Kinetics of engraftment in patients with hematologic malignancies given allogeneic hematopoietic cell transplantation after nonmyeloablative conditioning

We analyzed the kinetics of donor engraftment among various peripheral blood cell subpopulations and their relationship to outcomes among 120 patients with hematologic malignancies given hematopoietic cell transplantation (HCT) after nonmyeloablative conditioning consisting of 2 Gy total body irradiation (TBI) with or without added fludarabine. While patients rapidly developed high degrees of donor engraftment, most remained mixed donor/host chimeras for up to 180 days after HCT. Patients given preceding chemotherapies and those given granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cell (G-PBMC) grafts had the highest degrees of donor chimerism. Low donor T-cell (P = .003) and natural killer (NK) cell (P = .004) chimerism levels on day 14 were associated with increased probabilities of graft rejection. High T-cell chimerism on day 28 was associated with an increased probability of acute graft-versus-host disease (GVHD) (P = .02). Of 93 patients with measurable malignant disease at transplantation, 41 achieved complete remissions a median of 199 days after HCT; 19 of the 41 were mixed T-cell chimeras when complete remissions were achieved. Earlier establishment of donor NK-cell chimerism was associated with improved progression-free survival (P = .02). Measuring the levels of peripheral blood cell subset donor chimerisms provided useful information on HCT outcomes and might allow early therapeutic interventions to prevent graft rejection or disease progression.     

异基因造血干细胞移植后早期嵌合状态研究

目的:建立多重PCR扩增短串联重复序列(STR)结合毛细管电泳,定量检测异基因造血干细胞移植(Allo-HSCT)后受者体内供、受体来源血细胞嵌合率的方法及灵敏度。并探讨该方法的早期定量检测对Allo-HSCT后白血病早期复发进行干预的指导意义。方法:将2名健康成人静脉血分离单个核细胞,并按供、受体单个核细胞不同百分比制成模拟嵌合体标本并提取基因组DNA,用多重PCR扩增后,进行毛细管电泳,确定基因位点及相应峰面积。15例Allo-HSCT患者移植前采集供、受者外周血,移植后采集受者外周血,分别提取DNA,进行PCR扩增和毛细管电泳,确定STR基因位点及峰面积,计算混合样本中2单个样本DNA含量百分比及Al-lo-HSCT患者嵌合率。结果:①当混合样本中某一细胞比例为4%以上时,即可从电泳图中明确区分混合样本中该单个样本的基因型,且扩增前细胞百分率与扩增后两者峰面积比值呈直线相关;②所有患者在移植后2周内均出现供者来源的细胞,13例患者在术后30d内均达稳定嵌合。2例患者移植后出现不稳定混合嵌合状态或复发,经干预治疗后转为完全嵌合状态。结论:STR毛细管电泳PCR方法非常敏感,能应用于Allo-HSCT后早期嵌合体的定量检测。嵌合状态的早期定量检测对了解Allo-HSCT后的移植物的植入状态和指导早期干预治疗均有重要参考价值。     

异基因造血干细胞移植治疗恶性血液病(附9例报告)

9例恶性血液病患者 ,采用异基因外周血干细胞移植 ( allo- HSCT)治疗 7例 ,外周血与骨髓干细胞混合移植 2例 ;供者均为人白细胞相关抗原 ( HL A)完全相合同胞兄妹。预处理方案用马利兰 ( BU)、环磷酰胺( CY) ,环孢霉素 A ( Cs A)联合短程甲氨蝶呤 ( MTX)预防移植物抗宿主病 ( GVHD)。结果 :患者治疗后均重建造血 ,中性粒细胞≥ 0 .5× 10 9/ L 的中位数为 14天 ,血小板≥ 2 0× 10 9/ L 的中位数为 16天。发生急性 GVHD5例 ,慢性 GVHD2例 ,肝静脉闭塞病 ( VOD) 1例 ,巨细胞病毒血症 9例。2例 ABO血型不合者移植后未发生溶血及纯红再障。1例耐药复发淋巴瘤 ,供、受者均为乙肝病毒携带者 ,移植后达完全缓解。中位随访时间 15个月 ,无病生存 8例。认为 allo- HSCT是治疗恶性血液病 (尤其是耐药复发者 )的有效方法 ,ABO血型不合及乙肝供者不是移植的障碍     

Impact of hepatitis C virus seropositivity on survival after allogeneic hematopoietic stem cell transplantation for hematologic malignancies

Background

Because hepatitis C virus infection causes hepatic and immunological dysfunction, we hypothesized that seropositivity for this virus could be associated with increased non-relapse mortality after allogeneic hematopoietic stem cell transplantation.

