论文部分内容阅读
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 effect of sepsis and other diseases on thymus output and to establish a method for detecting T cell receptor deletion loop (TREC). Methods The TREC plasmid standard was constructed and the real-time quantitative PCR method was used to draw the standard curve. Blood samples from healthy people of different ages were selected and their peripheral blood mononuclear cells (PBMCs) were extracted. T cells of different subpopulations were sorted out by flow cytometry to detect the content of TREC (copy / Tcell). Results TREC was detected in different T cell subsets of 10 PBMCs from healthy donors. The mean levels of TREC in CD4 ~ + CD45RA ~ + and CD8 ~ + CD45RA ~ + naive T cell subsets were significantly higher than those in the corresponding CD45RO ~ + memory T cells (P <0.05) With age, it shows a gradual downward trend. Conclusion The method of real-time quantitative PCR for detecting TREC was established to evaluate the output of thymus T cells.