异基因外周血干细胞移植供受者嵌合体形成的动态检测  

Application of sequential monitoring of chimerism in HLA haploidentical family donor allogeneic peripheral blood stem cell transplantation

作　　者：曲建华[1] 温丙昭[1] 刘晓霞 董雷 哈力达.亚森[1] 江明[1] 陈瑢[1] 王蕾[1] 

机构地区：[1]新疆医科大学第一附属医院血液科,新疆乌鲁木齐830011 [2]新疆乌鲁木齐市刑侦支队,新疆乌鲁木齐830063

出　　处：《新疆医科大学学报》2009年第10期1430-1433,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区高技术研究与发展计划资助项目(200511109)

摘　　要：目的:建立荧光标记的多重扩增短串联重复序列(STR)结合毛细管电泳定量检测异基因外周血干细胞移植(allo-PBSCT)供受者嵌合体的方法,并探讨嵌合体形成与免疫耐受及临床预后的关系。方法:采用PE 9700扩增仪扩增15个STR位点:D8S1179、D21S11、D7S820、D3S1358、CSFIPO、THO1、D13S317、D16S539、D2S1338、D19S433、VWA、TPOX、D18S51、D5S818、FGA位点和1个性别位点Amelogenin,应用3130XL型DNA测序仪对35例异基因外周血干细胞移植患者上述位点进行基因型分析,并比较供受者移植前后的基因型,确定嵌合体的形成类型,以作为异基因造血干细胞移植中的植活证据,同时用无关个体的DNA混合实验进行可行性和准确性研究。结果:(1)其中32例在15个STR位点中均至少有两个可以区分供受者的有信息位点(有3例无移植前受者标本),35例患者均在移植后1个月时STR位点及性别位点转为供者型,形成了完全供者嵌合体(CC),且HLA单倍体相合的15例患者也达到了100%的供者植入,并在随访过程中持久不变。(2)扩增产物中供受者DNA含量的百分比同扩增前混合样本的比例呈显著直线相关(P<0.01),最小检出DNA百分比为5%。结论:(1)用基因分析仪对STR位点进行基因型检测具有快速、无污染和自动化高、可定量及灵敏度高等优点,用于检测allo-PBSCT植活状态是可靠的。(2)诱导免疫耐受的方法可以成功地诱导CC的形成。Objective： To establish multiple short tandem repeat （STR） amplification by fluorescence labe ling polymerase chain reaction （PCR） combined with capillary eleetrophoresis for quantitative determina- tion of chimerism, and to evaluate the relation between the type of chimerism and the outcome of allo- genenic peripheral blood stem cell transplantation （allo-PBSCT）. Methods： Thirty-five patients were ana- lyzed, including twenty with HLA identical sibling donors and fifteen with HLA haploidentical family do- nors, ABI PRISM 3130XL was used to genotype 15 microsatellite loci for D8Sl179, D2IS11, D7S820, D3S1358, CSFIPO, THO1, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA and sexual locus Amelogenin. The results of the donor and recipient in pre-transplantation and post- transplantation were analyzed to determine the type of chimerism. The feasibility and accuracy of this assay were tested in-vitro by using serial DNA mixtures from unrelated individuals. Results： （1） Discrimina- tion between donor and recipient was possible in all patients except for 3 patients whose pre-transplantation samples were not available, complete donor chimerism （CC） were detected in all patients at 1 month post- transplantation. （2） The result showed a clear correlation between the percentage of donor or recipient DNA 5%. ting, and the proportion of Conclusions.. （ 1 ） This and provide a reliable The measurement taken for mixed sample, with the limit of detection for a minor DNA percentage being approach allows a rapid, sensitive, automated and non-isotopic STR-PCR tes- alternative for assessment of the status of chimerism after allo-PBSCT. （2） the induction of immune tolerance in our study may induce CC successfully.

关 键 词：异基因外周血干细胞移植 移植物抗宿主病 嵌合体 

分 类 号：R457.7[医药卫生—治疗学]