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Related Experiment Videos

Separation of integrin-dependent adhesion from morphological changes based on differential PLC specificities

D K Wooten1, T K Teague, B W McIntyre

  • 1Department of Immunology, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.

Journal of Leukocyte Biology
|January 14, 1999
PubMed
Summary

Lymphocyte activation involves distinct signaling pathways. Phosphatidylinositol-specific phospholipase C (PLC) and protein kinase C (PKC) mediate inside-out signaling, while phosphatidylcholine-specific PLC and PKC handle outside-in signaling for cell adhesion and spreading.

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Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Signaling

Background:

  • Lymphocyte activation involves integrin-mediated adhesion and cell spreading.
  • Protein kinase C (PKC) plays a role in lymphocyte adherence and spreading on fibronectin.
  • Distinct phospholipase C (PLC) inhibitors exhibit different effects on lymphocyte adhesion and morphology.

Purpose of the Study:

  • To elucidate the specific roles of different PLC isoforms and PKC in lymphocyte inside-out and outside-in signaling.
  • To differentiate the signaling pathways governing initial adhesion versus subsequent cell spreading.

Main Methods:

  • Utilized specific inhibitors of phosphatidylinositol-specific PLC (neomycin) and phosphatidylcholine-specific PLC (D609).
  • Investigated the effects of these inhibitors on CD3/CD28-activated T cells and T leukemic cells (HPB-ALL).

Related Experiment Videos

  • Assessed lymphocyte adhesion and cell morphology on fibronectin-coated surfaces.

    • Main Results:

    • Neomycin (phosphatidylinositol-specific PLC inhibitor) blocked spreading by disrupting adhesion in activated T cells.
    • Neomycin did not affect adhesion or morphology when an inside-out signal was not required.
    • D609 (phosphatidylcholine-specific PLC inhibitor) abrogated cell spreading without impacting adhesion.

    Conclusions:

    • Inside-out signaling for integrin alpha4beta1 in lymphocytes involves phosphatidylinositol-specific PLC and PKC.
    • Outside-in signaling for cell spreading utilizes phosphatidylcholine-specific PLC and PKC.
    • These findings delineate distinct molecular mechanisms for integrin activation and ligand-mediated responses in lymphocytes.