Design and Methods

We performed a case-control study of the outcomes of patients who were hepatitis C virus seropositive at the time of allogeneic hematopoietic stem cell transplantation (N=31). Patients positive for hepatitis C virus were considered candidates for stem cell transplantation only if they had no significant evidence of hepatic dysfunction. Matched controls (N=31) were seronegative for viral hepatitides and were paired according to age, diagnosis, disease stage, conditioning regimen and donor type. We also compared the hepatitis C virus seropositive patients to all seronegative patients (all controls, N=1800) transplanted during the same period, to adjust for other confounding effects.

Results

The median age of the seropositive patients was 49 (range 26–72); 15 had acute myeloid leukemia/myelodysplastic syndrome, 6 had chronic myeloid leukemia/myeloproliferative disease, 6 non-Hodgkin’s lymphoma, 2 myeloma, 1 acute lymphocytic leukemia and 1 Hodgkin’s lymphoma; 61% had poor risk disease; 68% had related donors; 68% received reduced intensity conditioning; 7 patients had mildly abnormal alanine transaminase levels (all less than three times the upper limit of normal) and 1 patient had minimally elevated bilirubin. These characteristics were similar to those of the matched control group. Median overall survival was 3, 18 and 20 months, and 1-year survival was 29%, 56% and 56%, in the hepatitis C virus, matched and all controls groups, respectively (hazard ratio for death 3.1, 95% confidence interval 1.9–5.6, p<0.001 in multivariate analysis). Non-relapse mortality at 1 year was 43%, 24% and 23%, respectively (hazard ratio 3.3, 95% confidence interval 1.8–7.1, p<0.01). Disease progression and graft-versus-host disease rates were comparable.

Conclusions

Hepatitis C virus seropositivity is a significant risk factor for non-relapse mortality after allogeneic hematopoietic stem cell transplantation even in patients with normal or minimally abnormal liver function tests.     

Efficacy and safety of Venetoclax in the treatment of 25 patients with recurrent hematologic malignancies after an allogeneic hematopoietic stem cell transplantation

<正>Objective To investigate the efficacy and safety of preemptive/salvage therapy with venetoclax(VEN) in patients with recurrence after allogeneic hematopoietic stem cell transplantation(allo-HSCT).     

Genetic variations in T-cell activation and effector pathways modulate alloimmune responses after allogeneic hematopoietic stem cell transplantation in patients with hematologic malignancies

Background

Recently, several important polymorphisms have been identified in T-cell activation and effector pathway genes and have been reported to be associated with inter-patient variability in alloimmune responses. The present study was designed to assess the impact of these genetic variations on the outcomes of allogeneic hematopoietic stem cell transplantation.

Design and Methods

We first investigated ten single nucleotide polymorphisms in six genes, CD28, inducible co-stimulator, cytotoxic T-lymphocyte antigen 4, granzyme B, Fas and Fas ligand, in 138 pairs of patients and their unrelated donors and a second cohort of 102 pairs of patients and their HLA-identical sibling donors.

Results

We observed that patients receiving stem cells from a donor with the cytotoxic T-lymphocyte antigen 4 gene CT60 variant allele (AA genotype) had a reduced incidence of grades II–IV acute graft-versus-host disease; however, they experienced early cytomegalovirus infection and relapsed more frequently, which suggested an interaction between the donor cytotoxic T-lymphocyte antigen 4 gene CT60 AA genotype and reduced T-cell alloreactivity. Furthermore, an unrelated donor with the granzyme B +55 variant genotype (AA) was an independent risk factor for development of grades II–IV acute graft-versus-host disease (P=0.024, RR=1.811). Among patients with acute myelogenous leukemia, those with the Fas -670 TT genotype were at higher risk of relapse (P=0.003, RR=3.823). The presence of these susceptible alleles in the donor and/or patient resulted in worse overall survival (54.9% versus 69.5%, P=0.029).

Conclusions

Our data suggest that genotype analysis of T-cell activation and effector pathway genes can be used for risk assessment for patients with hematologic malignancies before hematopoietic stem cell transplantation.Key words: CD28, inducible co-stimulator, cytotoxic T-lymphocyte antigen 4, granzyme B, Fas, Fas ligand, polymorphism, hematopoietic stem cell transplantation     

Strategies and clinical implications of chimerism diagnostics after allogeneic hematopoietic stem cell transplantation

Analysis of donor chimerism has become a routine method for the documentation of engraftment after allogeneic hematopoietic stem cell transplantation (HSCT). In recent years several groups have also focused on the application of this technique for the detection of relapsing disease after allogeneic HSCT. This review addresses technical issues (sensitivity, specificity) and discusses the advantages and limitations of methods currently used for chimerism analysis and their usefulness for the detection of MRD. In addition, the potential impact of novel procedures, e.g. subset chimerism or real-time PCR-based procedures, is discussed.     

