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Schöllhorn, A.* ; Schuhmacher, J.* ; Besedovsky, L.* ; Fendel, R.* ; Jensen, A.T.R.* ; Stevanović, S.* ; Lange, T.* ; Rammensee, H.G.* ; Born, J. ; Gouttefangeas, C.* ; Dimitrov, S.I.*

Integrin activation enables sensitive detection of functional CD4+ and CD8+ T cells: Application to characterize SARS-CoV-2 immunity.

Front. Immunol. 12:626308 (2021)
Publ. Version/Full Text Research data DOI PMC
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We have previously shown that conformational change in the β2-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 β2-integrin. The kinetics of β2-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 β2-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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Cd4+ T Cells ; Cd8+ T Cells ; Sars-cov-2 ; Antigen Specificity ; Integrin Activation
Language english
Publication Year 2021
HGF-reported in Year 2021
ISSN (print) / ISBN 1664-3224
e-ISSN 1664-3224
Journal Frontiers in Immunology
Quellenangaben Volume: 12, Issue: , Pages: , Article Number: 626308 Supplement: ,
Publisher Frontiers
Publishing Place Avenue Du Tribunal Federal 34, Lausanne, Ch-1015, Switzerland
Reviewing status Peer reviewed
Institute(s) Institute of Diabetes Research and Metabolic Diseases (IDM)
POF-Topic(s) 90000 - German Center for Diabetes Research
Research field(s) Helmholtz Diabetes Center
PSP Element(s) G-502400-001
Grants Deutsche Forschungsgemeinschaft
Deutsche Forschungsgemeinschaft under Germany's Excellence Strategy
Deutsche Forschungsgemeinschaft, Collaborative Research Center
DOI 10.3389/fimmu.2021.626308
WOS ID WOS:000639244300001
Scopus ID 85104137366
PubMed ID 33854501
Erfassungsdatum 2021-04-22
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