Desvaux Dominique, Schwarzinger Michaël, Pastural Myriam, Baron Christophe, Abtahi Mahdi, Berrehar François, Lim Annick, Lang Philippe, le Gouvello Sabine
Department of Pathology, Hôpital Henri Mondor, AP-HP 51, avenue du Marechal de Lattre-de-Tassigny, 94010 Créteil, France.
Transplantation. 2004 Sep 15;78(5):647-53. doi: 10.1097/01.tp.0000133530.26680.dc.
Because histopathologic criteria cannot always predict the pathogenesis and response to curative antirejection therapy, new hope derives from the molecular analysis of intragraft immunologic markers. We studied whether the cutoff of intragraft expression level of T-cell activation markers may define subgroups of acute rejection differing either in type of rejection or clinical outcome.
Forty-three human renal-allograft biopsies were quantified for mRNA expression of granzyme B, Fas ligand, interferon (IFN)gamma, interleukin (IL)-4, and IL-6 with a reverse-transcriptase real-time quantitative polymerase chain reaction (RT-PCR) method. Expression levels were correlated with the histopathologic rejection type according to the Banff 1997 classification criteria, and with the sensitivity to the antirejection immunosuppressive therapy, by means of receiver operating-characteristic (ROC) curves.
Granzyme B and Fas ligand mRNA expression up-regulation correlated with all allograft rejection types (P<0.01 for all). Moreover, granzyme B showed the highest sensitivity (90%) and specificity (78%) for the potential detection of histologic borderline changes that will require immunosuppressive therapy (area under the curve [AUC]=0.856, P<0.01). Curative antirejection-therapy resistance of overt, acute-rejection episode was significantly associated with higher Fas ligand gene expression (AUC=0.764, P<0.01, sensitivity [71%], specificity [99.5%]).
Real-time RT-PCR quantification of the over-expression of the granzyme B gene in kidney-graft biopsies has proved to be as reliable in detecting acute rejection as histologic assessment. Furthermore, we demonstrate that the simultaneous measurement of the mRNA up-regulation of Fas ligand might represent an efficient new tool for the prediction of pejorative outcome of acute rejection.
由于组织病理学标准无法总是预测发病机制以及对治愈性抗排斥治疗的反应，因此来自移植内免疫标志物分子分析的新希望应运而生。我们研究了移植内T细胞活化标志物表达水平的临界值是否可以定义在排斥类型或临床结果方面存在差异的急性排斥亚组。
采用逆转录实时定量聚合酶链反应（RT-PCR）方法对43例人肾移植活检组织中颗粒酶B、Fas配体、干扰素（IFN）γ、白细胞介素（IL）-4和IL-6的mRNA表达进行定量。根据1997年班夫分类标准，将表达水平与组织病理学排斥类型相关联，并通过受试者操作特征（ROC）曲线与对抗排斥免疫抑制治疗的敏感性相关联。
颗粒酶B和Fas配体mRNA表达上调与所有移植排斥类型相关（所有P<0.01）。此外，颗粒酶B对需要免疫抑制治疗的组织学临界变化的潜在检测显示出最高的敏感性（90%）和特异性（78%）（曲线下面积[AUC]=0.856，P<0.01）。明显急性排斥发作的治愈性抗排斥治疗抵抗与较高的Fas配体基因表达显著相关（AUC=0.764，P<0.01，敏感性[71%]，特异性[99.5%]）。
肾移植活检组织中颗粒酶B基因过表达的实时RT-PCR定量已被证明在检测急性排斥方面与组织学评估一样可靠。此外，我们证明同时测量Fas配体mRNA上调可能是预测急性排斥不良结局的一种有效新工具。
