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外周造血干细胞移植后及出现GVHD时T细胞β链CDR3谱型分析
引用本文:姚新生,周明乾,马骊,罗微,冯晓勤,温茜,王小宁.外周造血干细胞移植后及出现GVHD时T细胞β链CDR3谱型分析[J].南方医科大学学报,2008,28(4):529-533.
作者姓名:姚新生  周明乾  马骊  罗微  冯晓勤  温茜  王小宁
作者单位:1. 南方医科大学生物技术学院分子免疫研究所,广东,广州,510515
2. 南方医院儿科移植中心,广东,广州,510515
3. 华南理工大学生物科学与工程学院,广东广州,510641
基金项目:教育部跨世纪优秀人才培养计划 , 国家自然科学基金 , 广东省自然科学基金
摘    要:目的 初步探讨外周造血干细胞(PBSC)移植后以及出现移植物抗宿主疾病(GVHD)时,外周血T细胞a链的CDR3谱型变化和特征.方法 采用免疫扫描谱型分析技术,监测1例重型β-地中海贫血移植前、移植后23 d、GVHD时(移植后28 d)患者外周血单核细胞(PBMC)及供者PBSC标本T细胞a链的CDR3谱型变化,基因扫描(GeneScan)和测序分析克隆性增生T细胞TCR分子特征.结果 患者移植前PBMC的24个TCR BV家族CDR3谱型均呈高斯分布,部分家族在移植后23 d、和GVHD时呈现为寡峰和单峰,同时部分家族消失,单克隆增生T细胞TCR beta链的测序结果 显示不同的CDR3区组成,设计特异的CDR3引物进一步分析提示其可能来源于干细胞移植后的分化.结论 PBSC移植后23 d,PBMC中的多数24TCRBV家族CDR3谱型表现为多克隆,GVHD时,部分家族出现明显的单、寡克隆增生,这些特异T细胞可能在GVHD中发挥重要作用.对移植前后TCRCDR3谱型和特异TCR分子特征的分析将有助于PBSC移植后免疫重建的评估和出现排斥反应时的特异性治疗.

关 键 词:T淋巴细胞受体  互补决定区3  免疫扫描谱型分析  移植物抗宿主疾病  外周造血干细胞移植  GVHD  β链  谱型  分析  patient  T  cells  peripheral  blood  expansion  stem  cell  transplantation  特异性治疗  反应时  评估  免疫重建  作用  寡克隆  多克隆  表现  来源  设计
文章编号:1673-4254(2008)04-0529-04
修稿时间:2007年11月18

CDR3 spectratyping and clonal expansion of peripheral blood T cells in a patient with β-mediterranean anaemia following peripheral blood stem cell transplantation
YAO Xin-sheng,ZHOU Ming-qian,MA Li,LUO Wei,Feng Xiao-qin,WEN Qian,WANG Xiao-ning.CDR3 spectratyping and clonal expansion of peripheral blood T cells in a patient with β-mediterranean anaemia following peripheral blood stem cell transplantation[J].Journal of Southern Medical University,2008,28(4):529-533.
Authors:YAO Xin-sheng  ZHOU Ming-qian  MA Li  LUO Wei  Feng Xiao-qin  WEN Qian  WANG Xiao-ning
Institution:Institute of Molecular Immunology, School of Biotechnology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Abstract:OBJECTIVE: To study the CDR3 spectratyping and clonal expansion of T cells in the peripheral blood mononuclear cells (PBMCs) of patients with beta-mediterranean anemia patient undergoing allogeneic peripheral blood stem cell (PBSC) transplantation. METHODS: The total RNA was isolated from the PBMCs of a nine-year-old boy with beta-mediterranean anemia before and after PBSC transplantation, and at the time of occurrence of graft-versus-host disease (GVHD). The CDR3 length was analyzed using immunoscope technique, and the characteristics of the T cell receptor (TCR) on the T cells with clonal expansion were examined by gene sequencing. RESULTS: The 24 TCR BV CDR3 repertoire showed Gaussian distribution in the PBMCs isolated before the transplantation, and some of the TCR BV family CDR3 showed skewing in the PBMCs isolated 23 days after transplantation and at the onset of GVHD (28 days after transplantation), suggesting the clonal expansion of the donor PBSCs. CONCLUSION: The host PBMCs show muti-clonal expansion 23 days after PBSC transplantation, and in the stage of GVHD, some of the TCR BV family T cells show significant monoclonal expansion. Analysis of TCR CDR3 spectratyping and the molecular characteristics of specific TCR may help evaluate the immune reconstitution following the transplantation and indicate specific treatment for potential GVHD.
Keywords:T cell receptors  complementarity determining region3  immunoscope spectratyping  graft versus host diseases  
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