不同T细胞亚群T细胞受体删除环含量的评价

Evaluation of the content of T cell receptor excision circles in different T cell subpopulations

目的评价脓毒症等疾病对胸腺输出功能的影响,建立T细胞受体删除环(TREC)的检测方法。方法构建TREC质粒标准品,应用实时定量PCR法绘制标准曲线。选择不同年龄健康人的血液样本,提取其外周血单个核细胞(PBMCs),通过流式细胞术分选出不同亚群的T细胞,检测TREC含量(拷贝/Tcell)。结果 10例健康人血液PBMCs分选的不同T细胞亚群均检出TREC。CD4^+CD45RA^+及CD8^+CD45RA^+初始T细胞亚群中TREC平均含量均高于相应CD45RO^+记忆T细胞亚群的含量,差异具有统计学意义(P<0.05),且均随着年龄增长呈逐步下降趋势。结论建立实时定量PCR检测TREC的方法,用于评价胸腺T细胞的输出。

Objective To evaluate the influence of the resent thymic emigrant by sepsis disease, a method to detect T cell receptor excision circle （TREC） was found. Methods The TREC plasmid standard sample was established and the standard curve was drawn by real-time PCR. Peripheral blood mononuclear cells （PBMCs） were separated from healthy peripheral blood and T cell subpopulations were sorted by flow cytometer. Real-time PCR was applied to detect the content of TREC （copies/cell） in these T cell subpopulations. Results The detectable rate of TREC was 100% （10/10）. The content of TREC in CD4＋ CD45RA＋ T and CD8＋ CIM5RA＋ T cells were significantly higher than that in CD4＋ CD45RO＋ T ceils and CD8 ＋ CD45RO＋ T cells （P 〈 0.05 ）, respectively. The content of TREC in these T cell subpopulations gradually declined with the age increasing. Conclusions The method that could be applied to evaluate the recent thymic emigrant was established by detecting the content of TREC in T cell subpopulations with real-time PCR.