Fu Yue-wen, Wu De-pei, Sun Ai-ning, Feng Yu-feng, Chang Wei-rong, Zhu Zi-ling, Zhu Ping
Department of Hematology, The First Affiliated Hospital, Suzhou University, Suzhou 215006, China.
Zhonghua Xue Ye Xue Za Zhi. 2007 May;28(5):312-7.
To study the molecular characteristics of CDR3 repertoires of T cell receptor beta chain variable region (TCRBV) of T lymphocytic clones in leukemia recipients after allogeneic hematopoietic stem cell transplantation ( allo-HSCT).
RT-PCR was used to amplify 24 subfamily genes of TCRBV from peripheral blood (PB) lymphocytes in twenty-four leukemia patients underwent three kinds of allo-HSCT and in five normal donors as control. The PCR products were further analyzed by genescan to evaluate the clonality of BV subfamily and characteristics of CDR3 and calculate usage rate of BV subfamily. The monoclonal bands which associated with GVHD and CMV infection were obtained by denaturation polyacrylamide gel electrophoresis and sequenced. Comparison of the sequences of TCRBV CDR3 with other CDR3 sequences which associated with GVHD or CMV infection was reported.
2 approximately 19 months after transplantation, there were 6 approximately 14 BV subfamilies expressed and the polyclonal expression reached 33% in nine patients underwent haploidentical bone marrow transplantation(HI-BMT). In five patients underwent matched unrelated peripheral blood stem cell transplantation ( MU-PBSCT), there were 10 approximately 15 BV subfamilies expressed of which 45% were poly-clones. In 10 patients underwent matched sibling bone marrow transplantation(MS-BMT), 10 approximately 16 BV subfamilies were expressed and more than 48% of them were poly-clones. Monoclones and oligo-clones existed in 24 BV subfamilies but no common one monoclone BV subfamilies was found. Immune reconstitution in patients underwent HI-BMT was later than that in other two groups. In 2 patients TCRBV was detected in 2m and 3m after allo-HSCT and found that there was a tendency of increasing usage of BV subfamilies and increasing expression of CDR3 polymorphism. Twenty three TCRBV CDR3 molecules associated with GVHD and CMV infection were compared each other by bioinformatics and found that different cases of the same BV subfamilies may share similarity in amino acid motif, while in different BV subfamilies none appeared to share the same amino acid motif.
In 1.5 years after allo-HSCT, the usage of TCRBV subfamilies still restricted. Immune reconstitution in patients underwent HI-BMT was later than that in other two groups. TCRBV CDR3 molecules associated with GVHD and CMV infection showed that different cases of the same BV subfamilies may share similarity in amino acid motif, while in different BV subfamilies none of clones appeared to share the same amino acid motif.
研究异基因造血干细胞移植（allo-HSCT）后白血病受者T淋巴细胞克隆的T细胞受体β链可变区（TCRBV）的CDR3库的分子特征。
采用逆转录聚合酶链反应（RT-PCR）从24例接受三种allo-HSCT的白血病患者外周血（PB）淋巴细胞及5例正常供者作为对照中扩增TCRBV的24个亚家族基因。PCR产物进一步通过基因扫描分析以评估BV亚家族的克隆性及CDR3的特征，并计算BV亚家族的使用率。通过变性聚丙烯酰胺凝胶电泳获得与移植物抗宿主病（GVHD）和巨细胞病毒（CMV）感染相关的单克隆条带并进行测序。报道了TCRBV CDR3序列与其他与GVHD或CMV感染相关的CDR3序列的比较。
移植后约2至19个月，9例接受单倍体相合骨髓移植（HI-BMT）的患者中有6至14个BV亚家族表达，多克隆表达率达33%。5例接受非血缘外周血干细胞移植（MU-PBSCT）的患者中有10至15个BV亚家族表达，其中45%为多克隆。10例接受同胞相合骨髓移植（MS-BMT）的患者中有10至16个BV亚家族表达，其中超过48%为多克隆。24个BV亚家族中存在单克隆和寡克隆，但未发现共同的单克隆BV亚家族。接受HI-BMT的患者免疫重建晚于其他两组。2例患者在allo-HSCT后2个月和3个月检测到TCRBV，发现有BV亚家族使用率增加及CDR3多态性表达增加的趋势。通过生物信息学比较了23个与GVHD和CMV感染相关的TCRBV CDR3分子，发现同一BV亚家族的不同病例在氨基酸基序上可能具有相似性，而不同BV亚家族中没有克隆似乎共享相同的氨基酸基序。
allo-HSCT后1.5年内，TCRBV亚家族的使用仍受限。接受HI-BMT的患者免疫重建晚于其他两组。与GVHD和CMV感染相关的TCRBV CDR3分子显示，同一BV亚家族的不同病例在氨基酸基序上可能具有相似性，而不同BV亚家族中没有克隆似乎共享相同的氨基酸基序。
