Inhibition of Nuclear Factor-B–Mediated Adhesion Molecule Expression in Human Endothelial Cells

From the Cardiology Section (J.M.L., J.S.C., W.L.A.-L., A.J.B.) and Nephrology Section (T.H.), Department of Medicine, and the Molecular and Cellular Biology Graduate Program (J.M.L., J.S.C.), Tulane University Medical School, and the VA Medical Center (T.H.), New Orleans, La.

Correspondence to Jean M. Lockyer, PhD, Cardiology Section SL48, Tulane University Medical School, 1430 Tulane Ave, New Orleans, LA 70112-2699. E-mail Lockyer.Jean{at}tmc.tulane.edu

Abstract—The transcriptional regulatory protein nuclear factor-B (NF-B) participates in the control of gene expression of many modulators of the inflammatory and immune responses, including the adhesion molecules E-selectin and intercellular adhesion molecule-1 (ICAM-1). NF-B is found in the cytoplasm complexed with its inhibitory protein IB. On activation, IB is phosphorylated and degraded, thereby freeing NF-B for translocation to the nucleus. We have generated populations of endothelial cells expressing wild-type and a proteolysis-resistant mutation of IB that is lacking the 36 N-terminal amino acids (IBN) in order to examine the effects of expression of the mutated IB on tumor necrosis factor- (TNF-)–induced E-selectin and ICAM-1 expression. Wild-type and IBN were introduced into primary endothelial cells using retrovirus infection followed by selection with G418. The IBN protein remained at untreated control levels in endothelial cells treated with TNF- and also remained complexed with the NF-B family member p65. Furthermore, TNF-–induced NF-B DNA binding activity was inhibited in the population of endothelial cells expressing IBN. That population of cells was also refractory to upregulation of E-selectin and ICAM-1 after treatment with TNF-. The use of a truncated IB protein to prevent NF-B–mediated gene expression provides a novel and specific approach for investigating the role of NF-B in processes associated with adhesion molecule expression during inflammation.

Key Words: nuclear factor-B • vascular endothelium • adhesion molecule • inflammation • gene expression

This article has been cited by other articles:

				M. Rajesh, P. Mukhopadhyay, S. Batkai, G. Hasko, L. Liaudet, J. W. Huffman, A. Csiszar, Z. Ungvari, K. Mackie, S. Chatterjee, et al. CB2-receptor stimulation attenuates TNF-{alpha}-induced human endothelial cell activation, transendothelial migration of monocytes, and monocyte-endothelial adhesion Am J Physiol Heart Circ Physiol, October 1, 2007; 293(4): H2210 - H2218. [Abstract] [Full Text] [PDF]

				R. Williams, X. Zou, and G. W. Hoyle Tachykinin-1 receptor stimulates proinflammatory gene expression in lung epithelial cells through activation of NF-{kappa}B via a Gq-dependent pathway Am J Physiol Lung Cell Mol Physiol, February 1, 2007; 292(2): L430 - L437. [Abstract] [Full Text] [PDF]

				R. Lv, Z.-Q. Zhou, H.-W. Wu, Y. Jin, W. Zhou, and J.-G. Xu Hydroxyethyl starch exhibits antiinflammatory effects in the intestines of endotoxemic rats. Anesth. Analg., July 1, 2006; 103(1): 149 - 155. [Abstract] [Full Text] [PDF]

				G. Rajashekhar, A. Loganath, A. C. Roy, S. S. Chong, and Y. C. Wong Hypoxia Up-regulated Angiogenin and Down-regulated Vascular Cell Adhesion Molecule-i Expression and Secretion in Human Placental Trophoblasts Reproductive Sciences, July 1, 2005; 12(5): 310 - 319. [Abstract] [PDF]

				R. B. Sharma, J. D. Alegria, M. V. Talor, N. R. Rose, P. Caturegli, and C. L. Burek Iodine and IFN-{gamma} Synergistically Enhance Intercellular Adhesion Molecule 1 Expression on NOD.H2h4 Mouse Thyrocytes J. Immunol., June 15, 2005; 174(12): 7740 - 7745. [Abstract] [Full Text] [PDF]

