体外刺激肾移植受者外周血淋巴细胞进行免疫监测的临床研究

目的探讨应用体外刺激外周血淋巴细胞的方法进行免疫监测来预测肾移植术后急性排斥反应的价值。方法选取40例肾移植受者,将其分为急性排斥反应组(20例)和肾功能正常组(20例);选取10名健康供者作为正常对照组。抽取所有研究对象的外周静脉血,分离出外周血淋巴细胞,并经佛波酯(phorbol myfismte acetate,PMA)和离子霉素(ionomycin,Ion)体外刺激8h。应用流式细胞术检测细胞培养上清液白介素(IL)-2、IL-6的表达强度及CD3+CD69+、CD3+CD71+淋巴细胞比例;应用受试者工作特征(ROC)曲线评价IL-2联合IL-6、CD3+CD69+联合CD3+CD71+预测肾移植术后急性排斥反应的敏感度和特异度。结果急性排斥反应组的IL-2、IL-6的荧光强度和CD3+CD69+、CD3+CD71+淋巴细胞比例均显著高于肾功能正常组(P≤0.01);IL-2联合IL-6预测肾移植受者急性排斥反应的敏感度为0.90,特异度为0.95。CD3+CD69+联合CD3+CD71+预测急性排斥反应的敏感度为0.95,特异度为0.90。结论监测肾移植受者外周血淋巴细胞体外刺激上清液中的IL-2、IL-6及CD3+CD69+、CD3+CD71+的表达强度,可以预测急性排斥反应。此法简便,敏感度和特异度高。

Clinical research of peripheral blood lymphocytes stimulated in vitro for immune monitoring in renal transplant recipients

Objective To explore the value of using peripheral blood lymphocytes stimulated in vitro for immune monitoring to predict acute rejection in kidney transplant recipients.Methods Forty kidney transplant recipients were selected as objects and divided into acute rejection group(n=20)and normal renal function group(n=20).And 10 healthy donors were selected as control group.Peripheral blood lymphocytes of all objects were cultured and stimulated in vitro by phorbol myfismte acetate(PMA)/ionomycin(Ion) for 8 hours.The expression of interleukin(IL)-2,IL-6 in cell culture supernatant and CD3+CD69+,CD3+CD71+ lymphocytes were detected by flow cytometry.The sensitivity and specificity of the method of combining IL-2,IL-6 or CD3+CD69+,CD3+CD71+ on predicting acute rejection after kidney transplantation were analyzed by the curve of receiver operating characteristic(ROC).Results The fluorescence intensity of IL-2,IL-6 and ratio of CD3+CD69+,CD3+CD71+ lymphocytes in normal renal function group was significantly lower than that in acute rejection group(P≤0.01).The sensitivity and specificity of IL-2 combined IL-6 for predicting acute rejection of kidney transplant recipients was respectively 0.90 and 0.95.And the sensitivity and specificity of CD3+CD69+ combined CD3+CD71+ lymphocytes for predicting acute rejection were respectively 0.95 and 0.90.Conclusions Monitoring the expression of cell culture supernatant IL-2 combined IL-6 and ratio of CD3+CD69+ combined CD3+CD71+ lymphocytes after stimulating the peripheral blood lymphocytes in vitro of kidney transplant recipients can predict acute rejection,which is a simple method with high sensitivity and specificity.