Monthly prospective analysis of hematopoietic chimerism after allogeneic hematopoietic cell transplantation

Summary:Hematopoietic chimerism (HpC) was assayed monthly using a sensitive, polymerase chain reaction (PCR) -based method in consecutive patients. Between January 1998 and April 2002, 181 patients underwent non-T cell depleted allogeneic hematopoietic cell transplantation (HCT). A total of 163 patients were evaluable for HpC at 1 month (11 early deaths; no informative band for HpC analysis/no genomic DNA in seven). In all, 53 of 163 patients (33%, median recipient DNA of 15% (range 5-95)), 39 of 151 patients (26%), and 27 of 142 patients (19%) showed mixed chimerism (MC) at 1, 2, and 3 months after HCT, respectively. Conditioning regimen (busulfan-fludarabine-ATG vs BuCy, relative risk 3.99 (95% CI 1.16-10.92)), neutrophil engraftment (>/=day 17 vs /=5% recipient DNA at 1 month. Five patients experienced secondary graft failure. All five patients showed MC at 1 month with median recipient DNA of 40%. None of the 109 patients with complete chimerism experienced graft failure (P=0.002). Our study showed that MC shown on monthly analysis of HpC after allogeneic HCT is a significant predictor of secondary graft failure.Bone Marrow Transplantation (2003) 32, 423-431. doi:10.1038/sj.bmt.1704147     

Liver graft-versus-host disease after donor lymphocyte infusion for relapses of hematologic malignancies post allogeneic hematopoietic stem cell transplantation

Graft-versus-host disease (GVHD) is the commonest complication after donor lymphocyte infusion (DLI). In 19 patients undergoing DLI for relapses of hematologic malignancies post hematopoietic stem cell transplantation (HSCT), 11 developed GVHD, of whom nine had isolated liver involvement, and two had liver and skin involvement. The clinical diagnosis of liver GVHD was hepatitic in six patients (55%) and classical in five patients (45%). Patients with GVHD post-DLI showed a different clinical pattern when compared to a cohort of 106 cases of GVHD post-HSCT, in having significantly more isolated liver involvement (9/11 vs 17/106, P<0.001), and less skin (2/11 vs 80/106, P<0.001) and gut (0/11 vs 28/106, P<0.001) involvement. However, liver GVHD post-DLI and post-HSCT had comparable patient characteristics, underlying diseases, clinical subtypes (classical and hepatitic) and response to treatment.     

Differential epitope mapping of antibodies to PDC-E2 in patients with hematologic malignancies after allogeneic hematopoietic stem cell transplantation and primary biliary cirrhosis

A unique characteristic of the autoimmune liver disease primary biliary cirrhosis (PBC) is the presence of high-titer and extremely specific autoantibodies to the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Autoantibodies to PDC-E2 antigen have only been detected in patients with disease or in those who subsequently develop PBC. One exception has been a subgroup of patients with multiple myeloma (MM) who underwent allogeneic hematopoietic stem cell transplantation (HSCT) and received donor lymphocyte infusions (DLIs) after transplantation. These patients developed high-titer antibodies to a variety of myeloma-associated antigens, including PDC-E2, coincident with rejection of myeloma cells in vivo. To examine the specificity of autoantibodies to PDC in these patients, we screened sera from patients with MM, chronic leukemias, monoclonal gammopathy of unknown significance (MGUS), PBC, and healthy donors. Three of 11 patients with MM (27%) and 2 of 6 patients with chronic leukemias (33%) developed anti-PDC-E2 antibodies in association with DLI response; 2 of 12 (17%) patients in the MGUS pretreatment control population also had detectable anti-PDC responses. Interestingly, the epitope specificity of these PDC-E2 autoantibodies was distinctive, suggesting that the mechanisms leading to loss of tolerance in the transplantation patients are distinct from PBC.     

