Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Apr 1;97(7):1715–1722. doi: 10.1172/JCI118598

Activated transcription factor nuclear factor-kappa B is present in the atherosclerotic lesion.

K Brand 1, S Page 1, G Rogler 1, A Bartsch 1, R Brandl 1, R Knuechel 1, M Page 1, C Kaltschmidt 1, P A Baeuerle 1, D Neumeier 1
PMCID: PMC507236  PMID: 8601637

Abstract

Nuclear factor-kappa B (NF-kappaB)/Rel transcription factors play an important role in the inducible regulation of a variety of genes involved in the inflammatory and proliferative responses of cells. The present study was designed to elucidate the implication of NF-kappaB/Rel in the pathogenesis of atherosclerosis. Activation of the dimeric NF-kappaB complex is regulated at a posttranslational level and requires the release of the inhibitor protein IkappaB. The newly developed mAb alpha-p65mAb recognizes the IkappaB binding region on the p65 (RelA) DNA binding subunit and therefore selectively reacts with p65 in activated NF-kappaB. Using immunofluorescence and immunohistochemical techniques, activated NF-kappaB was detected in the fibrotic-thickened intima/media and atheromatous areas of the atherosclerotic lesion. Activation of NF-kappaB was identified in smooth muscle cells, macrophages, and endothelial cells. Little or no activated NF-kappaB was detected in vessels lacking atherosclerosis. Electrophoretic mobility shift assays and colocalization of activated NF-kappaB with NF-kappaB target gene expression suggest functional implications for this transcription factor in the atherosclerotic lesion. This study demonstrates the presence of activated NF-kappaB in human atherosclerotic tissue for the first time. Atherosclerosis, characterized by features of chronic inflammation and proliferative processes, may be a paradigm for the involvement of NF-kappaB/Rel in chronic inflammatory disease.

Full Text

The Full Text of this article is available as a PDF (521.6 KB).

