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A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization
T-cell antigen recognition is accompanied by extensive morphological rearrangements of the contact zone between the T-cell and the antigen-presenting cell (APC). This process involves binding of the T-cell receptor (TCR) complex to antigenic peptides presented via MHC on the APC surface, the interac...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7910839/ https://www.ncbi.nlm.nih.gov/pubmed/33504075 http://dx.doi.org/10.3390/cells10020235 |
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author | Fölser, Martin Motsch, Viktoria Platzer, René Huppa, Johannes B. Schütz, Gerhard J. |
author_facet | Fölser, Martin Motsch, Viktoria Platzer, René Huppa, Johannes B. Schütz, Gerhard J. |
author_sort | Fölser, Martin |
collection | PubMed |
description | T-cell antigen recognition is accompanied by extensive morphological rearrangements of the contact zone between the T-cell and the antigen-presenting cell (APC). This process involves binding of the T-cell receptor (TCR) complex to antigenic peptides presented via MHC on the APC surface, the interaction of costimulatory and adhesion proteins, remodeling of the actin cytoskeleton, and the initiation of downstream signaling processes such as the release of intracellular calcium. However, multiparametric time-resolved analysis of these processes is hampered by the difficulty in recording the different readout modalities at high quality in parallel. In this study, we present a platform for simultaneous quantification of TCR distribution via total internal reflection fluorescence microscopy, of intracellular calcium levels, and of T-cell-exerted forces via atomic force microscopy (AFM). In our method, AFM cantilevers were used to bring single T-cells into contact with the activating surface. We designed the platform specifically to enable the study of T-cell triggering via functionalized fluid-supported lipid bilayers, which represent a widely accepted model system to stimulate T-cells in an antigen-specific manner. In this paper, we showcase the possibilities of this platform using primary transgenic T-cells triggered specifically via their cognate antigen presented by MHCII. |
format | Online Article Text |
id | pubmed-7910839 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-79108392021-02-28 A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization Fölser, Martin Motsch, Viktoria Platzer, René Huppa, Johannes B. Schütz, Gerhard J. Cells Article T-cell antigen recognition is accompanied by extensive morphological rearrangements of the contact zone between the T-cell and the antigen-presenting cell (APC). This process involves binding of the T-cell receptor (TCR) complex to antigenic peptides presented via MHC on the APC surface, the interaction of costimulatory and adhesion proteins, remodeling of the actin cytoskeleton, and the initiation of downstream signaling processes such as the release of intracellular calcium. However, multiparametric time-resolved analysis of these processes is hampered by the difficulty in recording the different readout modalities at high quality in parallel. In this study, we present a platform for simultaneous quantification of TCR distribution via total internal reflection fluorescence microscopy, of intracellular calcium levels, and of T-cell-exerted forces via atomic force microscopy (AFM). In our method, AFM cantilevers were used to bring single T-cells into contact with the activating surface. We designed the platform specifically to enable the study of T-cell triggering via functionalized fluid-supported lipid bilayers, which represent a widely accepted model system to stimulate T-cells in an antigen-specific manner. In this paper, we showcase the possibilities of this platform using primary transgenic T-cells triggered specifically via their cognate antigen presented by MHCII. MDPI 2021-01-25 /pmc/articles/PMC7910839/ /pubmed/33504075 http://dx.doi.org/10.3390/cells10020235 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Fölser, Martin Motsch, Viktoria Platzer, René Huppa, Johannes B. Schütz, Gerhard J. A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title | A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title_full | A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title_fullStr | A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title_full_unstemmed | A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title_short | A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization |
title_sort | multimodal platform for simultaneous t-cell imaging, defined activation, and mechanobiological characterization |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7910839/ https://www.ncbi.nlm.nih.gov/pubmed/33504075 http://dx.doi.org/10.3390/cells10020235 |
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