Human pulmonary chimerism after hematopoietic stem cell transplantation

Many of the body's tissues once thought to be only locally regenerative may, in fact, be actively replaced by circulating stem cells after hematopoietic stem cell transplantation. Localization of donor-derived cells ("chimerism") has recently been shown to occur in the lungs of mice after either hematopoietic stem cell transplantation or infusion of cultured marrow. To determine whether tissues of the human lung might be similarly derived from extrapulmonary sources, we examined lung specimens from a retrospective cohort of female allogeneic hematopoietic stem cell transplant recipients who received stem cells from male donors. Tissue samples from three such patients who had undergone diagnostic lung biopsy or autopsy were examined. Slides were stained by immunohistochemistry for cytokeratin (epithelium) and platelet endothelial cell adhesion molecule, CD31 (PECAM) (endothelium) and were imaged and then examined by fluorescent in situ hybridization analysis to identify male cells. The resulting overlapping in situ hybridization and immunohistochemistry images were examined for the presence and, if present, cell type of donor cells in the lung. We found significant rates of epithelial (2.5-8.0%) and endothelial (37.5-42.3%) chimerism. These results suggest that significant chimerism of the human lung may follow hematopoietic stem cell transplantation and that adult human stem cells could potentially play a therapeutic role in treatment of the damaged lung.     

四核苷酸重复序列估价异基因外周血干细胞移植后嵌合状态

目的 确定适合于检测中国人群异基因外周血干细胞移植后嵌合状态的5个四核苷酸重复序列（或称位点）,探讨嵌合状态与预后的关系。方法 PCR扩增140份脐带血和17对移植供受者基因组DNA,电泳,银染,据所得片段大小考察5个位点在人群中的多态性及移植后嵌合状态,结合临床分析。结果 位点多态性：各位点发现7个以上等位基因（7－12）,16种以上基因型（16－28）,杂合度仅CSF1PO低于70％。移植患者中完全嵌合状态（CC)12例,1例死于移植物抗宿主病;混合嵌合状态（MC）1例死亡;受者型1例死亡;MC经二次移植转为CC1例无复发。2例嵌合情况不同。结论 上述5位点多态性较强,可成功检测大多数嵌合状态。移植患者随着体内受者细胞成分的增加,预后渐差。     

Mixed hematopoietic chimerism following bone marrow transplantation for hematologic malignancies

Twenty-nine of 172 patients (17%) who received an allogeneic bone marrow transplant (BMT) from histocompatible sibling donors for hematologic malignancies were mixed hematopoietic chimeras; ie, they had a mixture of donor and host hematopoietic or lymphohematopoietic cells at greater than or equal to 14 days after transplantation. Twenty- four of the 29 mixed chimeras (83%) have remained in continuous complete remission for up to 116 months (greater than 9 years) following BMT. Four of the 29 patients (14%) have had recurrent leukemia, and 7 of the 29 (24%) have had moderate or severe graft-v- host disease (GVHD). Twelve of these 29 patients have persisted as stable mixed chimeras for greater than or equal to 2 years after BMT, whereas other patients converted to all donor-type hematopoiesis. The incidence of mixed chimerism was independent of the pretransplant regimen, the donor or recipient age (less than 20 v greater than 20 years), remission status (first complete remission of acute leukemia and first chronic phase of chronic myelocytic leukemia v later stages of disease), and type of leukemia. Our data indicate that mixed hematopoietic chimerism is not rare after BMT for hematologic malignancies and that its presence is compatible with long-term disease- free survival. Prospective studies of mixed chimerism after BMT are warranted to achieve better understanding of its biologic importance.     

不同供者来源异基因造血干细胞移植治疗恶性血液病69例疗效和预后分析

目的评价和比较无关供者与同胞供者异基因造血干细胞移植治疗恶性血液病的临床疗效。方法回顾性分析东南大学附属中大医院血液科2006年4月至2011年12月进行异基因造血干细胞移植的69例恶性血液病患者的临床资料,根据造血干细胞(HSC)来源的不同,将患者分为无关供者(URD组)30例和同胞供者(RD组)39例。对两组患者移植后中性粒细胞和血小板的植入时间、细胞嵌合状态、移植物抗宿主病(GVHD)、感染的发生率以及复发和总生存(OS)率等长期随访结果进行分析。结果 69例达到完全稳定的供者植入,URD组和RD组中性粒细胞和血小板植活的中位时间分别为11 d、10 d和15 d、14 d(均P>0.05)。共33例发生急性GVHD,其中URD组发生I～Ⅱ度GVHD 12例(40.0%);7例(23.3%)发生Ⅲ～Ⅳ度GVHD;RD组12例(30.8%)发生Ⅰ～Ⅱ度GVHD,与URD组相比差异无统计学意义(P=0.455);2例(5.1%)发生Ⅲ～Ⅳ度GVHD,与URD组相比差异有统计学意义(P=0.035)。URD和RD组的5年OS率分别为56.7%和64.1%(P=0.621)。结论无关供者和同胞供者异基因造血干细胞移植均可成功治疗恶性血液病,疗效相似;但与RD比较,URD的Ⅲ～Ⅳ度aGVHD的累积发生率高、早期感染率较高、而复发率较低。     