Selected References

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

  1. Amento E. P., Ehsani N., Palmer H., Libby P. Cytokines and growth factors positively and negatively regulate interstitial collagen gene expression in human vascular smooth muscle cells. Arterioscler Thromb. 1991 Sep-Oct;11(5):1223–1230. doi: 10.1161/01.atv.11.5.1223. [DOI] [PubMed] [Google Scholar]
  2. Badimon J. J., Fuster V., Chesebro J. H., Badimon L. Coronary atherosclerosis. A multifactorial disease. Circulation. 1993 Mar;87(3 Suppl):II3–I16. [PubMed] [Google Scholar]
  3. Baeuerle P. A., Baltimore D. I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science. 1988 Oct 28;242(4878):540–546. doi: 10.1126/science.3140380. [DOI] [PubMed] [Google Scholar]
  4. Baeuerle P. A., Henkel T. Function and activation of NF-kappa B in the immune system. Annu Rev Immunol. 1994;12:141–179. doi: 10.1146/annurev.iy.12.040194.001041. [DOI] [PubMed] [Google Scholar]
  5. Baeuerle P. A. The inducible transcription activator NF-kappa B: regulation by distinct protein subunits. Biochim Biophys Acta. 1991 Apr 16;1072(1):63–80. doi: 10.1016/0304-419x(91)90007-8. [DOI] [PubMed] [Google Scholar]
  6. Barath P., Fishbein M. C., Cao J., Berenson J., Helfant R. H., Forrester J. S. Tumor necrosis factor gene expression in human vascular intimal smooth muscle cells detected by in situ hybridization. Am J Pathol. 1990 Sep;137(3):503–509. [PMC free article] [PubMed] [Google Scholar]
  7. Beutler B., Van Huffel C. An evolutionary and functional approach to the TNF receptor/ligand family. Ann N Y Acad Sci. 1994 Aug 15;730:118–133. doi: 10.1111/j.1749-6632.1994.tb44244.x. [DOI] [PubMed] [Google Scholar]
  8. Brand K., Banka C. L., Mackman N., Terkeltaub R. A., Fan S. T., Curtiss L. K. Oxidized LDL enhances lipopolysaccharide-induced tissue factor expression in human adherent monocytes. Arterioscler Thromb. 1994 May;14(5):790–797. doi: 10.1161/01.atv.14.5.790. [DOI] [PubMed] [Google Scholar]
  9. Brand K., Fowler B. J., Edgington T. S., Mackman N. Tissue factor mRNA in THP-1 monocytic cells is regulated at both transcriptional and posttranscriptional levels in response to lipopolysaccharide. Mol Cell Biol. 1991 Sep;11(9):4732–4738. doi: 10.1128/mcb.11.9.4732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Brand K., Mackman N., Curtiss L. K. Interferon-gamma inhibits macrophage apolipoprotein E production by posttranslational mechanisms. J Clin Invest. 1993 May;91(5):2031–2039. doi: 10.1172/JCI116425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cheng Q., Cant C. A., Moll T., Hofer-Warbinek R., Wagner E., Birnstiel M. L., Bach F. H., de Martin R. NK-kappa B subunit-specific regulation of the I kappa B alpha promoter. J Biol Chem. 1994 May 6;269(18):13551–13557. [PubMed] [Google Scholar]
  12. Clinton S. K., Underwood R., Hayes L., Sherman M. L., Kufe D. W., Libby P. Macrophage colony-stimulating factor gene expression in vascular cells and in experimental and human atherosclerosis. Am J Pathol. 1992 Feb;140(2):301–316. [PMC free article] [PubMed] [Google Scholar]
  13. Collins T. Endothelial nuclear factor-kappa B and the initiation of the atherosclerotic lesion. Lab Invest. 1993 May;68(5):499–508. [PubMed] [Google Scholar]
  14. Cybulsky M. I., Gimbrone M. A., Jr Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis. Science. 1991 Feb 15;251(4995):788–791. doi: 10.1126/science.1990440. [DOI] [PubMed] [Google Scholar]
  15. Dartsch P. C., Bauriedel G., Schinko I., Weiss H. D., Höfling B., Betz E. Cell constitution and characteristics of human atherosclerotic plaques selectively removed by percutaneous atherectomy. Atherosclerosis. 1989 Dec;80(2):149–157. doi: 10.1016/0021-9150(89)90023-3. [DOI] [PubMed] [Google Scholar]
  16. Duyao M. P., Buckler A. J., Sonenshein G. E. Interaction of an NF-kappa B-like factor with a site upstream of the c-myc promoter. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4727–4731. doi: 10.1073/pnas.87.12.4727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Edgington T. S., Mackman N., Brand K., Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost. 1991 Jul 12;66(1):67–79. [PubMed] [Google Scholar]
  18. Ferrell M., Fuster V., Gold H. K., Chesebro J. H. A dilemma for the 1990s. Choosing appropriate experimental animal model for the prevention of restenosis. Circulation. 1992 Apr;85(4):1630–1631. doi: 10.1161/01.cir.85.4.1630. [DOI] [PubMed] [Google Scholar]
