1. Rapid identification of local T cell expansion in inflammatory organ diseases by flow cytometric T cell receptor Vβ analysis
- Author
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Norbert Sommer, Marc Jacobsen, Antje Necker, Roland Martin, Paolo A. Muraro, Wolfgang H. Oertel, Bernhard Hemmer, James W. Nagle, and Rami Gaber
- Subjects
Adult ,Adolescent ,Receptors, Antigen, T-Cell, alpha-beta ,T-Lymphocytes ,T cell ,Immunology ,Epitopes, T-Lymphocyte ,Streptamer ,Biology ,Lymphocyte Activation ,Interleukin 21 ,Antibody Specificity ,T-Lymphocyte Subsets ,medicine ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,Child ,Antigen-presenting cell ,Aged ,Superantigens ,Infant, Newborn ,Antibodies, Monoclonal ,Infant ,CD28 ,Middle Aged ,Flow Cytometry ,Natural killer T cell ,Clone Cells ,medicine.anatomical_structure ,Child, Preschool ,Acute Disease ,Encephalitis ,CD8 - Abstract
Oligoclonal expansion of antigen-specific T cells occurs frequently during inflammatory diseases. These cells may persist for a long time at high frequency in the body and be enriched in the affected tissues. As a screening test for expanded cell T cell populations at sites of inflammation, we developed an optimized methodology for flow-cytometry-based quantification of T cell receptor Vbeta (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 antibody (mAb) panel recognized approximately two thirds of the T cell receptor alpha/beta repertoire in a group of 64 healthy donors and allowed defining TCR usage in the CD4+ and CD8+ subsets. The reliable detection of expanded Vbeta gene families in T cell populations was confirmed in experiments on superantigen-stimulated T cells. Through differential TCR analysis on T cell subpopulations in cerebrospinal fluid and blood in patients with acute encephalitis, we were able to identify locally expanded CD8+ T cells. The power of this approach affords not only high-throughput comparative TCR analysis for immunological studies in vitro, but also rapid ex vivo identification of cell populations enriched in organ compartments during inflammatory diseases.
- Published
- 2000