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. 1999 Feb;5(2):117–128.

Suppressive effects of anti-inflammatory agents on human endothelial cell activation and induction of heat shock proteins.

A Amberger 1, M Hala 1, M Saurwein-Teissl 1, B Metzler 1, B Grubeck-Loebenstein 1, Q Xu 1, G Wick 1
PMCID: PMC2230414  PMID: 10203577

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 kappaB (NF-kappaB) 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-alpha (TNF-alpha)-induced adhesion molecule expression on HUVECs, whereas aspirin had an inhibitory effect. These observations correlated with the modulation of NF-kappaB 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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amberger A., Maczek C., Jürgens G., Michaelis D., Schett G., Trieb K., Eberl T., Jindal S., Xu Q., Wick G. Co-expression of ICAM-1, VCAM-1, ELAM-1 and Hsp60 in human arterial and venous endothelial cells in response to cytokines and oxidized low-density lipoproteins. Cell Stress Chaperones. 1997 Jun;2(2):94–103. doi: 10.1379/1466-1268(1997)002<0094:ceoive>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Collins T., Read M. A., Neish A. S., Whitley M. Z., Thanos D., Maniatis T. Transcriptional regulation of endothelial cell adhesion molecules: NF-kappa B and cytokine-inducible enhancers. FASEB J. 1995 Jul;9(10):899–909. [PubMed] [Google Scholar]
  5. Drew A. F., Tipping P. G. Cyclosporine treatment reduces early atherosclerosis in the cholesterol-fed rabbit. Atherosclerosis. 1995 Aug;116(2):181–189. doi: 10.1016/0021-9150(95)05539-9. [DOI] [PubMed] [Google Scholar]
  6. Ellis R. J., van der Vies S. M. Molecular chaperones. Annu Rev Biochem. 1991;60:321–347. doi: 10.1146/annurev.bi.60.070191.001541. [DOI] [PubMed] [Google Scholar]
  7. Emeson E. E., Shen M. L. Accelerated atherosclerosis in hyperlipidemic C57BL/6 mice treated with cyclosporin A. Am J Pathol. 1993 Jun;142(6):1906–1915. [PMC free article] [PubMed] [Google Scholar]
  8. Grubeck-Loebenstein B., Londei M., Greenall C., Pirich K., Kassal H., Waldhäusl W., Feldmann M. Pathogenetic relevance of HLA class II expressing thyroid follicular cells in nontoxic Goiter and in Graves' disease. J Clin Invest. 1988 May;81(5):1608–1614. doi: 10.1172/JCI113495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hansson G. K., Holm J., Jonasson L. Detection of activated T lymphocytes in the human atherosclerotic plaque. Am J Pathol. 1989 Jul;135(1):169–175. [PMC free article] [PubMed] [Google Scholar]
  10. Hansson G. K., Jonasson L., Lojsthed B., Stemme S., Kocher O., Gabbiani G. Localization of T lymphocytes and macrophages in fibrous and complicated human atherosclerotic plaques. Atherosclerosis. 1988 Aug;72(2-3):135–141. doi: 10.1016/0021-9150(88)90074-3. [DOI] [PubMed] [Google Scholar]
  11. Hernández-Pando R., Orozco H., Mancilla R. T-cell lung granulomas induced by sepharose-coupled Mycobacterium tuberculosis protein antigens: immunosuppressive phenomena reversed with cyclophosphamide and indomethacin. Immunology. 1995 Dec;86(4):506–511. [PMC free article] [PubMed] [Google Scholar]
  12. Jurivich D. A., Sistonen L., Kroes R. A., Morimoto R. I. Effect of sodium salicylate on the human heat shock response. Science. 1992 Mar 6;255(5049):1243–1245. doi: 10.1126/science.1546322. [DOI] [PubMed] [Google Scholar]
  13. Kopp E., Ghosh S. Inhibition of NF-kappa B by sodium salicylate and aspirin. Science. 1994 Aug 12;265(5174):956–959. doi: 10.1126/science.8052854. [DOI] [PubMed] [Google Scholar]
  14. Li H., Cybulsky M. I., Gimbrone M. A., Jr, Libby P. Inducible expression of vascular cell adhesion molecule-1 by vascular smooth muscle cells in vitro and within rabbit atheroma. Am J Pathol. 1993 Dec;143(6):1551–1559. [PMC free article] [PubMed] [Google Scholar]
  15. Marcinkiewicz J., Grabowska A., Chain B. M. Is there a role for nitric oxide in regulation of T cell secretion of IL-2? J Immunol. 1996 Jun 15;156(12):4617–4621. [PubMed] [Google Scholar]
  16. Nicoletti I., Migliorati G., Pagliacci M. C., Grignani F., Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271–279. doi: 10.1016/0022-1759(91)90198-o. [DOI] [PubMed] [Google Scholar]
  17. Palmoski M. J., Brandt K. D. Effects of salicylate and indomethacin on glycosaminoglycan and prostaglandin E2 synthesis in intact canine knee cartilage ex vivo. Arthritis Rheum. 1984 Apr;27(4):398–403. doi: 10.1002/art.1780270406. [DOI] [PubMed] [Google Scholar]
  18. Pierce J. W., Read M. A., Ding H., Luscinskas F. W., Collins T. Salicylates inhibit I kappa B-alpha phosphorylation, endothelial-leukocyte adhesion molecule expression, and neutrophil transmigration. J Immunol. 1996 May 15;156(10):3961–3969. [PubMed] [Google Scholar]
