Author:
Dimitrov Stoyan,Gouttefangeas Cécile,Besedovsky Luciana,Jensen Anja T. R.,Chandran P. Anoop,Rusch Elisa,Businger Ramona,Schindler Michael,Lange Tanja,Born Jan,Rammensee Hans-Georg
Abstract
Immediate β2-integrin activation upon T cell receptor stimulation is critical for effective interaction between T cells and their targets and may therefore be used for the rapid identification and isolation of functional T cells. We present a simple and sensitive flow cytometry-based assay to assess antigen-specific T cells using fluorescent intercellular adhesion molecule (ICAM)-1 multimers that specifically bind to activated β2-integrins. The method is compatible with surface and intracellular staining; it is applicable for monitoring of a broad range of virus-, tumor-, and vaccine-specific CD8+ T cells, and for isolating viable antigen-reacting cells. ICAM-1 binding correlates with peptide-MHC multimer binding but, notably, it identifies the fraction of antigen-specific CD8+ T cells with immediate and high functional capability (i.e., expressing high levels of cytotoxic markers and cytokines). Compared with the currently available methods, staining of activated β2-integrins presents the unique advantage of requiring activation times of only several minutes, therefore delivering functional information nearly reflecting the in vivo situation. Hence, the ICAM-1 assay is most suitable for rapid and precise monitoring of functional antigen-specific T cell responses, including for patient samples in a variety of clinical settings, as well as for the isolation of functional T cells for adoptive cell-transfer immunotherapies.
Funder
Deutsche Forschungsgemeinschaft
Bundesministerium für Bildung und Forschung
ERC advanced grant
Publisher
Proceedings of the National Academy of Sciences
Cited by
19 articles.
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