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Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity
We have previously shown that conformational change in the β(2)-integrin is a very early activation marker that can be detected with fluorescent multimers of its ligand intercellular adhesion molecule (ICAM)-1 for rapid assessment of antigen-specific CD8(+) T cells. In this study, we describe a modi...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8040333/ https://www.ncbi.nlm.nih.gov/pubmed/33854501 http://dx.doi.org/10.3389/fimmu.2021.626308 |
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author | Schöllhorn, Anna Schuhmacher, Juliane Besedovsky, Luciana Fendel, Rolf Jensen, Anja T. R. Stevanović, Stefan Lange, Tanja Rammensee, Hans-Georg Born, Jan Gouttefangeas, Cécile Dimitrov, Stoyan |
author_facet | Schöllhorn, Anna Schuhmacher, Juliane Besedovsky, Luciana Fendel, Rolf Jensen, Anja T. R. Stevanović, Stefan Lange, Tanja Rammensee, Hans-Georg Born, Jan Gouttefangeas, Cécile Dimitrov, Stoyan |
author_sort | Schöllhorn, Anna |
collection | PubMed |
description | We have previously shown that conformational change in the β(2)-integrin is a very early activation marker that can be detected with fluorescent multimers of its ligand intercellular adhesion molecule (ICAM)-1 for rapid assessment of antigen-specific CD8(+) T cells. In this study, we describe a modified protocol of this assay for sensitive detection of functional antigen-specific CD4(+) T cells using a monoclonal antibody (clone m24 Ab) specific for the open, high-affinity conformation of the β(2)-integrin. The kinetics of β(2)-integrin activation was different on CD4(+) and CD8(+) T cells (several hours vs. few minutes, respectively); however, m24 Ab readily stained both cell types 4–6 h after antigen stimulation. With this protocol, we were able to monitor ex vivo effector and memory CD4(+) and CD8(+) T cells specific for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), cytomegalovirus (CMV), Epstein–Barr virus (EBV), and hepatitis B virus (HBV) in whole blood or cryopreserved peripheral blood mononuclear cells (PBMCs) of infected or vaccinated individuals. By costaining β(2)-integrin with m24 and CD154 Abs, we assessed extremely low frequencies of polyfunctional CD4(+) T cell responses. The novel assay used in this study allows very sensitive and simultaneous screening of both CD4(+) and CD8(+) T cell reactivities, with versatile applicability in clinical and vaccination studies. |
format | Online Article Text |
id | pubmed-8040333 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-80403332021-04-13 Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity Schöllhorn, Anna Schuhmacher, Juliane Besedovsky, Luciana Fendel, Rolf Jensen, Anja T. R. Stevanović, Stefan Lange, Tanja Rammensee, Hans-Georg Born, Jan Gouttefangeas, Cécile Dimitrov, Stoyan Front Immunol Immunology We have previously shown that conformational change in the β(2)-integrin is a very early activation marker that can be detected with fluorescent multimers of its ligand intercellular adhesion molecule (ICAM)-1 for rapid assessment of antigen-specific CD8(+) T cells. In this study, we describe a modified protocol of this assay for sensitive detection of functional antigen-specific CD4(+) T cells using a monoclonal antibody (clone m24 Ab) specific for the open, high-affinity conformation of the β(2)-integrin. The kinetics of β(2)-integrin activation was different on CD4(+) and CD8(+) T cells (several hours vs. few minutes, respectively); however, m24 Ab readily stained both cell types 4–6 h after antigen stimulation. With this protocol, we were able to monitor ex vivo effector and memory CD4(+) and CD8(+) T cells specific for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), cytomegalovirus (CMV), Epstein–Barr virus (EBV), and hepatitis B virus (HBV) in whole blood or cryopreserved peripheral blood mononuclear cells (PBMCs) of infected or vaccinated individuals. By costaining β(2)-integrin with m24 and CD154 Abs, we assessed extremely low frequencies of polyfunctional CD4(+) T cell responses. The novel assay used in this study allows very sensitive and simultaneous screening of both CD4(+) and CD8(+) T cell reactivities, with versatile applicability in clinical and vaccination studies. Frontiers Media S.A. 2021-03-29 /pmc/articles/PMC8040333/ /pubmed/33854501 http://dx.doi.org/10.3389/fimmu.2021.626308 Text en Copyright © 2021 Schöllhorn, Schuhmacher, Besedovsky, Fendel, Jensen, Stevanović, Lange, Rammensee, Born, Gouttefangeas and Dimitrov. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Schöllhorn, Anna Schuhmacher, Juliane Besedovsky, Luciana Fendel, Rolf Jensen, Anja T. R. Stevanović, Stefan Lange, Tanja Rammensee, Hans-Georg Born, Jan Gouttefangeas, Cécile Dimitrov, Stoyan Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title | Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title_full | Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title_fullStr | Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title_full_unstemmed | Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title_short | Integrin Activation Enables Sensitive Detection of Functional CD4(+) and CD8(+) T Cells: Application to Characterize SARS-CoV-2 Immunity |
title_sort | integrin activation enables sensitive detection of functional cd4(+) and cd8(+) t cells: application to characterize sars-cov-2 immunity |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8040333/ https://www.ncbi.nlm.nih.gov/pubmed/33854501 http://dx.doi.org/10.3389/fimmu.2021.626308 |
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