Fernandez Luis A, Tsuchida Masahiro, Manthei Eric, Fechner John H, Oberley Terry D, Leverson Glen E, Knechtle Stuart J, Hamawy Majed M
Division of Transplantation, Department of Surgery, University of Wisconsin Medical School, 600 Highland Avenue, Madison, WI 53792-7375, USA.
Transpl Immunol. 2004 Sep-Oct;13(2):147-54. doi: 10.1016/j.trim.2004.05.007.
There is a need for a simple, sensitive, noninvasive technique for monitoring graft function. We report here on a simple assay called immune status assay (ISA) that determines the status of the graft by simply examining the activation status of blood T cells.
Graft-derived fibroblasts were used as a source of alloantigens and the recipient blood as a source of allograft-specific peripheral blood lymphocytes (PBL). PBL were added to wells containing donor or third-party graft-derived fibroblasts in the presence or absence of interleukin-2 (IL-2). On day 4 [(3)H]thymidine incorporation was quantified after the cells were incubated for 3 days at 37 degrees C, in a 5% CO(2) water-jacketed incubator. The results were analyzed using the following equation: %IL2 - /IL2+ = ((mean[(3)H]thymidine uptake in the absence of IL - 2) / (mean [(3)H]thymidine uptake in the presence of IL - 2)) x 100.
The ISA score (%IL-2 - /IL-2+) correlated strongly with the outcome of the graft, as it had a sensitivity of 82% for detecting rejections (14/17), and a specificity of 81% (30/37) for detecting non-rejections. Notably, the ISA detected immune T cell activation in the blood of graft rejecting subjects, which were not detected by currently used techniques such as mixed lymphocytes reaction.
The ISA is a straightforward procedure that detects allograft rejection with high specificity and sensitivity.
需要一种简单、灵敏、非侵入性的技术来监测移植物功能。我们在此报告一种名为免疫状态检测（ISA）的简单检测方法，该方法通过简单检查血液T细胞的激活状态来确定移植物的状态。
将移植物来源的成纤维细胞用作同种异体抗原的来源，将受体血液用作同种异体移植特异性外周血淋巴细胞（PBL）的来源。在有或无白细胞介素-2（IL-2）的情况下，将PBL添加到含有供体或第三方移植物来源的成纤维细胞的孔中。在第4天，将细胞在37℃、5%二氧化碳的水套式培养箱中孵育3天后，对[³H]胸腺嘧啶核苷掺入量进行定量。使用以下公式分析结果：%IL2 - /IL2+ = （（无IL - 2时的平均[³H]胸腺嘧啶核苷摄取量）/（有IL - 2时的平均[³H]胸腺嘧啶核苷摄取量））×100。
ISA评分（%IL-2 - /IL-2+）与移植物的结果密切相关，其检测排斥反应的灵敏度为82%（14/17），检测非排斥反应的特异性为81%（30/37）。值得注意的是，ISA检测到了移植物排斥受试者血液中的免疫T细胞激活，而目前使用的技术如混合淋巴细胞反应未检测到。
ISA是一种直接的检测方法，具有高特异性和灵敏度来检测同种异体移植排斥反应。
