Quantitative detection of T-cell activation markers by real-time PCR in renal transplant rejection and correlation with histopathologic evaluation
Quantitative detection of T-cell activation markers by real-time PCR in renal transplant rejection and correlation with histopathologic evaluation
复制标题
DOI：
10.1097/00007890-200209150-00019
复制
发表时间：
2002-09-15
期刊：
影响因子：
6.2
通讯作者：
Gaber, L
中科院分区：
文献类型：
--
作者：
Sabek, M;Dorak, MT;Gaber, L
背景最近的观察表明，特异性细胞毒性T细胞标志物在排斥反应中上调，这使得对诊断实体器官移植排斥反应的非侵入性方法的探索重新焕发了活力。我们开发了一种一步实时聚合酶链反应（PCR）方法，可以在相对较短的时间内可靠地检测几种T细胞基因的表达。该测定具有高度灵敏性和可重复性，具有宽的动态范围，允许在低至3 pg的总RNA中准确定量靶mRNA。评价了该方法在检测肾移植排斥反应中的实用性。从27例因移植后肾功能不全而接受肾移植活检的患者中收集外周血单个核细胞。T细胞活化标志物颗粒酶B、穿孔素和HLA-κ B的表达被定量并与肾活检的组织病理学变化相关。排斥反应组（n=8）外周血淋巴细胞颗粒酶B表达明显增高（P
Background. The quest for noninvasive methods to diagnose rejection in solid-organ transplants has been rejuvenated by recent observations that specific cytotoxic T-cell markers are up-regulated during rejection.Methods. We developed a one-step real-time polymerase chain reaction (PCR) method allowing reliable detection of the expression of several T-cell genes within a relatively short period of time. The assay is highly sensitive and reproducible with a wide dynamic range allowing accurate quantification of target mRNA in as little as 3 pg total RNA. The utility of this assay in detecting renal allograft rejection was evaluated. Peripheral blood mononuclear cells were collected from 27 patients undergoing kidney allograft biopsies for renal dysfunction after transplantation. Expression of the T-cell activation markers, granzyme B, perforin, and HLA-DRA, was quantified and correlated to the histopathologic changes in the renal biopsies.Results. In cases with allograft rejection (n=8), peripheral lymphocyte expression was increased for granzyme B (P