Soft polydimethylsiloxane-supported lipid bilayers for studying T cell interactions

Anna H. Lippert, Ivan B. Dimov, Alexander K. Winkel, Jane Humphrey, James McColl, Kevin Y. Chen, Ana M. Santos, Edward Jenkins, Kristian Franze, Simon J. Davis, David Klenerman

Research output: Contribution to journalArticlepeer-review

4 Citations (SciVal)

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.

Original languageEnglish
Pages (from-to)35-45
Number of pages11
JournalBiophysical Journal
Volume120
Issue number1
Early online date26 Nov 2020
DOIs
Publication statusPublished - 5 Jan 2021
Externally publishedYes

Keywords

  • PLASMA-MEMBRANE
  • ACTIVATION
  • RECEPTOR
  • MOBILITY
  • TCR
  • SEGREGATION
  • MOLECULES
  • ADHESION
  • SURFACE
  • MODEL

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