CML患者胸腺近期输出功能和TCR Vβ谱系利用特点.docVIP

中文摘要 目的：了解CML患者胸腺近期输出功能（TRECs的含量）以及TCR Vβ亚家族T 细胞的基因表达和克隆性增殖情况，从而为临床特异性免疫治疗研究提供资料。 方法：收集38例CML患者的42份外周血标本，免疫荧光检测的PBMCs中CD3+细胞的阳性率，计算CD3+细胞中TRECs含量RT-PCR和基因扫描方法分析CML患者PBMCs、CD4+和CD8+细胞中TCR Vβ 24个亚家族T细胞的基因表达和克隆性。14例正常人外周血作对照。 结果：CML-CP患者PBMCs、CD3+、CD4+和CD8+细胞中TRECs含量均明显低于正常人，而缓解期和allo-BMT 6个月后的患者CD3+细胞中TRECs含量比发病期有所升高，比正常人稍低，但差别均无统计学意义。不同性别的正常人和CML-CP患者外周血TRECs含量也有差别，正常人CD8+细胞和CML-CP患者CD4+细胞中TRECs含量，女性均明显高于男性，而在其它各组，尽管女性TRECs含量也比男性高，但差异无统计学意义。 全部CML患者均存在不同程度的Vβ亚家族缺失5.52个（PBMCs）、11.58个CD4+）和12.37 个CD8+）Vβ亚家族，以Vβ13的表达率最高Vβ亚家族的表达明显减少，达完全缓解时，Vβ亚家族的表达逐渐恢复。基因扫描分析发现：PBMCs、CD4+和CD8+细胞19/27例（14/19例）89.5％（17/19例）部分Vβ亚家族克隆T细胞Vβ亚家族以Vβ21克隆性增殖最常Vβ亚家族Vβ11的克隆性增殖最常见。进一步分析还发现TCR Vβ亚家族TRECs 实时定量PCR TCR Vβ亚家族 基因扫描 Abstract Objective: To investigate the thymic recent output function (content of TRECs) and the gene expression of TCR Vβ subfamily T cells and clonality in patients with chronic myelogenous leukemia(CML). This study is expected to provide some basic data for research on clinical specific immune therapeusis. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from 42 heparinized peripheral blood samples obtained from 38 patients. Some of PBMCs were used to separate CD4+ and CD8+ cells by using the MACS magnetic bead separation system. Quantitative detection of T-cell receptor excision DNA circles (TRECs) in DNA of PBMCs, CD4+and CD8+cells were preformed by real-time PCR analysis. And the TREC-number was related to the number of T-cells by determination of the number of CD3-positive cells. The gene expression and cloanlity analysis of TCR Vβ repertoire were detected by RT-PCR and genescan technique in PBMCs, CD4+ and CD8+ cells. 14 normal individuals served as controls. Results: There were dramatic reduction of TREC levels in PBMCs, CD3+, CD4+ and CD8+ cells in patients with CML-CP compared with normal individuals. Whereas the TREC levels in CD3+ cells from patients with CML who achieved complete remission or stayed at 6 months after allo-BMT were higher