Integrin Activation Enables Sensitive Detection of Functional CD4+ and CD8+ T Cells: Application to Characterize SARS-CoV-2 Immunity
| Autor: | 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 |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Front. Immunol. 12:626308 (2021) 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 2021, ' Integrin activation enables sensitive detection of functional CD4 + and CD8 + T cells : Application to characterize SARS-CoV-2 immunity ', Frontiers in Immunology, vol. 12, 626308 . https://doi.org/10.3389/fimmu.2021.626308 |
| ISSN: | 1664-3224 |
| DOI: | 10.3389/fimmu.2021.626308 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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