异基因造血干细胞移植患者外周血TCR Vβ亚家族克隆性增殖动态变.doc

异基因造血干细胞移植患者外周血TCR Vβ亚家族克隆性增殖动态变 　 作者：贾新颜,王健民,李扬秋,陈少华,章卫平,陈莉 【摘要】 本研究观察异基因造血干细胞移植(alloHSCT)患者外周血T 细胞受体（TCR） Vβ亚家族克隆性增殖的动态变化，分析T细胞的克隆性演变与GVHD的关系。利用RTPCR方法扩增70例次alloHSCT后患者（其中17例次出现GVHD）外周血单个核细胞中24个TCR Vβ基因的互补决定区3（CDR3），PCR产物经荧光标记和基因扫描分析CDR3长度, 确定T细胞的克隆性。结果表明：移植患者T细胞增殖一般都经历由单克隆向多克隆演变的过程，在移植后60-90天时，逐渐由单克隆转为单克隆和多克隆表达大致各占一半。120天以后，无GVHD患者大多转为多克隆性表达；而GVHD患者受免疫抑制剂和GVHD的影响，直到1年以上仍有部分呈单克隆表达趋势。GVHD患者发生GVHD或靶器官受累最明显的时候，外周血TCR Vβ亚家族的表达主要呈现单/双克隆的表现，而经过免疫抑制治疗病情好转后，部分患者出现由寡克隆表达转为多克隆增殖趋势。结论：移植后早期患者尤其是合并GVHD的患者，T细胞呈克隆性增殖和T细胞受体的倾向性利用；随着造血和免疫的恢复，TCR Vβ亚家族表达重新恢复趋向正常的多克隆性演变。 【关键词】 造血干细胞移植 Clonal Kinetic Proliferative Change of TCRVβ Subfamilies in Peripheral Blood of Patients Transplanted with Allogeneic Hematopoietic Stem Cells and Its Relation to GVHD Abstract This study was purposed to investigate the dynamic change of clonal proliferation of T cell receptor V subfamilies in peripheral blood of patients received allohematopoietic stem cell transplantation (alloHSCT) and to analyze the relationship between T cell clonal proliferative changes and GVHD. The peripheral blood mononuclear cell samples from 70 cases (17 GVHD patients) undergoing alloPBCST patients were detected for CDR3 (complementarity determining region 3 repertoire analysis of T cell receptor Vβgene) using reversetranscriptasepolymerase chain reaction (RTPCR)．The products were further analyzed by genescan to identify T cell clonality．The sesults showed that the patients of HSCT generally passed through a transformation from monoclone to polyclone. At day 60-90 after HSCT, half of the cases were monoclonal and the remainders were polyclonal. After 120 days, most of patients without GVHD transferred into polyclones, however, patients with GVHD remained monoclonal after one year because of immunosuppressive agents and GVHD itself. The peripheral blood of GVHD patients mainly expressed monoclone/biclone at the time of target organ damage conspicuously, after medication intervention, partial monoclone or bioclone