荧光定量PCR检测T细胞受体删除环在亲缘与非亲缘异基因造血干细胞移植后免疫重建早期的应用

Quantitative analysis of T-cell receptor excision cycles in patients after related or unrelated donor hemopoietic stem cell transplantation in early immune reconstitution period

目的检测接受亲缘与非亲缘的异基因造血干细胞移植(allogeneic hemopoietic stem cell transplant,alloHSCT)的恶性血液病患者免疫重建早期T细胞受体删除环(signal joint T-cell receptor excision circle,sjTREC)的水平,分析供受者间亲缘关系对免疫重建早期的影响。方法采用TaqMan探针,Real-Time PCR技术,对6例(2例亲缘,4例非亲缘)allo-HSCT恶性血液病患者在移植前及移植后6个月的外周血sjTREC进行绝对定量检测。结果每1 000个细胞中各组sjTREC水平分别为:亲缘组移植前(17.05±22.28)拷贝,移植后(24.26±25.50)拷贝;非亲缘组移植前(6.74±12.85)拷贝,移植后(28.30±19.12)拷贝。结论两组患者移植后的sjTREC水平较移植前均有所上升,但无论移植前后两组之间的差异均不明显,提示在allo-HSCT患者移植后的免疫重建受诸多因素共同影响,供者是否具有亲缘关系在小样本检测中未发现明显差异。

Objective The purpose of the study was to compare recipients with related donor hemopoietic stem cell trans- plantation to unrelated ones by investigating the copies of sjTREC in peripheral blood. Methods The sjTREC of recipients （re- lated donor group=2; unrelated donor group= 4） was detected by using absolute quantitative Real-Time PCR with TaqMan probes. Results According to standard plasmid concentration gradients, in 1 000 cells, the pretransplant sjTREC were （17. 05± 22.28） copies （related donor group） and （6.74± 12.85） copies （unrelated donor group）; after transplantation, which were （24. 26±25.50） copies （related donor group） and （28.30±19.12） copies （unrelated donor group）. Conclusion The difference between related donor group and unrelated group ever pretransplant or after is not significant. It indicates many factors impact the function of thymus are very complex, and the relationship of donor is one of them, but it does not obviously controlled the thymus in early immune reconstitution period.