Activation‐induced CD137 is a fast assay for identification and multi‐parameter flow cytometric analysis of alloreactive T cells. (8th September 2013)

• Record Type: Journal Article
• Authors: Litjens, N. H. R.; de Wit, E. A.; Baan, C. C.; Betjes, M. G. H.
• Is Part Of: Clinical and experimental immunology. Volume 174:Number 1(2013:Oct.)
• Page Start: 179
• Page End: 191
• Publication Date: 2013-09-08
• Subjects: alloreactivity -- CD137 -- CD4+ T cells -- CD8+ T cells -- multi‐parameter analysis; Immunopathology -- Periodicals; 616.079
• DOI: 10.1111/cei.12152
• Languages: English
• ISSNs: 0009-9104

Abstract

Summary: Detection and isolation of viable alloreactive T cells at the single‐cell level requires a cell surface marker induced specifically upon T cell receptor activation. In this study, a member of the tumour necrosis factor receptor (TNFR)‐family, CD137 (4‐1BB) was investigated for its potential to identify the total pool of circulating alloreactive T cells. Optimal conditions for sensitive and specific detection of allogeneic‐induced CD137 expression on circulating T cells were established. Thereafter, CD137 + alloreactive T cells were phenotypically and functionally characterized by multi‐parameter flow cytometry. Alloantigen‐induced CD137 expression identified both alloreactive CD8 + T cells (mean ± standard error of the mean: 0·21 ± 0·07%) and alloreactive CD4 + T cells (0·21 ± 0·05%). CD137 + alloreactive T cells were detected in different T cell subsets, including naive T cells, but were found preferentially in CD28 + T cells and not in the terminally differentiated T cell subset. Upon allogeneic (re‐)stimulation, the cytokine‐producing as well as proliferative capacity of T cells resided mainly within the CD137‐expressing fraction. About 10% of the CD137 + alloreactive T cells produced any combination of interferon (IFN)‐γ, interleukin (IL)‐2 and TNF‐α. Polyfunctional alloreactive T cells, defined by multiple cytokine expression, were observed infrequently. In conclusion, activation‐induced CD137 expression is a fast assay allowing for detection and functional analysis of the total alloreactive T cell compartment at the single‐cell level by multi‐parameter flow cytometry.