INVESTIGATION ON THE REDUCTION OF IMMUNOSUPRESSANTS IN LONG TERM SURVIVED POST RENAL TRANSPLANT PATIENTS.

白细胞介素2(IL-2)由辅助性T细胞产生，在移植后的免疫调节中起重要作用，IL-2基因的调控元件有两个AP-1和NF-AT结合位点。最近有报道称，与正常T细胞相比，无能T细胞的转录因子AP-1与核因子-AT和核因子-AT的结合活性降低和下调。4例正常人、4例存活5年以上的肾移植受者和4例存活5年以下的肾移植受者外周血经Ficoll梯度离心法提取淋巴细胞，在37ﾟC条件下分别培养4h，分别在不同浓度的PMA和PHA刺激下培养4h，以评价免疫抑制剂的免疫调节作用更多的艾尔。将IL-2基因启动子区域的AP-1和NF-AT共同序列末端标记的DNA片段与核抽提物反应。4名正常志愿者AP-1和NF-AT的DNA结合活性处于中等水平，而4名长期存活的肾移植受者的AP-1和NF-AT的DNA结合活性较低。此外，在这些淋巴细胞中检测到PMA和PHA的刺激作用；而所有四名长期存活的受者都没有表现出AP-1和NF-AT的结合活性增强，而四名正常志愿者中的两名表现出增强作用。在本文中，我们证明了免疫抑制状态下AP-1和NF-AT的DNA结合活性降低，甚至在PMA和PHA刺激后也没有增强。对AP-1和NF-AT结合活性的研究将为我们提供免疫抑制效应的估计，从而建立更常用的免疫抑制方案。较少

Interleukin-2 (IL-2) produced by helper T cells has been showm to play an important role in the immunomodulation after transplantation and regulatory elements of IL-2 gene have two AP-1 and NF-AT binding sites. Recently it has been reported that binding activities of transcription factor AP-1 and NF-AT and NF-AT of anergic T cell are decreased and down-regulated as compared with normal T cells. In this study, we have assessed DNA-binding activities of AP-1 and NF-AT by electrophoretic mobility shift assay in posttransplant recipients in order to evaluate the immunomodulatory effect of the immunosuppressants.Lymphocytes were prepared from perlpheral blood of four normal volunteers, four renal transplant recipients surviving for more than five years and four that surviving less than 5 years by gradient centrifugation over Ficoll and cultured for four hours at 37ﾟC with or without stimulation of PMA and PHA.Nuclear extracts were prepared from lympocytes by the modified method of Dignam et … More al. Endlabeled DNA fragments of AP-1 and NF-AT binding consensus sequences of IL-2 gene promoter regions were reacted with nuclear extracts. Electrophoretic mobility shift assays were performed and BAS-2000 Bioimage analyzer was used for the detection of DNA binding complex of AP-1 and NF-AT.Four normal volunteers showed DNA-binding activities of AP-1 and NF-AT in moderate level, but four long term surviving renal transplant recipients showed much lower level of the activities. Furthermore, a stimulatory effect of PMA and PHA was examined in these lymphocytes ; whereas all of four long term surviving recipients showed no enhancement of binding activities of AP-1 and NF-AT,two of four normal volunteers showed enhancements.In this paper, we have shown that the DNA-binding activities of AP-1 and NF-AT in immumosuppressive state, are decreased and not enhanced even after the stimulation of PMA and PHA.Investigations of the binding activivies of AP-1 and NF-AT will provide us the estimation of immunosuppressive effect, leading to establishment of more usuful immunosuppressive regimen. Less