  19. Fong L. G., Fong T. A., Cooper A. D. Inhibition of lipopolysaccharide-induced interleukin-1 beta mRNA expression in mouse macrophages by oxidized low density lipoprotein. J Lipid Res. 1991 Dec;32(12):1899–1910. [PubMed] [Google Scholar]
  20. Gown A. M., Tsukada T., Ross R. Human atherosclerosis. II. Immunocytochemical analysis of the cellular composition of human atherosclerotic lesions. Am J Pathol. 1986 Oct;125(1):191–207. [PMC free article] [PubMed] [Google Scholar]
  21. Grilli M., Chiu J. J., Lenardo M. J. NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. Int Rev Cytol. 1993;143:1–62. doi: 10.1016/s0074-7696(08)61873-2. [DOI] [PubMed] [Google Scholar]
  22. Hamilton T. A., Ma G. P., Chisolm G. M. Oxidized low density lipoprotein suppresses the expression of tumor necrosis factor-alpha mRNA in stimulated murine peritoneal macrophages. J Immunol. 1990 Mar 15;144(6):2343–2350. [PubMed] [Google Scholar]
  23. Hiscott J., Marois J., Garoufalis J., D'Addario M., Roulston A., Kwan I., Pepin N., Lacoste J., Nguyen H., Bensi G. Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. Mol Cell Biol. 1993 Oct;13(10):6231–6240. doi: 10.1128/mcb.13.10.6231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Iademarco M. F., McQuillan J. J., Rosen G. D., Dean D. C. Characterization of the promoter for vascular cell adhesion molecule-1 (VCAM-1). J Biol Chem. 1992 Aug 15;267(23):16323–16329. [PubMed] [Google Scholar]
  25. Kaltschmidt C., Kaltschmidt B., Henkel T., Stockinger H., Baeuerle P. A. Selective recognition of the activated form of transcription factor NF-kappa B by a monoclonal antibody. Biol Chem Hoppe Seyler. 1995 Jan;376(1):9–16. doi: 10.1515/bchm3.1995.376.1.9. [DOI] [PubMed] [Google Scholar]
  26. Kaltschmidt C., Kaltschmidt B., Lannes-Vieira J., Kreutzberg G. W., Wekerle H., Baeuerle P. A., Gehrmann J. Transcription factor NF-kappa B is activated in microglia during experimental autoimmune encephalomyelitis. J Neuroimmunol. 1994 Nov;55(1):99–106. doi: 10.1016/0165-5728(94)90151-1. [DOI] [PubMed] [Google Scholar]
  27. Kaltschmidt C., Kaltschmidt B., Neumann H., Wekerle H., Baeuerle P. A. Constitutive NF-kappa B activity in neurons. Mol Cell Biol. 1994 Jun;14(6):3981–3992. doi: 10.1128/mcb.14.6.3981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Leenen P. J., de Bruijn M. F., Voerman J. S., Campbell P. A., van Ewijk W. Markers of mouse macrophage development detected by monoclonal antibodies. J Immunol Methods. 1994 Sep 14;174(1-2):5–19. doi: 10.1016/0022-1759(94)90005-1. [DOI] [PubMed] [Google Scholar]
  29. Lenardo M. J., Baltimore D. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell. 1989 Jul 28;58(2):227–229. doi: 10.1016/0092-8674(89)90833-7. [DOI] [PubMed] [Google Scholar]
  30. Levey A. I., Bolam J. P., Rye D. B., Hallanger A. E., Demuth R. M., Mesulam M. M., Wainer B. H. A light and electron microscopic procedure for sequential double antigen localization using diaminobenzidine and benzidine dihydrochloride. J Histochem Cytochem. 1986 Nov;34(11):1449–1457. doi: 10.1177/34.11.2430010. [DOI] [PubMed] [Google Scholar]
  31. Liao F., Andalibi A., Qiao J. H., Allayee H., Fogelman A. M., Lusis A. J. Genetic evidence for a common pathway mediating oxidative stress, inflammatory gene induction, and aortic fatty streak formation in mice. J Clin Invest. 1994 Aug;94(2):877–884. doi: 10.1172/JCI117409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Libby P., Hansson G. K. Involvement of the immune system in human atherogenesis: current knowledge and unanswered questions. Lab Invest. 1991 Jan;64(1):5–15. [PubMed] [Google Scholar]
  33. Mackman N., Brand K., Edgington T. S. Lipopolysaccharide-mediated transcriptional activation of the human tissue factor gene in THP-1 monocytic cells requires both activator protein 1 and nuclear factor kappa B binding sites. J Exp Med. 1991 Dec 1;174(6):1517–1526. doi: 10.1084/jem.174.6.1517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Marui N., Offermann M. K., Swerlick R., Kunsch C., Rosen C. A., Ahmad M., Alexander R. W., Medford R. M. Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. J Clin Invest. 1993 Oct;92(4):1866–1874. doi: 10.1172/JCI116778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Masuda J., Ross R. Atherogenesis during low level hypercholesterolemia in the nonhuman primate. II. Fatty streak conversion to fibrous plaque. Arteriosclerosis. 1990 Mar-Apr;10(2):178–187. doi: 10.1161/01.atv.10.2.178. [DOI] [PubMed] [Google Scholar]
