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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/2/235 |
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| author | Martin Fölser Viktoria Motsch René Platzer Johannes B. Huppa Gerhard J. Schütz |
| author_facet | Martin Fölser Viktoria Motsch René Platzer Johannes B. Huppa Gerhard J. Schütz |
| author_sort | Martin Fölser |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T03:42:58Z |
| format | Article |
| id | doaj.art-f3eecad1365640c599ef7b56ff704913 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T03:42:58Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-f3eecad1365640c599ef7b56ff7049132023-12-03T14:38:18ZengMDPI AGCells2073-44092021-01-0110223510.3390/cells10020235A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological CharacterizationMartin Fölser0Viktoria Motsch1René Platzer2Johannes B. Huppa3Gerhard J. Schütz4Institute of Applied Physics, TU Wien, 1060 Vienna, AustriaInstitute of Applied Physics, TU Wien, 1060 Vienna, AustriaInstitute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, AustriaInstitute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, AustriaInstitute of Applied Physics, TU Wien, 1060 Vienna, AustriaT-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.https://www.mdpi.com/2073-4409/10/2/235T-cellimmunological synapseatomic force microscopytotal internal fluorescence microscopycalcium imaging |
| spellingShingle | Martin Fölser Viktoria Motsch René Platzer Johannes B. Huppa Gerhard J. Schütz A Multimodal Platform for Simultaneous T-cell Imaging, Defined Activation, and Mechanobiological Characterization Cells T-cell immunological synapse atomic force microscopy total internal fluorescence microscopy calcium imaging |
| 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 | T-cell immunological synapse atomic force microscopy total internal fluorescence microscopy calcium imaging |
| url | https://www.mdpi.com/2073-4409/10/2/235 |
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