MLC上清液中细胞因子的检测与肾移植急性排斥反应的研究

An experimental study on the relationship between cytokines in MLC supernatants and acute rejection after renal transplantation

目的 探讨混合淋巴细胞培养 (MLC)中细胞因子水平与急性排斥反应的关系。方法随机选取 2 4例接受同种异体肾移植术的患者为检测对象。在其接受手术当日 ,分离其外周血单个核细胞 (PBMC)与供者单个核细胞进行混合白细胞培养 (反应组 ) ,同时以单纯自身PBMC进行培养 (对照组 )。在培养第 1、3、5d测MLC上清中IL 2、IL 6和TNF α水平 ,并观察其分泌动态变化。结果 发生急性排斥反应的患者 ,在 3次时间点上 ,细胞因子水平均高于肾功能良好组 ,差异达到显著水平。反应组较对照组细胞因子水平有增加 ,急排的病人增加量高于肾功能良好的受者 ,差异达到显著水平。排斥组与肾功能良好组 3种细胞因子的分泌规律基本一致 ,但峰值都有增高。结论 检测MLC中细胞因子水平可以作为合理选择供受者、预测急性排斥的方法。并启发利用抗细胞因子抗体逆转急性排斥反应。

Objective Investigate the relationship between cytokine level in MLC supernatant and acute rejection after renal transplantation. Methods 17 male patients and 7 female patients who received renal allografts were chosen randomly. PBMCs were separated routinely from donors and recipients. For one way MLC, stimulator PBMCs were treated with mitomycin. Cell concentration was 1.5×10 6/ml. Responder PBMCs (recipients' PBMC ) were co cultured with either mitomicin c treated autologous PBMCs (control cultures) or mitomicin c treated allogeneic (stimulated cultures) in replicates of four. For the quantitative determination of IL 2, IL 6 and TNF α, we used ELISA. The results were measured on day 1, 3 and 5. Results In 10 patients who suffered acute rejection(RG group), the levels of all cytokines in control and stimulated cultures were higher than those of 14 patients whose renal function was stable (NG group). The difference was significant. Either in RG or NG on day 1, 3 and 5 all cytokines' levels in stimulated cultures were higher than control cultures. The kinetics of the production of IL 2, and TNF α in RG were similar to that of NG. In RG, the peak value of IL 6 appeared one day earlier than in NG. Conclusions 1. IL 2, IL 6 and TNF α, as effectors, play critical roles in cell mediated transplantation immunity. The enhancement of their levels in MLC supernatants may predict the occurrence of acute rejection. 2. MLC has the predictive value for transplantation success. It may also be useful for the selection of most compatible related and unrelated donors.