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CD154-stimulated GM-CSF release by vascular smooth muscle cells elicits monocyte activation—role in atherogenesis

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Abstract

During the early phase of atherosclerosis, T cells and monocytes attach to and migrate through the endothelium into the vessel wall. To provide an insight into the potential cross talk between T cells and smooth muscle cells (SMC) in atherogenesis, we investigated changes in gene expression caused by CD40 ligation in cultured vascular SMC and their consequences for monocyte activation. CD40 expression in human-cultured SMC was induced by 24-h treatment with tumor necrosis factor-α plus interferon-γ followed by 12-h exposure to mouse myeloma cells stably expressing human CD154 or the corresponding control cells. DNA microarray analysis (Affymetrix HG-U952A chip) indicated 33 up-regulated genes in three individual experiments of which 19 encoded pro-inflammatory adhesion molecules, cytokines, chemokines, and receptors. One functional consequence of this change in gene expression was an activation of transformed human promonocytic-1 monocytes exposed to the conditioned medium of the stimulated SMC. Subsequent antibody neutralization experiments identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as the SMC-derived cytokine responsible for this effect. Thus, vascular SMC-like endothelial cells appear to contribute to the maintenance of an inflammatory response in the atherosclerotic vessel wall upon CD40–CD154 co-stimulation. Among 19 up-regulated pro-inflammatory gene products, GM-CSF plays an important role in SMC-dependent monocyte activation.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (SFB 405/B17). The expert technical assistance of Annette Benemann and Simone Pöschel is gratefully acknowledged.

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Authors and Affiliations

  1. Institute of Physiology and Pathophysiology, Division of Cardiovascular Physiology, University Hospital Heidelberg, Im Neuenheimer Feld 326, 69120, Heidelberg, Germany

    Milica Stojakovic, Robert Krzesz, Andreas H. Wagner & Markus Hecker

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  1. Milica Stojakovic
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  2. Robert Krzesz
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  3. Andreas H. Wagner
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  4. Markus Hecker
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Corresponding author

Correspondence to Markus Hecker.

Additional information

Milica Stojakovic and Robert Krzesz contributed equally to this work.

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Supplementary Table 1

Primer sequences and PCR conditions. Asterisk real-time PCR primers). CCR7 Chemokine CC motif receptor 7, GAPDH glyceraldehyde-3-phophate dehydrogenase, MIP-2β macrophage inflammatory protein-2β, IL-1β interleukin-1β, MCP-1 monocyte chemoattractant protein 1, MMP-3 metalloproteinase 3, RPL32 ribosomal protein L32, VCAM-1 vascular cell adhesion molecule-1. Primer pairs used were specific for the human variant of the listed genes (PDF 85 kb).

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Stojakovic, M., Krzesz, R., Wagner, A.H. et al. CD154-stimulated GM-CSF release by vascular smooth muscle cells elicits monocyte activation—role in atherogenesis. J Mol Med 85, 1229–1238 (2007). https://doi.org/10.1007/s00109-007-0225-y

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  • DOI: https://doi.org/10.1007/s00109-007-0225-y

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