				R. Lv, W. Zhou, C. Chu, and J. Xu Mechanism of the Effect of Hydroxyethyl Starch on Reducing Pulmonary Capillary Permeability in a Rat Model of Sepsis Ann. Clin. Lab. Sci., April 1, 2005; 35(2): 174 - 183. [Abstract] [Full Text] [PDF]

				K. V. RAMANA, A. BHATNAGAR, and S. K. SRIVASTAVA Inhibition of aldose reductase attenuates TNF-{alpha}-induced expression of adhesion molecules in endothelial cells FASEB J, August 1, 2004; 18(11): 1209 - 1218. [Abstract] [Full Text] [PDF]

				J. H. Von der Thusen, J. Kuiper, T. J. C. Van Berkel, and E. A. L. Biessen Interleukins in Atherosclerosis: Molecular Pathways and Therapeutic Potential Pharmacol. Rev., March 1, 2003; 55(1): 133 - 166. [Abstract] [Full Text] [PDF]

				H. Fan, B. Sun, Q. Gu, A. Lafond-Walker, S. Cao, and L. C. Becker Oxygen radicals trigger activation of NF-kappa B and AP-1 and upregulation of ICAM-1 in reperfused canine heart Am J Physiol Heart Circ Physiol, May 1, 2002; 282(5): H1778 - H1786. [Abstract] [Full Text] [PDF]

				J. Kaur, R. C. Woodman, L. Ostrovsky, and P. Kubes Selective recruitment of neutrophils and lymphocytes by thrombin: a role for NF-{kappa}B Am J Physiol Heart Circ Physiol, August 1, 2001; 281(2): H784 - H795. [Abstract] [Full Text] [PDF]

				L. J. Thompson, J. Jiang, N. Madamanchi, M. S. Runge, and C. Patterson PTP-{epsilon}, a tyrosine phosphatase expressed in endothelium, negatively regulates endothelial cell proliferation Am J Physiol Heart Circ Physiol, July 1, 2001; 281(1): H396 - H403. [Abstract] [Full Text] [PDF]

				D. N. Muller, E. M. A. Mervaala, F. Schmidt, J.-K. Park, R. Dechend, E. Genersch, V. Breu, B.-M. Loffler, D. Ganten, W. Schneider, et al. Effect of Bosentan on NF-{kappa}B, Inflammation, and Tissue Factor in Angiotensin II-Induced End-Organ Damage Hypertension, August 1, 2000; 36(2): 282 - 290. [Abstract] [Full Text] [PDF]

				H. C. A. DREXLER, W. RISAU, and M. A. KONERDING Inhibition of proteasome function induces programmed cell death in proliferating endothelial cells FASEB J, January 1, 2000; 14(1): 65 - 77. [Abstract] [Full Text]

				H. Huang, T. M. Calderon, J. W. Berman, V. L. Braunstein, L. M. Weiss, M. Wittner, and H. B. Tanowitz Infection of Endothelial Cells with Trypanosoma cruzi Activates NF-kappa B and Induces Vascular Adhesion Molecule Expression Infect. Immun., October 1, 1999; 67(10): 5434 - 5440. [Abstract] [Full Text] [PDF]

				B. Han and C. D. Logsdon Cholecystokinin induction of mob-1 chemokine expression in pancreatic acinar cells requires NF-kappa B activation Am J Physiol Cell Physiol, July 1, 1999; 277(1): C74 - C82. [Abstract] [Full Text] [PDF]

				K. S.C. Weber, G. Draude, W. Erl, R. de Martin, and C. Weber Monocyte Arrest and Transmigration on Inflamed Endothelium in Shear Flow Is Inhibited by Adenovirus-Mediated Gene Transfer of Ikappa B-alpha Blood, June 1, 1999; 93(11): 3685 - 3693. [Abstract] [Full Text] [PDF]

				S. Kokura, R. E. Wolf, T. Yoshikawa, D. N. Granger, and T. Y. Aw Molecular Mechanisms of Neutrophil-Endothelial Cell Adhesion Induced by Redox Imbalance Circ. Res., March 19, 1999; 84(5): 516 - 524. [Abstract] [Full Text] [PDF]