Evaluation of two color flow cytometric analysis of peripheral immunocompetent cell phenotypes as an immunomonitoring for kidney allografted patients

对肾移植患者的管理需要可靠的免疫学监测方法。用单抗流式细胞术分析外周血免疫活性细胞表型是一种理想的监测方法。在本研究中，我们评价了单色和双色流式细胞术外周血单核细胞表型分析在移植患者管理中的应用。对41例肾移植患者、34例慢性肾功能衰竭透析患者和41例健康成人外周血有核细胞进行了检测。用PE标记的Leu系列单抗和流式细胞仪(FACSIV)分析细胞表型。在双色实验中，我们对激活的PAN T(Leu4DR)、激活的PAN T(Leu4I12)、激活的T抑制/杀伤(Leu2aDR)、T抑制(Leu2a15)、T杀伤(Leu2a1-)、T抑制诱导因子(Leu3a8-)、辅助性T细胞(Leu3a8-)、激活的单核细胞(LeuDRM3)进行了定量移植后12个月移植患者高于正常对照组。无急性排斥反应的移植患者T细胞抑制率高于有急性排斥反应的患者，尤其是术后3~12个月。移植后12个月T抑制诱导物低于正常对照组和慢性肾功能衰竭患者，活化的单核细胞数高于正常对照组和慢性肾功能衰竭患者。不同免疫抑制剂对亚群比例无明显影响。时间序列分析未发现细胞亚群的改变与实验室数据或临床病程之间有任何共同的相关性。目前，我们的结论是，外周免疫活性细胞亚群的定量在移植患者的管理中并不是很有用。我们建立了用单抗、荧光乳胶珠和流式细胞术检测巨噬细胞吞噬活性的新方法。现在，我们正在研究这一新的检测方法，为移植患者的管理提供有用的监测。较少

Reliable immunological monitoring method useful for the management of kidney transplanted patients has been required. Flowcytometric analysis of peripheral immunocompetent cell phenotypes using monoclonal antibody had been expected to be an ideal monitoring method.In this study, we evaluated the single color and two color flowcytometric analysis of peripheral mononuclear cell phenotypes in the management of tranplant patients. Peripheral nucleated cells of 41 kidney transplanted patients, 34 dialyzed chronic renal failure patients, 41 healty adults were examined. Cell phenotypes were analyzed using Leu series monoclonal antibodies conjugated FITC of PE and flowcytomter(FACS IV). In two color assay, we quantitated activated pan T (Leu4+DR+), activated pan T (Leu4+I12+), activated T suppressor/killer(Leu2a+DR+), T suppressor(Leu2a+15+), T killer(Leu2a+1-), T suppressor inducer(Leu3a+8+), T helper(Leu3a+8-), activated monocyte(LeuDR+M3+).Activated T suppressor/killer ratio in total cells … More of transplanted patients were higher than normal control 12 month after transplantation. T suppressor ratio of transplanted patients without acute rejection were higher than those with it, especially in the period between 3 to 12 months after surgery.T suppressor inducer 12 months after transplantation was lower and activated monocytes of transplanted patients was higher than normal control and chronic renal failure patients. Kinds of immunosuppressant did not paticular affect on the subsets ratio. Time series analysis could not find any common correlation between the alteration of cell subsets and laboratory data or clinical course. At present, we concluded that the quantitation of peripheral immunocompetent cell subsets were not so useful in management of transplanted patients. We developed new assay of phagocytic activity of macrophage using monoclonal antibody, fluorescence latex beads and flowcytometry. Now we are studying this new assay for useful monitoring for management of transplanted patients. Less