Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6

Joost te Riet, Jonne Helenius, Nico Strohmeyer, Alessandra Cambi, Carl Figdor, Daniel J. Müller

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Abstract

At the immunological synapse, the activated leukocyte cell adhesion molecule (ALCAM) on a dendritic cell (DC) and CD6 molecules on a T cell contribute to sustained DC–T-cell contacts. However, little is known about how ALCAM–CD6 bonds resist and adapt to mechanical stress. Here, we combine single-cell force spectroscopy (SCFS) with total-internal reflection fluorescence microscopy to examine ALCAM–CD6-mediated cell adhesion. The combination of cells expressing ALCAM constructs with certain cytoplasmic tail mutations and improved SCFS analysis processes reveal that the affinity of ALCAM–CD6 bonds is not influenced by the linking of the intracellular domains of ALCAM to the actin cortex. By contrast, the recruitment of ALCAM to adhesion sites and the propensity of ALCAM to anchor plasma membrane tethers depend on actin cytoskeletal interactions. Furthermore, linking ALCAM to the actin cortex through adaptor proteins stiffens the cortex and strengthens cell adhesion. We propose a framework for how ALCAMs contribute to DC–T-cell adhesion, stabilize DC–T-cell contacts and form a mechanical link between CD6 and the actin cortex to strengthen cell adhesion at the immunological synapse
Original languageEnglish
Pages (from-to)1595-1606
Number of pages12
JournalJournal of cell science
Volume127
Issue number7
DOIs
Publication statusPublished - 2014

Keywords

  • Single-cell force spectroscopy
  • Atomic force microscopy
  • ALCAM
  • CD6
  • Cell adhesion
  • Immunological synapse
  • Membrane tethers

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    te Riet, J., Helenius, J., Strohmeyer, N., Cambi, A., Figdor, C., & Müller, D. J. (2014). Dynamic coupling of ALCAM to the actin cortex strengthens cell adhesion to CD6. Journal of cell science, 127(7), 1595-1606. https://doi.org/10.1242/jcs.141077