非清髓性干细胞移植患者供体细胞嵌合率的动态定量研究  

Sequential and quantitative analysis of chimerism after non-myeloablative stem cell transplantation

作　　者：唐晓文[1] 吴德沛[1] 夏学鸣[1] 孙爱宁[1] 朱子玲[1] 阮长耿[1] 

机构地区：[1]苏州大学附属第一医院江苏省血液研究所,215006

出　　处：《中华内科杂志》2004年第2期98-101,共4页Chinese Journal of Internal Medicine

基　　金：江苏省 13 5重点医学人才基金资助项目 (RC2 0 0 2 0 3 3 ) ;江苏省卫生厅重点课题资助项目 (H2 0 13 )

摘　　要：目的　对非清髓性干细胞移植 (NST)后供受嵌合体形成动力学 ,嵌合体的转归 ,以及供体细胞嵌合率 (DC)对评价植入、复发、移植物抗宿主病 (GVHD)和长期生存的作用进行研究。方法　18例接受HLA全相合NST的患者进行DC的定量分析。术前采集供者和受者外周血 ,术后不同时间段采集受者外周血或骨髓。用QIAamp全血DNA抽提试剂盒提取样本DNA ,用AmpF/STRprofilerplus试剂盒进行 9个STR位点复合扩增后 ,产物用ABI 310遗传分析仪进行毛细管电泳 ,由Genescane和Genotype软件确定基因位点及峰面积 ,根据供受体基因型的差异计算供体细胞嵌合率。结果　 (1)移植早期植入表明 ,移植后 8d供体细胞开始占优势 (DC >6 0 % ) ,比造血重建提早 4d ;此外 ,由于移植前免疫状态不同 ,慢性髓细胞性白血病 (CML)患者供体细胞植入滞后于急性白血病和其他非恶性血液病患者 ;(2 )NST后嵌合状态有一个由混合嵌合 (MC)向完全供体细胞嵌合 (FDC)状态转化的过程 ;(3)FDC组GVHD的发生率高于MC组 (90 0 %、6 2 5 % ) ,而且从FDC状态的建立到急性GVHD发生的中位时间仅为 9d ;(4)移植后能获得无白血病生存的患者均有FDC或供体细胞高比例MC稳定嵌合的特点 ,而复发或排斥患者均在发生临床症状之前 ,出现供体细胞嵌合率的进行性下降。Objective To investigate the exact kinetics of donor chimerism (DC), outcome of mixed chimerism (MC) and prognostic role of chimerism in the evaluation of engraftment, disease relapse, GVHD and long term survival after nonmyeloablative stem cell transplantation (NST). Methods 18 patients who received HLA compatible NST were evaluated. Peripheral blood and bone marrow were collected before and after transplantation at different time. DNA was extracted using QIAmp blood mini kit. Nine different STR markers were co-amplified in a single reaction by commercial AmpF/STR profiler plus PCR amplification kit. Separation of the PCR products and fluorescence detection were performed with ABI prism 310 genetic analyzer with capillary electrophoresis. Genescan and genotype software were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on different allelic distribution types between the donor and recipient. Results (1) Serial STR-PCR analysis revealed that donor chimerism became dominant (DC>60%) by day 8; it preceded the detection of hematologic engraftment by an average of 4 days. It was also shown that chronic myeloid leukemia (CML) patients frequently had more delayed donor engraftment as compared with patients of acute leukemia or nonmalignant hematological diseases because the pretransplantation immune status of these two kinds of patients was different. (2) After NST, chimeric status had a process of conversion from the mixed chimerism (MC) to full donor chimerism (FDC). (3) The incidence of graft versus host disease (GVHD) of FDC group was higher than that of MC group (90.0% vs 62.5%).The average time between establishment of FDC and appearance of GVHD was 9 days. (4) Full donor chimerism and stable mixed chimerism with a high level of donor cells were compatible with disease free survival. On the contrary, progressive decrease of donor chimerism value was always followed by hematological relapse or graft rejection. Conclusions Sequencial and quantitati

关 键 词：非清髓性干细胞移植 供体细胞嵌合率 移植物抗宿主病 复发 定量检测 

分 类 号：R457[医药卫生—治疗学]