  36. Müller J. M., Ziegler-Heitbrock H. W., Baeuerle P. A. Nuclear factor kappa B, a mediator of lipopolysaccharide effects. Immunobiology. 1993 Apr;187(3-5):233–256. doi: 10.1016/S0171-2985(11)80342-6. [DOI] [PubMed] [Google Scholar]
  37. Nakajima T., Kitajima I., Shin H., Takasaki I., Shigeta K., Abeyama K., Yamashita Y., Tokioka T., Soejima Y., Maruyama I. Involvement of NF-kappa B activation in thrombin-induced human vascular smooth muscle cell proliferation. Biochem Biophys Res Commun. 1994 Oct 28;204(2):950–958. doi: 10.1006/bbrc.1994.2552. [DOI] [PubMed] [Google Scholar]
  38. Neish A. S., Williams A. J., Palmer H. J., Whitley M. Z., Collins T. Functional analysis of the human vascular cell adhesion molecule 1 promoter. J Exp Med. 1992 Dec 1;176(6):1583–1593. doi: 10.1084/jem.176.6.1583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Parhami F., Fang Z. T., Fogelman A. M., Andalibi A., Territo M. C., Berliner J. A. Minimally modified low density lipoprotein-induced inflammatory responses in endothelial cells are mediated by cyclic adenosine monophosphate. J Clin Invest. 1993 Jul;92(1):471–478. doi: 10.1172/JCI116590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Poston R. N., Haskard D. O., Coucher J. R., Gall N. P., Johnson-Tidey R. R. Expression of intercellular adhesion molecule-1 in atherosclerotic plaques. Am J Pathol. 1992 Mar;140(3):665–673. [PMC free article] [PubMed] [Google Scholar]
  41. Qwarnström E. E., Ostberg C. O., Turk G. L., Richardson C. A., Bomsztyk K. Fibronectin attachment activates the NF-kappa B p50/p65 heterodimer in fibroblasts and smooth muscle cells. J Biol Chem. 1994 Dec 9;269(49):30765–30768. [PubMed] [Google Scholar]
  42. Rajavashisth T. B., Andalibi A., Territo M. C., Berliner J. A., Navab M., Fogelman A. M., Lusis A. J. Induction of endothelial cell expression of granulocyte and macrophage colony-stimulating factors by modified low-density lipoproteins. Nature. 1990 Mar 15;344(6263):254–257. doi: 10.1038/344254a0. [DOI] [PubMed] [Google Scholar]
  43. Rosenfeld M. E., Ylä-Herttuala S., Lipton B. A., Ord V. A., Witztum J. L., Steinberg D. Macrophage colony-stimulating factor mRNA and protein in atherosclerotic lesions of rabbits and humans. Am J Pathol. 1992 Feb;140(2):291–300. [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Terkeltaub R., Banka C. L., Solan J., Santoro D., Brand K., Curtiss L. K. Oxidized LDL induces monocytic cell expression of interleukin-8, a chemokine with T-lymphocyte chemotactic activity. Arterioscler Thromb. 1994 Jan;14(1):47–53. doi: 10.1161/01.atv.14.1.47. [DOI] [PubMed] [Google Scholar]
  46. Weber C., Erl W., Pietsch A., Ströbel M., Ziegler-Heitbrock H. W., Weber P. C. Antioxidants inhibit monocyte adhesion by suppressing nuclear factor-kappa B mobilization and induction of vascular cell adhesion molecule-1 in endothelial cells stimulated to generate radicals. Arterioscler Thromb. 1994 Oct;14(10):1665–1673. doi: 10.1161/01.atv.14.10.1665. [DOI] [PubMed] [Google Scholar]
  47. Wilcox J. N., Smith K. M., Schwartz S. M., Gordon D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2839–2843. doi: 10.1073/pnas.86.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Witztum J. L., Steinberg D. Role of oxidized low density lipoprotein in atherogenesis. J Clin Invest. 1991 Dec;88(6):1785–1792. doi: 10.1172/JCI115499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wu H., Moulton K., Horvai A., Parik S., Glass C. K. Combinatorial interactions between AP-1 and ets domain proteins contribute to the developmental regulation of the macrophage scavenger receptor gene. Mol Cell Biol. 1994 Mar;14(3):2129–2139. doi: 10.1128/mcb.14.3.2129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Ylä-Herttuala S., Lipton B. A., Rosenfeld M. E., Särkioja T., Yoshimura T., Leonard E. J., Witztum J. L., Steinberg D. Expression of monocyte chemoattractant protein 1 in macrophage-rich areas of human and rabbit atherosclerotic lesions. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5252–5256. doi: 10.1073/pnas.88.12.5252. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES