Immune status assay (ISA): a noninvasive procedure for studying allograft rejection.

Journal Article (Journal Article)

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: The ISA is a straightforward procedure that detects allograft rejection with high specificity and sensitivity.

Full Text

Duke Authors

Cited Authors

  • Fernandez, LA; Tsuchida, M; Manthei, E; Fechner, JH; Oberley, TD; Leverson, GE; Knechtle, SJ; Hamawy, MM

Published Date

  • September 2004

Published In

Volume / Issue

  • 13 / 2

Start / End Page

  • 147 - 154

PubMed ID

  • 15380545

International Standard Serial Number (ISSN)

  • 0966-3274

Digital Object Identifier (DOI)

  • 10.1016/j.trim.2004.05.007


  • eng

Conference Location

  • Netherlands