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Integrin Activation Enables Sensitive Detection of Functional CD4+ and CD8+ T Cells: Application to Characterize SARS-CoV-2 Immunity.
- Source :
- Frontiers in Immunology; 3/29/2021, Vol. 11, pN.PAG-N.PAG, 20p
- Publication Year :
- 2021
-
Abstract
- We have previously shown that conformational change in the β<subscript>2</subscript>-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<superscript>+</superscript> T cells. In this study, we describe a modified protocol of this assay for sensitive detection of functional antigen-specific CD4<superscript>+</superscript> T cells using a monoclonal antibody (clone m24 Ab) specific for the open, high-affinity conformation of the β<subscript>2</subscript>-integrin. The kinetics of β<subscript>2</subscript>-integrin activation was different on CD4<superscript>+</superscript> and CD8<superscript>+</superscript> 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<superscript>+</superscript> and CD8<superscript>+</superscript> 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<subscript>2</subscript>-integrin with m24 and CD154 Abs, we assessed extremely low frequencies of polyfunctional CD4<superscript>+</superscript> T cell responses. The novel assay used in this study allows very sensitive and simultaneous screening of both CD4<superscript>+</superscript> and CD8<superscript>+</superscript> T cell reactivities, with versatile applicability in clinical and vaccination studies. [ABSTRACT FROM AUTHOR]
- Subjects :
- MONONUCLEAR leukocytes
T cells
CELL adhesion molecules
SARS-CoV-2
INTEGRINS
Subjects
Details
- Language :
- English
- ISSN :
- 16643224
- Volume :
- 11
- Database :
- Complementary Index
- Journal :
- Frontiers in Immunology
- Publication Type :
- Academic Journal
- Accession number :
- 149688123
- Full Text :
- https://doi.org/10.3389/fimmu.2021.626308