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Probing mechanical interaction of immune receptors and cytoskeleton by membrane nanotube extraction

View ORCID ProfileFabio Manca, Gautier Eich, Omar N’Dao, Lucie Normand, View ORCID ProfileKheya Sengupta, View ORCID ProfileLaurent Limozin, View ORCID ProfilePierre-Henri Puech
doi: https://doi.org/10.1101/2022.09.15.508080
Fabio Manca
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
bAix Marseille University, CNRS, Centre Interdisciplinaire de Nanoscience de Marseille (CINaM), 13009 Marseille, France
cTuring Center for Living Systems (CENTURI), 13009 Marseille, France
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Gautier Eich
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
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Omar N’Dao
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
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Lucie Normand
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
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Kheya Sengupta
bAix Marseille University, CNRS, Centre Interdisciplinaire de Nanoscience de Marseille (CINaM), 13009 Marseille, France
cTuring Center for Living Systems (CENTURI), 13009 Marseille, France
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Laurent Limozin
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
cTuring Center for Living Systems (CENTURI), 13009 Marseille, France
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Pierre-Henri Puech
aAix Marseille University, CNRS, INSERM, Laboratory Adhesion and Inflammation (LAI), 13009 Marseille, France
cTuring Center for Living Systems (CENTURI), 13009 Marseille, France
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  • For correspondence: pierre-henri.puech@inserm.fr
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Abstract

The role of force application in immune cell recognition is now well established, the force being transmitted between the actin cytoskeleton to the anchoring ligands through receptors such as integrins. In this chain, the mechanics of the cytoskeleton to receptor link, though clearly crucial, remains poorly understood. To probe this link, we combine mechanical extraction of membrane tubes from T cells using optical tweezers, and fitting of the resulting force curves with a viscoelastic model taking into account the cell and relevant molecules. We solicit this link using four different antibodies against various membrane bound receptors: antiCD3 to target the T Cell Receptor (TCR) complex, antiCD45 for the long sugar CD45, and two clones of antiCD11 targeting open or closed conformation of LFA1 integrins. Upon disruption of the cytoskeleton, the stiffness of the link changes for two of the receptors, exposing the existence of a receptor to cytoskeleton link - namely TCR-complex and open LFA1, and does not change for the other two where no such a link was expected. Our integrated approach allows us to probe, for the first time, the mechanics of the intracellular receptor-cytoskeleton link in immune cells.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵1 Emails: fabio.manca{at}inserm.fr

  • ↵2 Emails: kheya.sengupta{at}cnrs.fr

  • ↵3 Emails: laurent.limozin{at}inserm.fr

  • The authors declare no conflict of interest.

  • - introduction has been improved - discussion has been improved - addition of a new supporting figure for discussion - addition of new references

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 23, 2023.
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Probing mechanical interaction of immune receptors and cytoskeleton by membrane nanotube extraction
Fabio Manca, Gautier Eich, Omar N’Dao, Lucie Normand, Kheya Sengupta, Laurent Limozin, Pierre-Henri Puech
bioRxiv 2022.09.15.508080; doi: https://doi.org/10.1101/2022.09.15.508080
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Probing mechanical interaction of immune receptors and cytoskeleton by membrane nanotube extraction
Fabio Manca, Gautier Eich, Omar N’Dao, Lucie Normand, Kheya Sengupta, Laurent Limozin, Pierre-Henri Puech
bioRxiv 2022.09.15.508080; doi: https://doi.org/10.1101/2022.09.15.508080

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