RT Journal Article
SR Electronic
T1 Probing mechanical interaction of immune receptors and cytoskeleton by membrane nanotube extraction
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.09.15.508080
DO 10.1101/2022.09.15.508080
A1 Fabio Manca
A1 Gautier Eich
A1 Omar N’Dao
A1 Lucie Normand
A1 Kheya Sengupta
A1 Laurent Limozin
A1 Pierre-Henri Puech
YR 2023
UL http://biorxiv.org/content/early/2023/02/23/2022.09.15.508080.abstract
AB 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 StatementThe authors have declared no competing interest.