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Controlling T cells spreading, mechanics and activation by micropatterning
We designed a strategy, based on a careful examination of the activation capabilities of proteins and antibodies used as substrates for adhering T cells, coupled to protein microstamping to control at the same time the position, shape, spreading, mechanics and activation state of T cells. Once adher...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7991639/ https://www.ncbi.nlm.nih.gov/pubmed/33762632 http://dx.doi.org/10.1038/s41598-021-86133-1 |
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author | Sadoun, Anaïs Biarnes-Pelicot, Martine Ghesquiere-Dierickx, Laura Wu, Ambroise Théodoly, Olivier Limozin, Laurent Hamon, Yannick Puech, Pierre-Henri |
author_facet | Sadoun, Anaïs Biarnes-Pelicot, Martine Ghesquiere-Dierickx, Laura Wu, Ambroise Théodoly, Olivier Limozin, Laurent Hamon, Yannick Puech, Pierre-Henri |
author_sort | Sadoun, Anaïs |
collection | PubMed |
description | We designed a strategy, based on a careful examination of the activation capabilities of proteins and antibodies used as substrates for adhering T cells, coupled to protein microstamping to control at the same time the position, shape, spreading, mechanics and activation state of T cells. Once adhered on patterns, we examined the capacities of T cells to be activated with soluble anti CD3, in comparison to T cells adhered to a continuously decorated substrate with the same density of ligands. We show that, in our hand, adhering onto an anti CD45 antibody decorated surface was not affecting T cell calcium fluxes, even adhered on variable size micro-patterns. Aside, we analyzed the T cell mechanics, when spread on pattern or not, using Atomic Force Microscopy indentation. By expressing MEGF10 as a non immune adhesion receptor in T cells we measured the very same spreading area on PLL substrates and Young modulus than non modified cells, immobilized on anti CD45 antibodies, while retaining similar activation capabilities using soluble anti CD3 antibodies or through model APC contacts. We propose that our system is a way to test activation or anergy of T cells with defined adhesion and mechanical characteristics, and may allow to dissect fine details of these mechanisms since it allows to observe homogenized populations in standardized T cell activation assays. |
format | Online Article Text |
id | pubmed-7991639 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-79916392021-03-26 Controlling T cells spreading, mechanics and activation by micropatterning Sadoun, Anaïs Biarnes-Pelicot, Martine Ghesquiere-Dierickx, Laura Wu, Ambroise Théodoly, Olivier Limozin, Laurent Hamon, Yannick Puech, Pierre-Henri Sci Rep Article We designed a strategy, based on a careful examination of the activation capabilities of proteins and antibodies used as substrates for adhering T cells, coupled to protein microstamping to control at the same time the position, shape, spreading, mechanics and activation state of T cells. Once adhered on patterns, we examined the capacities of T cells to be activated with soluble anti CD3, in comparison to T cells adhered to a continuously decorated substrate with the same density of ligands. We show that, in our hand, adhering onto an anti CD45 antibody decorated surface was not affecting T cell calcium fluxes, even adhered on variable size micro-patterns. Aside, we analyzed the T cell mechanics, when spread on pattern or not, using Atomic Force Microscopy indentation. By expressing MEGF10 as a non immune adhesion receptor in T cells we measured the very same spreading area on PLL substrates and Young modulus than non modified cells, immobilized on anti CD45 antibodies, while retaining similar activation capabilities using soluble anti CD3 antibodies or through model APC contacts. We propose that our system is a way to test activation or anergy of T cells with defined adhesion and mechanical characteristics, and may allow to dissect fine details of these mechanisms since it allows to observe homogenized populations in standardized T cell activation assays. Nature Publishing Group UK 2021-03-24 /pmc/articles/PMC7991639/ /pubmed/33762632 http://dx.doi.org/10.1038/s41598-021-86133-1 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Sadoun, Anaïs Biarnes-Pelicot, Martine Ghesquiere-Dierickx, Laura Wu, Ambroise Théodoly, Olivier Limozin, Laurent Hamon, Yannick Puech, Pierre-Henri Controlling T cells spreading, mechanics and activation by micropatterning |
title | Controlling T cells spreading, mechanics and activation by micropatterning |
title_full | Controlling T cells spreading, mechanics and activation by micropatterning |
title_fullStr | Controlling T cells spreading, mechanics and activation by micropatterning |
title_full_unstemmed | Controlling T cells spreading, mechanics and activation by micropatterning |
title_short | Controlling T cells spreading, mechanics and activation by micropatterning |
title_sort | controlling t cells spreading, mechanics and activation by micropatterning |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7991639/ https://www.ncbi.nlm.nih.gov/pubmed/33762632 http://dx.doi.org/10.1038/s41598-021-86133-1 |
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