混合脐血成人移植定量监测植入状态的实验研究  

Experimental Study on Quantitative Monitoring Engraftment of An Adult with Mixed Umbilical Cord Blood Transplantation

作　　者：邹红岩[1] 李桢[1] 邓志辉[1] 程良红[1] 吴国光[1] 

机构地区：[1]深圳市血液中心输血医学研究所,深圳518035

出　　处：《中国实验血液学杂志》2004年第2期179-184,共6页Journal of Experimental Hematology

摘　　要：为了定量监测和研究双份异基因脐血用于成人白血病患者移植后两份脐血的植入状态、嵌合体类型、供者细胞相对数量的动态变化及演变规律 ,采用荧光标记复合扩增短串联重复 (STR)位点嵌合体定量检测技术 ,对 1例急性髓细胞白血病成人患者移植两份 (脐血 1有核细胞数为 2 .5× 10 7/kg ,脐血 2有核细胞数为 1.5 3× 10 7/kg)HLA各 1个位点不相合的异基因脐血前后的序列血样进行 9个STR位点的检测 ;利用供、受者之间的差异位点定性判断脐血是否植入以及嵌合体类型 ;而后根据 377XLDNA测序仪上荧光扫描后两供者差异基因检出峰的峰面积计算脐血植入后患者体内两份脐血的细胞相对数量 ,定量分析供体细胞植入程度及演变规律 ;并与采用HLA差异基因对植入状态的分析结果进行对比。结果表明 :移植后 15天两份脐血同时植入 ,植入状态为完全双份供者嵌合体 ,患者体内脐血 1的相对细胞数量占 5 1.3% ,脐血 2占 4 8.7% ;30天时脐血 1嵌合体细胞上升为 70 .0 % ,脐血2嵌合体细胞下降为 30 .0 %。 5 2天时只检测到脐血 1的基因 ,植入状态转为完全单份供者嵌合体 ,有核细胞数少的一份脐血被排斥 ,有核细胞数多的一份长期植入。结论 :荧光标记复合扩增STR嵌合体定量检测可精确地描述两份脐血的植入程度及变化过程 。The purpose of this research was to monitor quantitatively and study the dynamic changes and development rules of engraftment, chimera types, as well as relative amount of donor cells after allogeneic transplantation of mixed umbilical cord blood from two units. An adult patient with acute myeloid leukemia received two units HLA one locus mismatched unrelated umbilical cord blood transplantation (2.5×10 7/kg karyocytes in umbilical cord blood unit 1, and 1 53×10 7/kg karyocytes in umbilical cord blood unit 2). Nine STR loci of the blood sample before and after transplantation were detetmined by quantitative detecting technique with fluorescence labeling polymerase chain reaction, while the engraftment and chimera types were qualitatively evaluated by comparing differential loci between the recipient and the donors. Then the relative proportion of chimera from two units of umbilical cord blood in the patient after transplantation was calculated according to the differential gene peak areas of two donors on 377XL DNA sequencer after fluorescence scanning, and the engraftment level and the development rules of donor cells were analyzed. In addition, the results were also compared with that of HLA loci distinct analysis for engraftment. The results showed that two umbilical cord blood units at 15 days after transplantation were engrafted simultaneously and revealed a complete chimerism of the two . The relative amounts of chimera from unit 1 vs that of unit 2 were 51.3% vs 48.7%; subsequently relative amounts of chimera from unit 1 went up to 70.0% at 30 days, and that from unit 2 declined to 30.0%. However, at 52 days, only the genotype of umbilical cord blood unit 1 was detected, so that the engraftment turned to a complete chimerism of a single donor type. The one with fewer karyocytes was rejected and the one with more karyocytes finally engrafted in long term. It is concluded that quantitatively detecting STR chimera with fluorescence labeling polymerase chain reaction can depict precisely the engra

关 键 词：白血病 脐血移植 混合脐血移植 嵌合体 短串联重复序列 

分 类 号：R331.1[医药卫生—人体生理学] R457.7[医药卫生—基础医学]