Czekalska Sylwia, Sacha Tomasz, Piatkowska-Jakubas Beata, Zawada Magdalena, Florek Izabela, Cwynar Dorota, Skotnicki Aleksander B
Katedra i Klinika Hematologii CMUJ Kraków.
Przegl Lek. 2010;67(12):1282-91.
Allogeneic hematopoietic stem cell transplantation (alloSCT) is a curative treatment for many patients suffering from malignant and non-malignant hematological disorders. Successful transplantation is a process that requires the engraftment of transplanted pluripotent hematopoietic stem cells which re-establish normal hematological and immunological systems. Distinguishing between host and donor origin of bone marrow and blood cells is vitally important for monitoring of the engraftment process. One of the most useful tools for engraftment monitoring is the assessment of hematopoietic chimerism. Which occurs after alloHSCT and describes the percentage of donor hematopoietic and lymphoid cells in a transplant recipient. 38 adult patients, after alloSCT performed in Katedra i Klinika Hematologii Collegium Medicum UJ entered the study and the total number of transplantations was 43. The evaluation of hematopoietic chimerism was based on PCR amplification of polymorphic non-coding DNA sequences--short tandem repeats (STR-PCR). The main tool was a semiquantitative method--fragment length analysis. The product of amplification was analyzed using the sequencer. The second method was based on a quantitative Real Time PCR technique (RQ-PCR) based on SYBRgreen chemistry. There were performed amplification of biallelic non-coding DNA sequences with short insertions or deletions. Hematopoietic chimerism evaluations were performed on +30, +60, +90, +120, +150, +180, +270 and +360 day and then every 6 months post alloSCT on peripheral blood or bone marrow samples. STR-PCR and RQ-PCR chimerism assays were compared and results evidenced the greater sensitivity of RQ-PCR method. There were not crucial differences in the results of chimerism evaluation obtained by means of these two methods. The analysis of chimerism kinetics after allogeneic stem cell transplantation allowed to modify the post-transplantation-treatment in 3 patients after alloNMSCT leading to increase of donor-origin hematopoiesis in transplant recipients (in 2 pts decision of DLI, 1 of them withdrawal of immunosuppression, 1 pt giving G-CSF). The results of chimerism monitoring confirmed that the failure of achieving a CC or lost of CC can predict the relapse of the disease.
异基因造血干细胞移植(alloSCT)是许多患有恶性和非恶性血液系统疾病患者的一种治愈性治疗方法。成功的移植是一个需要移植的多能造血干细胞植入并重新建立正常血液和免疫系统的过程。区分骨髓和血细胞的宿主来源与供体来源对于监测植入过程至关重要。植入监测最有用的工具之一是评估造血嵌合体。造血嵌合体发生在alloHSCT之后,描述了移植受者中供体造血和淋巴细胞的百分比。38名成年患者在雅盖隆大学医学院血液学系和诊所接受alloSCT后进入研究,移植总数为43例。造血嵌合体的评估基于多态性非编码DNA序列——短串联重复序列(STR-PCR)的PCR扩增。主要工具是半定量方法——片段长度分析。使用测序仪分析扩增产物。第二种方法基于基于SYBRgreen化学的定量实时PCR技术(RQ-PCR)。对具有短插入或缺失的双等位基因非编码DNA序列进行扩增。在alloSCT后的第30、60、90、120、150、180、270和360天,然后在alloSCT后每6个月对外周血或骨髓样本进行造血嵌合体评估。比较了STR-PCR和RQ-PCR嵌合体检测方法,结果证明RQ-PCR方法具有更高的灵敏度。通过这两种方法获得的嵌合体评估结果没有关键差异。对异基因干细胞移植后嵌合体动力学的分析使得3例alloNMSCT患者在移植后调整治疗方案,从而增加了移植受者中供体来源的造血(2例决定进行供体淋巴细胞输注,其中1例停用免疫抑制,1例给予粒细胞集落刺激因子)。嵌合体监测结果证实,未能实现完全嵌合(CC)或失去CC可预测疾病复发。