Molecular markers in subclinical acute rejection of renal transplants
Molecular markers in subclinical acute rejection of renal transplants
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DOI：
10.1111/j.1399-0012.2004.00161.x
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发表时间：
2004-06-01
影响因子：
2.1
通讯作者：
Manfro, RC
中科院分区：
文献类型：
--
作者：
Dias, ECA;Veronese, FJV;Manfro, RC
在这项研究中，我们评估了急性排斥反应分子标志物在有和没有亚临床急性排斥反应（SAR）的患者的方案活检中的表达。在肾移植后2个月（n = 21）和12个月（n = 14）对异体移植物功能稳定的患者进行方案活检。活检组织RNA分离后，对甘油醛3-磷酸脱氢酶（GAPDH）、穿孔素、颗粒酶B和Fas配体基因进行逆转录和聚合酶链反应（RT-PCR）。采用Banff 97分级进行组织学诊断。SAR患者2个月时肌酐浓度显著升高（1.46 +/- 0.27 x 1.18 +/- 0.24; p < 0.02）。15例活检标本中发现穿孔素转录物，其中10例有SAR的组织学征象（p = 0.06）。颗粒酶B在10份标本中表达，其中9份呈SAR （p < 0.01）。7个标本中表达Fas配体，其中6个归为SAR （p < 0.01）。穿孔素表达对SAR诊断的敏感性最高（81%），颗粒酶B和Fas配体的特异性为90%。在12个月时，有和没有既往SAR患者的肌酐浓度无显著差异（1.63 +/- 0.57 x 1.28 +/- 0.31; p = 0.10）。分子分析结果显示，有SAR与无SAR患者的穿孔素和颗粒酶B的表达差异无统计学意义。5个样本中观察到Fas配体表达，其中4个有SAR的组织学征象（p = 0.03）。在12个月时，对SAR的诊断，穿孔素表达的敏感性最高（83%），Fas配体的特异性最高（88%）。我们得出结论，SAR肾脏中，参与对同种异体移植物的细胞溶解攻击的编码蛋白的基因表达增加。这些发现支持了SAR是一种对同种异体肾脏移植物潜在有害的主动免疫过程的理解。
In this study, we evaluated the expression of molecular markers of acute rejection in protocol biopsies of patients with and without subclinical acute rejection (SAR). Protocol biopsies were performed at 2 months (n = 21) and 12 months (n = 14) after kidney transplantation in patients with stable allograft function. After biopsy tissue RNA isolation, reverse transcription and polymerase chain reaction (RT-PCR) for the glyceraldehyde 3-phospate dehydrogenase (GAPDH), perforin, granzyme B and Fas ligand genes were performed. The Banff 97 classification was used for histological diagnosis. Creatinine concentrations at 2 months were significantly higher in patients with SAR (1.46 +/- 0.27 x 1.18 +/- 0.24; p < 0.02). Perforin transcripts were found in 15 biopsy specimens, 10 of which had histological signs of SAR (p = 0.06). Granzyme B expression was found in 10 specimens, nine of which had SAR (p < 0.01). Fas ligand was expressed in seven specimens, and six of them were classified as SAR (p < 0.01). Perforin expression had the highest sensitivity (81%) for the diagnosis of SAR. Granzyme B and Fas ligand had specificity of 90%. At 12 months, there was no significant difference in creatinine concentrations for patients with and without previous SAR (1.63 +/- 0.57 x 1.28 +/- 0.31; p = 0.10). Molecular analysis revealed that there was no statistically significant difference in the expression of perforin and granzyme B in patients with and without SAR. Fas ligand expression was observed in five samples, four of which had histological signs of SAR (p = 0.03). At 12 months, perforin expression had the highest sensitivity (83%), and Fas ligand, the highest specificity (88%) for the diagnosis of SAR. We concluded that the expression of genes that encode proteins involved in the cytolytic attack against the allograft is increased in kidneys with SAR. These findings support the understanding that SAR is an active immune process potentially deleterious to renal allografts.