Soft polydimethylsiloxane-supported lipid bilayers for studying T cell interactions

Lippert, Anna H., Dimov, Ivan B., Winkel, Alexander K., Humphrey, Jane, McColl, James, Chen, Kevin Y., Santos, Ana M., Jenkins, Edward, Franze, Kristian, Davis, Simon J. and Klenerman, David (2021) Soft polydimethylsiloxane-supported lipid bilayers for studying T cell interactions. Biophysical Journal, 120 (1). pp. 35-45. ISSN 0006-3495

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Abstract

Much of what we know about the early stages of T cell activation has been obtained from studies of T cells interacting with glass-supported lipid bilayers that favor imaging but are orders of magnitude stiffer than typical cells. We developed a method for attaching lipid bilayers to polydimethylsiloxane polymer supports, producing "soft bilayers" with physiological levels of mechanical resistance (Young's modulus of 4 kPa). Comparisons of T cell behavior on soft and glass-supported bilayers revealed that whereas late stages of T cell activation are thought to be substrate-stiffness dependent, early calcium signaling was unaffected by substrate rigidity, implying that early steps in T cell receptor triggering are not mechanosensitive. The exclusion of large receptor-type phosphatases was observed on the soft bilayers, however, even though it is yet to be demonstrated at authentic cell-cell contacts. This work sets the stage for an imaging-based exploration of receptor signaling under conditions closely mimicking physiological cell-cell contact.

Item Type: Article
Additional Information: Funding Information: This work was supported by a Research Professorship ( RP150066 to D.K.), the Wellcome Trust ( 098274/Z/12/Z to S.J.D.), the European Research Council (Consolidator Grant 772426 to K.F.), and the Engineering and Physical Sciences Research Council ( EP/L015978/1 to I.B.D). Publisher Copyright: © 2020 Biophysical Society
Uncontrolled Keywords: plasma-membrane,activation,receptor,mobility,tcr,segregation,molecules,adhesion,surface,model,biophysics ,/dk/atira/pure/subjectarea/asjc/1300/1304
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Depositing User: LivePure Connector
Date Deposited: 26 Aug 2022 10:30
Last Modified: 24 Sep 2022 07:02
URI: https://ueaeprints.uea.ac.uk/id/eprint/87638
DOI: 10.1016/j.bpj.2020.11.021

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