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目的探讨急性排斥反应发生的机制,寻找适合临床早期诊断急性排斥反应(AR)的敏感指标。方法采用免疫组织化学法检测移植肾组织中穿孔素和颗粒酶B的表达;利用流式细胞仪检测法和免疫细胞化学ABC法,检测外周血淋巴细胞中穿孔素和颗粒酶B的表达。结果AR的移植肾病理标本均100%表达穿孔素和颗粒酶B,二者的表达情况类似,主要定位于肾小管上皮细胞和间质淋巴细胞的胞浆和胞膜,与病理分级呈正相关,颗粒酶B的相关程度更好。发生AR的患者外周血淋巴细胞中穿孔素和颗粒酶B的表达较肾功能稳定的患者有明显的升高,而且在出现明显的AR临床症状前3d就已经开始升高,经过有效的抗排斥治疗后,表达明显减弱或转阴。结论监测外周血淋巴细胞中穿孔素和颗粒酶B的表达水平,可做为早期发现AR的一种检测方法。穿孔素和颗粒酶B可以反映急性排斥的病情,对移植肾的预后有一定的指导价值。
Objective To investigate the mechanism of acute rejection and find the sensitive index suitable for early diagnosis of acute rejection (AR). Methods The expression of perforin and granzyme B in renal allograft tissues was detected by immunohistochemistry. The expression of perforin and granzyme B in peripheral blood lymphocytes was detected by flow cytometry and immunocytochemical ABC method. Results The pathology of renal allografts in AR showed 100% expression of perforin and granzyme B. The expression of perforin and granzyme B were similar and mainly located in the cytoplasm and membrane of renal tubular epithelial cells and interstitial lymphocytes, which was positively correlated with pathological grade, Granzyme B is more relevant. Peripheral blood lymphocytes in patients with AR expression of perforin and granzyme B than in patients with stable renal function was significantly increased, but also in the obvious AR clinical symptoms before 3d began to rise, after effective anti-rejection After treatment, the expression was significantly weakened or turned negative. Conclusion Monitoring perforin and granzyme B expression in peripheral blood lymphocytes can be used as a detection method for early detection of AR. Perforin and granzyme B can reflect the acute rejection of the disease, the prognosis of renal transplantation has some guiding value.