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Interferon-gamma-induced epithelial ICAM-1 expression and monocyte adhesion. Involvement of protein kinase

Ya-Jen Chang1, Michael J Holtzman, Ching-Chow Chen

  • 1Department of Pharmacology, College of Medicine, National Taiwan University, No. 1 Jen-Ai Road, 1st Section, Taipei 10018, Taiwan.

The Journal of Biological Chemistry
|December 26, 2001
PubMed

Insights

Interferon-gamma (IFN-gamma) increases ICAM-1 expression and U937 cell adhesion in NCI-H292 cells. This pathway involves tyrosine kinases, PLC-gamma2, PKC-alpha, c-Src, STAT1alpha, and the GAS element in the ICAM-1 promoter.

Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • Intercellular adhesion molecule-1 (ICAM-1) plays a crucial role in immune cell interactions.
  • Interferon-gamma (IFN-gamma) is a key cytokine involved in immune responses.
  • Understanding the signaling pathways regulating ICAM-1 expression is vital for immune modulation.

Purpose of the Study:

  • To elucidate the molecular mechanisms by which IFN-gamma induces ICAM-1 expression in NCI-H292 epithelial cells.
  • To identify the key signaling molecules and pathways involved in IFN-gamma-mediated ICAM-1 upregulation.
  • To investigate the role of tyrosine kinases, phospholipase C, and protein kinase C in this process.

Main Methods:

  • Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence staining to assess ICAM-1 expression.
  • Use of specific inhibitors for tyrosine kinases, Src family, phosphatidylinositol-phospholipase C (PLC), and protein kinase C (PKC).
  • Reporter gene assays (luciferase) with wild-type and mutant ICAM-1 promoter constructs, electrophoretic gel mobility shift assays (EMSA), and kinase assays.

Main Results:

  • IFN-gamma significantly increased ICAM-1 expression and U937 cell adhesion to NCI-H292 cells.
  • Tyrosine kinase inhibitors, Src inhibitor PP2, PLC inhibitor U73122, and PKC inhibitors (staurosporine, Ro 31-8220) attenuated IFN-gamma-induced ICAM-1 expression.
  • IFN-gamma and TPA (PKC activator) enhanced ICAM-1 promoter activity via GAS and NF-kappaB sites, involving STAT1alpha activation. Both IFN-gamma and TPA activated c-Src and Lyn kinases.
  • Cotransfection studies indicated that PLC-gamma2, PKCalpha, c-Src, JAK1, and STAT1 are essential components of the signaling cascade.

Conclusions:

  • IFN-gamma induces ICAM-1 expression and subsequent monocyte adhesion in NCI-H292 cells through a signaling pathway involving tyrosine kinases, PLC-gamma2, PKC-alpha, c-Src/Lyn, STAT1alpha, and the GAS element of the ICAM-1 promoter.
  • PKC activation is downstream of tyrosine kinase and PLC-gamma2 activation.
  • This study elucidates a critical signaling cascade regulating ICAM-1 expression in epithelial cells, relevant to inflammatory and immune responses.

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