|Authors||Fernandez LA, Tsuchida M, Manthei E, Fechner JH, Oberley TD, Leverson GE, Knechtle SJ, Hamawy MM|
|Journal||Transpl. Immunol. Volume: 13 Issue: 2 Pages: 147-54|
|Publish Date||2004 Sep-Oct|
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 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.