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Suppressive Effects of Anti-inflammatory Agents on Human Endothelial Cell Activation and Induction of Heat Shock Proteins

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Abstract

Background

Studies from our laboratory have shown that the earliest stages of atherosclerosis may be mediated by an autoimmune reaction against heat shock protein 60 (Hsp60). The interactions of Hsp60-specific T cells with arterial endothelial cells (EC) require expression of both Hsp60 and certain adhesion molecules shown to be induced simultaneously in EC by mechanical and other types of stress. Recently, it was shown that suppression of T cell-mediated immune responses by cyclosporin A (CyA) enhanced atherosclerotic lesion formation in mice. In contrast, aspirin was found to lower the risk of myocardial infarction in men. These conflicting observations may be due to different effects of anti-inflammatory agents on adhesion molecule and Hsp expression in EC, respectively.

Material and Methods

In the present study, we analyzed the effects of CyA, aspirin, and indomethacin on T cell proliferation using a proliferation assay. To explore the expression of adhesion molecules, monocyte chemoattractant protein-1 (MCP-1), and Hsp60 in human umbilical vein endothelial cells (HUVECs), Northern blot analyses were used. To examine the activation status of the transcription factors nuclear factor κB (NF-κB) and heat shock factor-1 (HSF-1), electrophoretic mobility shift assays were performed.

Results

With the exception of indomethacin, the used immunosuppressive and anti-inflammatory agents significantly inhibited T cell proliferation in response to influenza virus antigen in a dose-dependent manner. Interestingly, CyA and indomethacin did not suppress tumor necrosis factor-α (TNF-α)-induced adhesion molecule expression on HUVECs, whereas aspirin had an inhibitory effect. These observations correlated with the modulation of NF-κB activity in EC. All agents tested induced expression of Hsp60 6 hr after application. In addition, aspirin and indomethacin, but not CyA, induced Hsp70 expression in HUVECs that correlated with induction of HSF-1 activity.

Conclusion

Our results show that the tested agents (except indomethacin) are inhibitors of the T cell-mediated immune response, as expected, that aspirin is an effective suppressor of adhesion molecule expression, and that all three agents can induce Hsp60 in HUVECs. These data provide the molecular basis for the notion that (1) part of the anti-atherogenic effect of aspirin may be due to the prevention of the adhesion of sensitized T cells to stressed EC; (2) that part of the atherosclerosis-promoting effect of CyA may be due to its potential as an inducer of Hsp60 expression and its inability to down-regulate adhesion molecule expression on EC; and (3) that down-regulation of MCP-1 expression by aspirin may result in decreased recruitment of monocytes into the arterial intima beneath stressed EC.

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Acknowledgements

This work was supported by the Austrian Research Council Project 12715 (G. W.) and the Sandoz Foundation for Gerontological Research (G. W.) The authors thank G. Sturm for her excellent technical assistance and T. Öttl for his help in preparation of the photographs.

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Authors and Affiliations

  1. Institute for Biomedical Aging Research, Austrian Academy of Sciences, Rennweg 10, A-6020, Innsbruck, Austria

    Albert Amberger, Monika Hala, Maria Saurwein-Teissl, Bernhard Metzler, Beatrix Grubeck-Loebenstein, Qingbo Xu & Georg Wick M.D.

  2. Institute for General and Experimental Pathology, Medical School, University of Innsbruck, Innsbruck, Austria

    Georg Wick M.D.

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  1. Albert Amberger
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  2. Monika Hala
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  3. Maria Saurwein-Teissl
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Correspondence to Georg Wick M.D..

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Amberger, A., Hala, M., Saurwein-Teissl, M. et al. Suppressive Effects of Anti-inflammatory Agents on Human Endothelial Cell Activation and Induction of Heat Shock Proteins. Mol Med 5, 117–128 (1999). https://doi.org/10.1007/BF03402146

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