Rukavina D, Balen-Marunic S, Rubesa G, Orlic P, Vujaklija K, Podack E R
Department of Physiology and Immunology, Medical Faculty, University of Rijeka, Croatia.
Transplantation. 1996 Jan 27;61(2):285-91. doi: 10.1097/00007890-199601270-00021.
Perforin (P) is a cytolytic molecule expressed in the granules of cytolytic T cells and natural killer cells. Although cytotoxic cells have been implicated in graft rejection, no prospective clinical study has been published that examines the dynamics of perforin expressing cells in peripheral blood lymphocytes of transplanted patients. The cytofluorimetric assay developed in our laboratory previously for the simultaneous detection of intracellular perforin together with cell surface molecules was used for posttransplantation monitoring of patients, for the assessment of the efficiency of immunosuppressive treatment, and for the prediction of acute kidney transplant rejection and the stability of tolerance to long lived kidney transplants. Immunosuppression for the purpose of allotransplantation causes a decline in the number of perforin-expressing cells in peripheral blood. In contrast, in patients with clinical signs of acute rejection, the total number of perforin-expressing lymphocytes was increased in comparison with nonrejecting patients. Analyzing perforin-expressing subsets, rejection crises were accompanied by a relative decrease of perforin expression in the CD4+ subpopulation while increasing in the CD8+ subset. In the CD56+ and CD16+ NK subpopulations changes in perforin expression were mixed. In nonrejecting patients the ratio of perforin expression in CD4+ cells was high compared with CD8+ cells. Intensive therapy of acute rejection episodes with high doses of corticosteroids (methylprednisolonet [Solumedrol] bolus) strongly and significantly decreased the percentage of both, the subpopulations of perforin-positive T cells and the subpopulation of CD56+P+ NK cells. The lowest level of perforin expression, including low frequencies of perforin among CD8+ and CD4+ cells, was found in the group of patients tolerating transplanted kidneys for several years. These changes in perforin protein expression in peripheral blood can be used to discriminate between immunosuppressed patients who are immunologically quiescent and those who undergo transplant rejection. Our results confirm the hypothesis that cytotoxicity mediated by perforin may be an important effector mechanism in the rejection of allografted kidneys.
穿孔素（P）是一种在细胞毒性T细胞和自然杀伤细胞的颗粒中表达的溶细胞分子。尽管细胞毒性细胞与移植物排斥反应有关，但尚未发表前瞻性临床研究来检测移植患者外周血淋巴细胞中穿孔素表达细胞的动态变化。我们实验室之前开发的用于同时检测细胞内穿孔素和细胞表面分子的细胞荧光分析方法，被用于移植后患者的监测、免疫抑制治疗效果的评估、急性肾移植排斥反应的预测以及对长期存活肾移植耐受性的稳定性评估。同种异体移植目的的免疫抑制会导致外周血中穿孔素表达细胞数量减少。相反，有急性排斥临床症状的患者中，与未发生排斥的患者相比，穿孔素表达淋巴细胞的总数增加。分析穿孔素表达亚群发现，排斥危象伴随着CD4 +亚群中穿孔素表达相对减少，而CD8 +亚群中穿孔素表达增加。在CD56 +和CD16 +自然杀伤细胞亚群中，穿孔素表达的变化是混合的。在未发生排斥的患者中，CD4 +细胞中穿孔素表达与CD8 +细胞相比比例较高。用高剂量皮质类固醇（甲泼尼龙[甲强龙]推注）对急性排斥发作进行强化治疗，可显著降低穿孔素阳性T细胞亚群和CD56 + P +自然杀伤细胞亚群的百分比。在耐受移植肾数年的患者组中，发现穿孔素表达水平最低，包括CD8 +和CD4 +细胞中穿孔素频率较低。外周血中穿孔素蛋白表达的这些变化可用于区分免疫静止的免疫抑制患者和发生移植排斥的患者。我们的结果证实了穿孔素介导的细胞毒性可能是同种异体移植肾排斥反应中一种重要效应机制的假设。
