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Related Experiment Video

Updated: Jun 10, 2026

Qualitative and Quantitative Analysis of the Immune Synapse in the Human System Using Imaging Flow Cytometry
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Published on: January 7, 2019

Altered actin centripetal retrograde flow in physically restricted immunological synapses.

Cheng-han Yu1, Hung-jen Wu, Yoshihisa Kaizuka

  • 1Research Centre of Excellence in Mechanobiology, National University of Singapore, Singapore, Singapore.

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|August 6, 2010
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T cell receptor (TCR) clusters move inward with actin retrograde flow during synapse formation. Actin flow velocity decreases toward the synapse center and is affected by TCR clusters, revealing dynamic coupling.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • T cell activation involves the formation of an immunological synapse between T cells and antigen-presenting cells (APCs).
  • The T cell receptor (TCR) and cytoskeletal dynamics, particularly actin, are crucial for synapse assembly and function.
  • Synthetic supported membranes offer a controlled environment to study synapse formation, guiding protein organization via patterned substrates.

Purpose of the Study:

  • To investigate the dynamic relationship between T cell receptor (TCR) clusters and actin flow within the immunological synapse.
  • To analyze how substrate patterns and TCR cluster confinement influence actin dynamics during T cell activation.
  • To elucidate the coupling mechanisms between TCRs and the actin cytoskeleton in a synthetic supported membrane system.

Main Methods:

  • Utilizing mobile anti-CD3epsilon on a spatial-partitioned supported bilayer to ligate and trigger TCR in live Jurkat T cells.
  • Simultaneously tracking T cell receptor (TCR) clusters and GFP-actin speckles using live-cell imaging.
  • Employing flow-based particle tracking algorithms to analyze actin flow fields and velocity distributions across the synapse.

Main Results:

  • Actin retrograde flow was observed to direct the inward transport of TCR clusters.
  • Actin flow velocity decreased towards the center of the synapse.
  • Actin flow velocity was locally reduced by confined TCR clusters but recovered afterward, indicating a dynamic and dissipative coupling.

Conclusions:

  • Actin retrograde flow is a primary driver for the centripetal transport of TCR clusters in the immunological synapse.
  • The interaction between TCR clusters and the actin network is dynamic and dissipative, influencing cytoskeletal flow patterns.
  • This study provides insights into the physical mechanisms governing T cell-APC interactions and synapse formation.