  19. Printseva OYu, Peclo M. M., Gown A. M. Various cell types in human atherosclerotic lesions express ICAM-1. Further immunocytochemical and immunochemical studies employing monoclonal antibody 10F3. Am J Pathol. 1992 Apr;140(4):889–896. [PMC free article] [PubMed] [Google Scholar]
  20. Ridker P. M., Cushman M., Stampfer M. J., Tracy R. P., Hennekens C. H. Inflammation, aspirin, and the risk of cardiovascular disease in apparently healthy men. N Engl J Med. 1997 Apr 3;336(14):973–979. doi: 10.1056/NEJM199704033361401. [DOI] [PubMed] [Google Scholar]
  21. Roselaar S. E., Schonfeld G., Daugherty A. Enhanced development of atherosclerosis in cholesterol-fed rabbits by suppression of cell-mediated immunity. J Clin Invest. 1995 Sep;96(3):1389–1394. doi: 10.1172/JCI118174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993 Apr 29;362(6423):801–809. doi: 10.1038/362801a0. [DOI] [PubMed] [Google Scholar]
  23. Schreiber S. L., Crabtree G. R. The mechanism of action of cyclosporin A and FK506. Immunol Today. 1992 Apr;13(4):136–142. doi: 10.1016/0167-5699(92)90111-J. [DOI] [PubMed] [Google Scholar]
  24. Seitz C. S., Kleindienst R., Xu Q., Wick G. Coexpression of heat-shock protein 60 and intercellular-adhesion molecule-1 is related to increased adhesion of monocytes and T cells to aortic endothelium of rats in response to endotoxin. Lab Invest. 1996 Jan;74(1):241–252. [PubMed] [Google Scholar]
  25. Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
  26. Stefanovic H., Filion L. G. Effect of IL-2 and indomethacin on the activation of suppressor cells induced by tetanus toxoid. Immunobiology. 1990 Nov;181(4-5):345–356. doi: 10.1016/S0171-2985(11)80503-6. [DOI] [PubMed] [Google Scholar]
  27. Stemme S., Holm J., Hansson G. K. T lymphocytes in human atherosclerotic plaques are memory cells expressing CD45RO and the integrin VLA-1. Arterioscler Thromb. 1992 Feb;12(2):206–211. doi: 10.1161/01.atv.12.2.206. [DOI] [PubMed] [Google Scholar]
  28. Weber C., Erl W., Pietsch A., Weber P. C. Aspirin inhibits nuclear factor-kappa B mobilization and monocyte adhesion in stimulated human endothelial cells. Circulation. 1995 Apr 1;91(7):1914–1917. doi: 10.1161/01.cir.91.7.1914. [DOI] [PubMed] [Google Scholar]
  29. Welch W. J. Mammalian stress response: cell physiology, structure/function of stress proteins, and implications for medicine and disease. Physiol Rev. 1992 Oct;72(4):1063–1081. doi: 10.1152/physrev.1992.72.4.1063. [DOI] [PubMed] [Google Scholar]
  30. Wick G., Schett G., Amberger A., Kleindienst R., Xu Q. Is atherosclerosis an immunologically mediated disease? Immunol Today. 1995 Jan;16(1):27–33. doi: 10.1016/0167-5699(95)80067-0. [DOI] [PubMed] [Google Scholar]
  31. Xu Q. B., Oberhuber G., Gruschwitz M., Wick G. Immunology of atherosclerosis: cellular composition and major histocompatibility complex class II antigen expression in aortic intima, fatty streaks, and atherosclerotic plaques in young and aged human specimens. Clin Immunol Immunopathol. 1990 Sep;56(3):344–359. doi: 10.1016/0090-1229(90)90155-j. [DOI] [PubMed] [Google Scholar]
  32. Xu Q., Dietrich H., Steiner H. J., Gown A. M., Schoel B., Mikuz G., Kaufmann S. H., Wick G. Induction of arteriosclerosis in normocholesterolemic rabbits by immunization with heat shock protein 65. Arterioscler Thromb. 1992 Jul;12(7):789–799. doi: 10.1161/01.atv.12.7.789. [DOI] [PubMed] [Google Scholar]
  33. Xu Q., Kleindienst R., Waitz W., Dietrich H., Wick G. Increased expression of heat shock protein 65 coincides with a population of infiltrating T lymphocytes in atherosclerotic lesions of rabbits specifically responding to heat shock protein 65. J Clin Invest. 1993 Jun;91(6):2693–2702. doi: 10.1172/JCI116508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Xu Q., Willeit J., Marosi M., Kleindienst R., Oberhollenzer F., Kiechl S., Stulnig T., Luef G., Wick G. Association of serum antibodies to heat-shock protein 65 with carotid atherosclerosis. Lancet. 1993 Jan 30;341(8840):255–259. doi: 10.1016/0140-6736(93)92613-x. [DOI] [PubMed] [Google Scholar]
  35. Zhou X., Stemme S., Hansson G. K. Evidence for a local immune response in atherosclerosis. CD4+ T cells infiltrate lesions of apolipoprotein-E-deficient mice. Am J Pathol. 1996 Aug;149(2):359–366. [PMC free article] [PubMed] [Google Scholar]
  36. Zimarino V., Tsai C., Wu C. Complex modes of heat shock factor activation. Mol Cell Biol. 1990 Feb;10(2):752–759. doi: 10.1128/mcb.10.2.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. van der Wal A. C., Das P. K., Tigges A. J., Becker A. E. Adhesion molecules on the endothelium and mononuclear cells in human atherosclerotic lesions. Am J Pathol. 1992 Dec;141(6):1427–1433. [PMC free article] [PubMed] [Google Scholar]

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