Activated integrins identify functional antigen-specific CD8+ T cells within minutes after antigen stimulation

Proceedings of the National Academy of Sciences of the United States of America
Stoyan DimitrovHans-Georg Rammensee

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 function...Continue Reading

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Citations

Jul 17, 2019·Cancer Immunology, Immunotherapy : CII·Cécile GouttefangeasStoyan Dimitrov
Feb 14, 2019·The Journal of Experimental Medicine·Stoyan DimitrovLuciana Besedovsky
Jul 23, 2020·Frontiers in Immunology·Valentina BianchiGeorge Coukos
Oct 22, 2019·European Journal of Immunology·Andrea CossarizzaArturo Zychlinsky
Jun 27, 2021·Pediatric Allergy and Immunology : Official Publication of the European Society of Pediatric Allergy and Immunology·Nicola CotugnoPaolo Palma

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Methods Mentioned

BETA
flow cytometry
flow-cytometry
FCS

Software Mentioned

SPSS Statistics
FACS DiVa

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