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A fast live-cell assay to detect antigen-specific CD8+ T cells based on upregulation of integrin-mediated adhesion
- Source :
- ResearcherID, Publons
- Publication Year :
- 2016
- Publisher :
- The American Association of Immunologists, 2016.
-
Abstract
- Antigen stimulation of T-cell receptors on T cells induces an immediate increase in ligand-binding affinity and clustering in the plasma membrane of beta2-integrins, resulting in a strong binding to intercellular adhesion molecules (ICAMs) on target cells. This firm adhesion, e.g. of cytotoxic T cells to virus-infected or tumor cells with formation of cytotoxic synapses is required for target cell lysis. Therefore, upregulation of integrin-mediated adhesion after stimulation identifies functional effector T cells. Here we present a novel flow cytometry assay to enumerate antigen-specific CD8+ T cells using soluble multimeric ICAM1-Fc-F(ab’)2 fluorescent complexes to identify integrin activation induced after short-term stimulation in vitro. By optimizing and validating this assay we were able to identify pathogen-, tumor- or vaccine-induced CD8+T cells, and show its feasibility for assessing and isolating live, functional antigen-specific CD8+T cells. Performing coexpression analyses, we found that antigen-stimulated cells that expressed cytokines and mobilized CD107a were predominantly highly adherent CD8+T cells. Finally, CD8+ T cells that upregulated integrins were shown to mediate cytolytic activity in an antigen-specific manner. The most striking advantage of this method is the short-time incubation (several minutes) that is necessary to identify and isolate viable antigen-specific cytotoxic T cells by assessing upregulation of T cell adhesiveness, a critical step for elaborating multiple effector functions, including cytotoxicity and cytokine production.
- Subjects :
- Immunology
Immunology and Allergy
Subjects
Details
- ISSN :
- 15506606 and 00221767
- Volume :
- 196
- Database :
- OpenAIRE
- Journal :
- The Journal of Immunology
- Accession number :
- edsair.doi.dedup.....36bb6c2586d1e0f6bb22e64757b768b6