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  

CDR3 spectratyping and clonal expansion of peripheral blood T cells in a patient with β-mediterranean anaemia following peripheral blood stem cell transplantation

作　　者：姚新生[1] 周明乾[1] 马骊[1] 罗微[1] 冯晓勤[2] 温茜[1] 王小宁[3] 

机构地区：[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

出　　处：《南方医科大学学报》2008年第4期529-532,536,共5页Journal of Southern Medical University

基　　金：教育部新世纪优秀人才支持计划(NCET-07-0410);国家自然科学基金(30771952);广东省自然科学基金(07117783)~~

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!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23d、GVHD时(移植后28d)患者外周血单核细胞(PBMC)及供者PBSC标本T细胞β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23d、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23d,PBMC中的多数24TCR BV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TCR CDR3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。Objective To study the CDR3 spectratyping and clonal expansion ofT cells in the peripheral blood mononuclear cells （PBMCs） of patients with β-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 β-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. Conclusions 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.

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 

分 类 号：R392.4[医药卫生—免疫学]