Strehlau J, Pavlakis M, Lipman M, Shapiro M, Vasconcellos L, Harmon W, Strom T B
Department of Medicine, Harvard Medical School, Boston, MA 02215, USA.
Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):695-700. doi: 10.1073/pnas.94.2.695.
Procedures to diagnose renal allograft rejection depend upon detection of graft dysfunction and the presence of a mononuclear leukocytic infiltrate; however, the presence of a modest cellular infiltrate is often not conclusive and can be detected in non-rejecting grafts. We have pursued a molecular approach utilizing reverse transcription (RT)-PCR to test the diagnostic accuracy of multiple immune activation gene analysis as means to diagnose renal allograft rejection. The magnitude of intragraft gene expression of 15 immune activation genes was quantified by competitive RT-PCR in 60 renal allograft core biopsies obtained for surveillance or to diagnose the etiology of graft dysfunction. Results were compared with a clinicopathological analysis based upon the histological diagnosis (Banff criteria) and the response to antirejection treatment. During acute renal allograft rejection intragraft expression of the interleukin (IL)-7 (P < 0.001), IL-10 (P < 0.0001), IL-15 (P < 0.0001), Fas ligand (P < 0.0001), perforin (P < 0.0001), and granzyme B (P < 0.0015), but not IL-2, interferon gamma, or IL-4, genes is significantly heightened. Amplified RANTES and IL-8 gene transcripts are sensitive but nonspecific markers of rejection. A simultaneous RT-PCR evaluation of perforin, granzyme B, and Fas ligand identifies acute rejection, including cases with mild infiltration, with extraordinary sensitivity (100%) and specificity (100%). Effective antirejection therapy results in a rapid down-regulation of gene expression. The combined analysis of Fas ligand, perforin, and granzyme B gene expression by quantitative RT-PCR provides a reliable tool for diagnosis and follow-up of acute renal allograft rejection. Its accuracy and a potential rapid application within few hours suggest its use in the clinical management of renal transplant patients.
诊断肾移植排斥反应的程序依赖于对移植肾功能障碍的检测以及单核白细胞浸润的存在；然而，适度的细胞浸润的存在往往并不具有决定性，并且在未发生排斥反应的移植肾中也可检测到。我们采用了一种分子方法，利用逆转录（RT）-PCR来测试多种免疫激活基因分析作为诊断肾移植排斥反应手段的诊断准确性。通过竞争性RT-PCR对60例用于监测或诊断移植肾功能障碍病因的肾移植核心活检组织中15种免疫激活基因的移植内基因表达量进行定量。将结果与基于组织学诊断（班夫标准）和抗排斥治疗反应的临床病理分析进行比较。在急性肾移植排斥反应期间，白细胞介素（IL）-7（P < 0.001）、IL-10（P < 0.0001）、IL-15（P < 0.0001）、Fas配体（P < 0.0001）、穿孔素（P < 0.0001）和颗粒酶B（P < 0.0015）基因的移植内表达显著升高，但IL-2、干扰素γ或IL-4基因未升高。趋化因子调节激活正常T细胞表达和分泌（RANTES）和IL-8基因转录本的扩增是排斥反应的敏感但非特异性标志物。同时进行穿孔素、颗粒酶B和Fas配体的RT-PCR评估可识别急性排斥反应，包括轻度浸润的病例，具有极高的敏感性（100%）和特异性（100%）。有效的抗排斥治疗导致基因表达迅速下调。通过定量RT-PCR对Fas配体、穿孔素和颗粒酶B基因表达进行联合分析，为急性肾移植排斥反应的诊断和随访提供了一种可靠的工具。其准确性以及在数小时内潜在的快速应用表明其可用于肾移植患者的临床管理。
