南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (03): 294-296.

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三色和双色荧光标记流式细胞术对肾移植免疫反应新指标TLR4的监测

于立新； 余玉明； 邓文锋； 刘艳君；   

  1. 南方医科大学南方医院器官移植科； 南方医科大学免疫学教研室 广东广州510515； 广东广州510515；
  • 出版日期:2006-03-20 发布日期:2006-03-20
  • 基金资助:
    广东省自然科学基金(23003)~~

Three-and two-color flow cytometry for immunologic surveillance of toll-like receptor 4 after renal transplantation

YU Li-xin1,YU Yu-ming1,DENG Wen-feng1,LIU Yan-jun2 1Department of Organ Transplantation,Nanfang Hospital;2Department of Immunology,Southern Medical University,Guangzhou 510515,China   

  1. 南方医科大学南方医院器官移植科； 南方医科大学免疫学教研室 广东广州510515； 广东广州510515；
  • Online:2006-03-20 Published:2006-03-20

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摘要： 目的比较三色和双色荧光标记流式细胞术(FCM)在肾移植术后免疫指标TLR4监测中的优缺点。方法选取10例肾移植术后病例,分别采用三色和双色FCM检测外周血单核细胞中CD14、TLR4和CD80表达情况,计算CD14阳性单核细胞中TLR4和CD80的表达百分率并进行统计比较。结果两种方法检测CD14阳性单核细胞中TLR4和CD80阳性表达百分率均无显著性差异(P=0.198,P=0.872),两种方法对相同血标本的检测结果均呈正相关关系(积矩相关系数r=1,P=0.000;积矩相关系数r=0.999,P=0.000)。另外,应用三色FCM还可同时获得单核细胞中TLR4和CD80双阳性的表达情况。结论三色和双色FCM检测TLR4的结果无显著性差异,但三色FCM可同时显示TLR4与其下游分子CD80双阳性表达情况,对揭示TLR4参与移植免疫反应机制具有重要价值,而且血标本用量少,节约单克隆抗体。 更多还原

Abstract: Objective To compare the characteristics of three-and two-color flow cytometry for immunologic surveillance of toll-like receptor 4(TLR-4) after renal transplantation.Method The expressions of CD14,TLR4 and CD80 in peripheral blood monocytes were measured in 10 patients after renal transplantation by three-and two-color fluorescent staining flow cytometry,respectively.The percentages of TLR4 and CD80 in CD14-positive monocytes were calculated,and the characteristics of two methods of flow cytometry were compared.Results The two detection methods did not significantly differ in their detection of TLR4(P=0.198) and CD80(P=0.872) expressions in CD14-positive monocytes,and the measurements of the two methods for the same sample showed positive correlation(r=1,P=0.000 for TLR4 and r=0.999,P=0.000 for CD80).Three-color flow cytometry was also capable of detecting monocytes positive for both TLR4 and CD80.Compared with two-color flow cytometry,three-color flow cytometry provided more information and required less blood sample and monoclonal antibody.Conclusions Three-and two-color flow cytometry have almost the same accuracy in immunologic surveillance of TLR4 after renal transplantation.The former method also provides information of the simultaneous expression of TLR4 and its down stream molecule CD80,which could be critical for imploring the mechanisms by which TLR4 takes part in alloimmunity through modulating CD80 expression,and requires less blood sample and monoclonal antibody.

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