Rapid identification of local T cell expansion in inflammatory organ diseases by flow cytometric T cell receptor V beta analysis

Oligoclonal expansion of antigen-specific T cells occurs frequently during inflammatory diseases. These cells may persist for a long time at high freq uency in the body and be enriched in the affected tissues. As a screening t est for expanded cell T cell populations at sites of inflammation, we devel oped an optimized methodology for flow-cytometry-based quantification of T cell receptor V beta (TCRBV) expression. We first validated the specificity of a TCRBV-specific monoclonal antibody set by direct comparison with PCR- based analysis of mono- and polyclonal T cell samples. This monoclonal anti body (mAb) panel recognized approximately two thirds of the T cell receptor alpha/beta repertoire in agroup of 64 healthy donors and allowed defining TCR usage in the CD4+ and CD8+ subsets. The reliable detection of expanded V beta gene families in T cell populations was confirmed in experiments on superantigen-stimulated T cells. Through differential TCR analysis on T cel l subpopulations in cerebrospinal fluid and blood in patients with acute en cephalitis, we were able to identify locally expanded CD8+ T cells. The pow er of this approach affords not only high-throughput comparative TCR analys is for immunological studies in vitro, but also rapid ex vivo identificatio n of cell populations enriched in organ compartments during inflammatory di seases. (C) 2000 Elsevier Science B.V; All rights reserved.