学的明显恢复发善。结论:大鼠MSC与骨髓共移植可在受体的造血器官长期存在,MSC可明显促进同种异体骨髓移植后造血重建。

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/Th2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/Th2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。

G-CSF动员的异基因外周血干细胞和骨髓联合移植治疗白血病

为了探讨粒细胞集落刺激因子(G-CSF)动员的异基因外周血干细胞和骨髓联合移植治疗白血病的疗效, 给予供者G-CSF 8-10μg／(kg·d),皮下连续注射5天,以动员外周血和骨髓干细胞,在第4,5天采集外周血干细 胞,第7天采集骨髓;预处理采用氟达拉宾,白消安和环磷酰胺组成的新方案。结果表明,12例全部成功植入,中性 粒细胞>0.5×109／L的时间为10.2(9-12)天,血小板>20×109／L的时间为12.5(12-14)天。移植后观察100 天,4例(33.3％)发生Ⅱ度急性移植物抗宿主病(aGVHD),10例出现慢性移植物抗宿主病(cGVHD),中位随访时 间5个月,2例死亡,余10健康生存。结论:G-CSF动员的和外周血造血干细胞骨髓联合移植是安全有效的治疗方 法,可以获得长期稳定的植活并不增加aGVHD和cGVHD等并发症。

马利兰和环磷酰胺预处理方案的非亲缘异基因骨髓移植治疗骨髓增生异常综合征

目的:评估以马利兰和环磷酰胺(Bu-CY2)为预处理方案的非亲缘异基因骨髓移植治疗骨髓增生异常综合征(MDS)的临床疗效。方法:对6例MDS患者进行非亲缘异基因骨髓移植术,以Bu-CY2为预处理方案,Bu 4 m g.k-g 1.d-1,-7 d^-4 d,CY 60 m g.k-g 1.-d 1,-3 d^-2 d。输入单个核细胞数(MNC)为3.38×108/kg(2.4×108/kg^4.6×108/kg),CD 34+细胞数5.81×106/kg(1.2×106/kg^8.5×106/kg),粒-巨噬细胞集落形成单位(CFU-GM)数2.88×105/kg(1.61×105/kg^4.56×105/kg)。以霉酚酸酯加环孢素A(C sA)和短程氨甲蝶呤(MTX)预防移植物抗宿主病,前列腺素E1脂质微球预防肝静脉阻塞病。结果:6例患者中性粒细胞≥0.5×109/L的中位时间为18 d(13~21 d),血小板≥20×109/L的中位时间为21 d(13~24 d)。经DNA短串联重复序列多态性分析和染色体检查,均为供者骨髓植活。早期死亡率为0,移植后随访时间为27个月(6~60个月)。目前实际无病生存6例,缓解期实际生存率100%。结论:以Bu-CY2为预处理方案的非亲缘异基因骨髓移植是治疗MDS的有效方法。

MHC半相合骨髓移植后慢性GVHD小鼠模型的建立

目的建立MHC半相合异基因骨髓移植(allo-BMT)后慢性GVHD小鼠模型。方法以(Balb/c×C57BL/6)F1H-2d/b(CB6F1)雌性小鼠为受者,预处理条件为不同剂量的全身照射(TBI,60Co照射),输注MHC半相合Balb/cH-2d雄性小鼠骨髓细胞与不同数量脾细胞,观察移植后小鼠体质量变化,靶器官病理变化,血清抗ssDNA抗体、抗dsDNA抗体。结果在照射剂量为8Gy,输注骨髓细胞数量为8×106、脾细胞数量为4.5×107的小鼠至实验结束(移植后100d)全部存活,体质量减轻与对照组和其它实验组相比差异显著(P<0.05)。血清抗ssDNA抗体、抗dsDNA抗体较对照组和其它实验组显著升高(P<0.05)。该组皮肤、肝脏等病理改变明显。结论在照射剂量为8Gy,输注骨髓细胞数量为8×106、脾细胞数量为4.5×107的小鼠成功诱导出半相合allo-BMT慢性GVHD,为进一步研究慢性GVHD的发病机理、生物学特性、干预因素等打下了重要基础。

同种异体骨复合自体骨髓干细胞移植治疗良性骨肿瘤和瘤样病变

背景:同种异体骨是临床常用的骨移植材料,但缺乏诱导成骨能力是最大的问题。目的:评价良性骨肿瘤及瘤样病变刮除或切除后应用同种异体骨复合自体骨髓干细胞修复骨缺损的效果。方法:65例良性骨肿瘤(包括瘤样病变)患者,根据植骨情况分为2组。复合骨髓干细胞植骨组35例患者根据预计植骨量从每位患者两侧的髂前上棘或髂后上棘抽取红骨髓20-40mL,经体外分离、纯化、培养扩增骨髓基质干细胞备用,在植骨前将同种异体骨颗粒与骨髓基质干细胞充分混匀。肿瘤刮除或切除后,将混匀的骨髓基质干细胞与同种异体骨颗粒,植入骨缺损区内。单纯植骨组将用生理盐水浸泡半小时的同种异体骨植入骨缺损区内。分别于治疗后1,3,6,12个月进行植骨区X射线检查,比较两组病例同种异体骨颗粒界限模糊、消失的时间,同时观察术后并发症发生情况。结果与结论:62例患者均获得12个月以上随访。复合骨髓干细胞植骨组移植骨界限模糊时间和消失时间均短于单纯植骨组(P<0.05)。复合骨髓干细胞植骨组1例出现排异反应,使用免疫抑制剂治疗2周后痊愈,两组病例均未出现感染。结果表明同种异体骨复合自体骨髓干细胞植骨能明显促进骨融合和骨缺损的愈合。