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Real-time Live Imaging of T-cell Signaling Complex Formation
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Live-cell dynamics and the role of costimulation in immunological synapse formation

Scott A Wetzel1, Timothy W McKeithan, David C Parker

  • 1Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland 97239, USA.

Journal of Immunology (Baltimore, Md. : 1950)
|November 22, 2002
PubMed

Insights

Costimulation is essential for forming mature immunological synapses. Blocking CD80/CD28 or ICAM-1/LFA-1 interactions alters synapse structure and reduces T cell proliferation.

Area of Science:

  • Immunology
  • Cell Biology
  • T cell immunology

Background:

  • The formation of the immunological synapse is critical for T cell activation.
  • Understanding the dynamic processes within the synapse is key to deciphering T cell-APC interactions.

Purpose of the Study:

  • To visualize and analyze the movement of major histocompatibility complex (MHC):peptide complexes during T cell-APC interactions.
  • To investigate the role of costimulatory molecules in the formation and maturation of the immunological synapse.

Main Methods:

  • Utilized live-cell video microscopy of transfected fibroblasts expressing fluorescently tagged MHC:peptide complexes.
  • Monitored antigen recognition via calcium ion concentration changes.
  • Examined the impact of blocking CD80/CD28 and ICAM-1/LFA-1 interactions on synapse formation.

Main Results:

  • Antigen recognition was observed within 30 seconds of T cell-APC contact.
  • Immunological synapses formed within 3-20 minutes, with MHC:peptide complexes accumulating and coalescing.
  • Blocking costimulatory pathways altered synapse morphology, reducing complex accumulation and correlating with decreased T cell proliferation.

Conclusions:

  • Costimulation via CD80/CD28 and ICAM-1/LFA-1 is crucial for the proper formation and maturation of the immunological synapse.
  • While costimulation impacts synapse structure and T cell proliferation, it does not affect early activation markers like CD69, CD25 expression, or T cell receptor (TCR) down-modulation.

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