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Related Experiment Videos

T cell activation via Leu-23 (CD69).

R Testi1, J H Phillips, L L Lanier

  • 1Becton Dickinson Monoclonal Center, Mountain View, CA 94043.

Journal of Immunology (Baltimore, Md. : 1950)
|August 15, 1989
PubMed
Summary

The CD69 activation antigen, a cell surface glycoprotein, initiates T cell proliferation but requires co-stimulation of protein kinase C (PKC). This finding clarifies CD69

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Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • CD69 is an early activation antigen expressed on lymphocytes following activation.
  • Its precise function in T cell signaling and activation remains incompletely understood.
  • CD69 expression is regulated by protein kinase C (PKC) pathways.

Purpose of the Study:

  • To investigate the signaling capacity of the CD69 glycoprotein.
  • To determine if CD69 can independently transmit an activation signal leading to T cell responses.
  • To elucidate the role of CD69 in T cell proliferation and cytokine production.

Main Methods:

  • Flow cytometry to assess CD69 expression.
  • Monoclonal antibody (mAb) cross-linking of CD69 to trigger signaling.
  • Measurement of intracellular calcium ([Ca2+]) levels.
  • Assays for protein kinase C (PKC) activation.
  • Gene expression analysis for IL-2 and IFN-gamma.
  • Assessment of CD25 expression.
  • T cell proliferation assays.
  • Use of phorbol esters (PMA) and Cyclosporin A to modulate signaling pathways.

Main Results:

  • Cross-linking CD69 induced a sustained increase in intracellular calcium, primarily from extracellular influx.
  • CD69 signaling alone did not effectively activate PKC.
  • Combined CD69 stimulation and PKC activation (via PMA) led to IL-2 and IFN-gamma gene expression, CD25 upregulation, and IL-2-dependent T cell proliferation in both CD4+ and CD8+ T cells.
  • Both thymocytes and peripheral T cells required independent PKC activation for proliferation upon CD69 stimulation.
  • Cyclosporin A inhibited CD69-induced cytokine gene expression, despite not affecting PKC-induced CD69 expression.
  • CD69 stimulation did not trigger cytotoxicity programs.

Conclusions:

  • CD69 acts as a costimulatory molecule that, upon engagement, can initiate T cell activation pathways.
  • Effective T cell activation, including cytokine production and proliferation, downstream of CD69 requires concurrent activation of the PKC pathway.
  • CD69 plays a role in regulating T cell responses but is not directly involved in initiating cytotoxic programs.

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