Analysis of efficacy and prognosis of allogeneic hematopoietic stem cell transplantation from different donors in treatment of hematologic malignancies

Objective To investigate the clinical efficacy of allogeneic hematopoietic stem cell transplantation(allo-HSCT) from unrelated donors and that from related donors in treatment of hematologic malignancies. Methods     

二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性

目的 探讨应用二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性.方法 回顾性分析1999年10月-2010年3月在北京大学血液病研究所接受二次造血干细胞移植作为挽救治疗的25例移植后复发患者的资料.除1例移植前未达缓解状态的患者外,其余24例患者在第1次移植后的中位8.8(1～55)个月出现复发,经过中位3(0.3～20)个月的治疗,接受了二次移植.2次移植中位间隔10.6(0.6～59.0)个月.结果 25例患者预处理方案主要为含全身放疗(TBI)或改良的BUCY(马利兰+环磷酰胺)方案.全部患者达到白细胞植活标准,并生存超过30 d,有3例患者发生肝窦阻塞综合征,放射性皮炎以及急性心肌梗死等严重预处理相关毒性,但经治疗后均恢复.截至2011年1月,经过中位9.1(2.0～131.1)个月的随访,8例患者生存,总的生存率为30.9%.12例患者复发,复发率55.7%,复发中位时间是二次移植后的4.4(1.0～8.5)个月.7例患者因移植相关合并症死亡,非复发病死率35.1%.单因素分析发现患者二次移植时是否缓解与生存期有显著关联(P=0.009).结论 适度降低强度的预处理方案可以使二次异基因造血干细胞移植安全地应用于移植后复发的患者,保证移植物的植入,非复发病死率亦在可接受范围内.     

二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性

目的 探讨应用二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性.方法 回顾性分析1999年10月-2010年3月在北京大学血液病研究所接受二次造血干细胞移植作为挽救治疗的25例移植后复发患者的资料.除1例移植前未达缓解状态的患者外,其余24例患者在第1次移植后的中位8.8(1～55)个月出现复发,经过中位3(0.3～20)个月的治疗,接受了二次移植.2次移植中位间隔10.6(0.6～59.0)个月.结果 25例患者预处理方案主要为含全身放疗(TBI)或改良的BUCY(马利兰+环磷酰胺)方案.全部患者达到白细胞植活标准,并生存超过30 d,有3例患者发生肝窦阻塞综合征,放射性皮炎以及急性心肌梗死等严重预处理相关毒性,但经治疗后均恢复.截至2011年1月,经过中位9.1(2.0～131.1)个月的随访,8例患者生存,总的生存率为30.9%.12例患者复发,复发率55.7%,复发中位时间是二次移植后的4.4(1.0～8.5)个月.7例患者因移植相关合并症死亡,非复发病死率35.1%.单因素分析发现患者二次移植时是否缓解与生存期有显著关联(P=0.009).结论 适度降低强度的预处理方案可以使二次异基因造血干细胞移植安全地应用于移植后复发的患者,保证移植物的植入,非复发病死率亦在可接受范围内.     

二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性

目的 探讨应用二次异基因造血干细胞移植治疗移植后复发患者的疗效和安全性.方法 回顾性分析1999年10月-2010年3月在北京大学血液病研究所接受二次造血干细胞移植作为挽救治疗的25例移植后复发患者的资料.除1例移植前未达缓解状态的患者外,其余24例患者在第1次移植后的中位8.8(1～55)个月出现复发,经过中位3(0.3～20)个月的治疗,接受了二次移植.2次移植中位间隔10.6(0.6～59.0)个月.结果 25例患者预处理方案主要为含全身放疗(TBI)或改良的BUCY(马利兰+环磷酰胺)方案.全部患者达到白细胞植活标准,并生存超过30 d,有3例患者发生肝窦阻塞综合征,放射性皮炎以及急性心肌梗死等严重预处理相关毒性,但经治疗后均恢复.截至2011年1月,经过中位9.1(2.0～131.1)个月的随访,8例患者生存,总的生存率为30.9%.12例患者复发,复发率55.7%,复发中位时间是二次移植后的4.4(1.0～8.5)个月.7例患者因移植相关合并症死亡,非复发病死率35.1%.单因素分析发现患者二次移植时是否缓解与生存期有显著关联(P=0.009).结论 适度降低强度的预处理方案可以使二次异基因造血干细胞移植安全地应用于移植后复发的患者,保证移植物的植入,非复发病死率亦在可接受范围内.