Micropatterning of TCR and LFA-1 ligands reveals complementary effects on cytoskeleton mechanics in T cells

Integr Biol (Camb). 2015 Oct;7(10):1272-84. doi: 10.1039/c5ib00032g. Epub 2015 Jul 9.

Abstract

The formation of the immunological synapse between a T cell and the antigen-presenting cell (APC) is critically dependent on actin dynamics, downstream of T cell receptor (TCR) and integrin (LFA-1) signalling. There is also accumulating evidence that mechanical forces, generated by actin polymerization and/or myosin contractility regulate T cell signalling. Because both receptor pathways are intertwined, their contributions towards the cytoskeletal organization remain elusive. Here, we identify the specific roles of TCR and LFA-1 by using a combination of micropatterning to spatially separate signalling systems and nanopillar arrays for high-precision analysis of cellular forces. We identify that Arp2/3 acts downstream of TCRs to nucleate dense actin foci but propagation of the network requires LFA-1 and the formin FHOD1. LFA-1 adhesion enhances actomyosin forces, which in turn modulate actin assembly downstream of the TCR. Together our data shows a mechanically cooperative system through which ligands presented by an APC modulate T cell activation.

MeSH terms

  • Actins / physiology
  • Antigen Presentation
  • Antigen-Presenting Cells / immunology
  • Biomechanical Phenomena
  • CD4-Positive T-Lymphocytes / immunology*
  • CD4-Positive T-Lymphocytes / physiology*
  • Cell Adhesion
  • Cells, Cultured
  • Cytoskeleton / physiology*
  • Humans
  • Immunological Synapses / physiology
  • Ligands
  • Lymphocyte Activation
  • Lymphocyte Function-Associated Antigen-1 / metabolism*
  • Models, Immunological
  • Receptors, Antigen, T-Cell / metabolism*
  • Signal Transduction

Substances

  • Actins
  • Ligands
  • Lymphocyte Function-Associated Antigen-1
  • Receptors, Antigen, T-Cell