Arteriosclerosis, Vol 10, 325-335, Copyright © 1990 by American Heart Association

ARTICLES

Antisera and monoclonal antibodies specific for epitopes generated during oxidative modification of low density lipoprotein

W Palinski, S Yla-Herttuala, ME Rosenfeld, SW Butler, SA Socher, S Parthasarathy, LK Curtiss and JL Witztum
Department of Medicine, University of California-San Diego, La Jolla 92093.

Increasing evidence indicates that low density lipoprotein (LDL) has to be modified to induce foam cell formation. One such modification, oxidation of LDL, generates a number of highly reactive short chain- length aldehydic fragments of oxidized fatty acids capable of conjugating with lysine residues of apoprotein B. By immunizing animals with homologous malondialdehyde-modified LDL (MDA-LDL), 4- hydroxynonenal-LDL (4-HNE-LDL), and Cu+(+)-oxidized LDL, we developed polyvalent and monoclonal antibodies against three epitopes found in oxidatively modified LDL. The present article characterizes an antiserum and monoclonal antibody (MAL-2 and MDA2, respectively) specific for MDA-lysine, and an antiserum and monoclonal antibody (HNE- 6 and NA59, respectively) specific for 4-HNE-lysine. In addition, a monoclonal antibody (OLF4-3C10) was developed against an as yet undefined epitope generated during Cu++ oxidation of LDL. With these antibodies, we demonstrated that MDA-lysine and 4-HNE-lysine adducts develop on apo-lipoprotein B during copper-induced oxidation of LDL in vitro. The application of these antibodies for immunocytochemical demonstration of oxidized lipoproteins in atherosclerotic lesions of progressive severity is described in the companion article. These antibodies should prove useful in studying the role of oxidatively modified lipoproteins as well as other oxidatively modified proteins in atherogenesis.

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				S. Du Yan, Y. Shi, A. Zhu, J. Fu, H. Zhu, Y. Zhu, L. Gibson, E. Stern, K. Collison, F. Al-Mohanna, et al. Role of ERAB/L-3-Hydroxyacyl-coenzyme A Dehydrogenase Type II Activity in Abeta -induced Cytotoxicity J. Biol. Chem., January 22, 1999; 274(4): 2145 - 2156. [Abstract] [Full Text] [PDF]

				R. Wu, U. de Faire, C. Lemne, J. L. Witztum, and J. Frostegard Autoantibodies to OxLDL Are Decreased in Individuals With Borderline Hypertension Hypertension, January 1, 1999; 33(1): 53 - 59. [Abstract] [Full Text] [PDF]

				S. Freigang, S. Horkko, E. Miller, J. L. Witztum, and W. Palinski Immunization of LDL Receptor–Deficient Mice With Homologous Malondialdehyde-Modified and Native LDL Reduces Progression of Atherosclerosis by Mechanisms Other Than Induction of High Titers of Antibodies to Oxidative Neoepitopes Arterioscler Thromb Vasc Biol, December 1, 1998; 18(12): 1972 - 1982. [Abstract] [Full Text] [PDF]

				J. Hulthe, J. Wikstrand, A. Lidell, I. Wendelhag, G. K. Hansson, and O. Wiklund Antibody Titers Against Oxidized LDL Are Not Elevated in Patients With Familial Hypercholesterolemia Arterioscler Thromb Vasc Biol, August 1, 1998; 18(8): 1203 - 1211. [Abstract] [Full Text] [PDF]

				M. Ricote, J. Huang, L. Fajas, A. Li, J. Welch, J. Najib, J. L. Witztum, J. Auwerx, W. Palinski, and C. K. Glass Expression of the peroxisome proliferator-activated receptor gamma  (PPARgamma ) in human atherosclerosis and regulation in macrophages by colony stimulating factors and oxidized low density lipoprotein PNAS, June 23, 1998; 95(13): 7614 - 7619. [Abstract] [Full Text] [PDF]

				F. Calara, P. Dimayuga, A. Niemann, J. Thyberg, U. Diczfalusy, J. L. Witztum, W. Palinski, P. K. Shah, B. Cercek, J. Nilsson, et al. An Animal Model to Study Local Oxidation of LDL and Its Biological Effects in the Arterial Wall Arterioscler Thromb Vasc Biol, June 1, 1998; 18(6): 884 - 893. [Abstract] [Full Text] [PDF]

				M. Torzewski, M. Klouche, J. Hock, M. Meßner, B. Dorweiler, J. Torzewski, H. E. Gabbert, and S. Bhakdi Immunohistochemical Demonstration of Enzymatically Modified Human LDL and Its Colocalization With the Terminal Complement Complex in the Early Atherosclerotic Lesion Arterioscler Thromb Vasc Biol, March 1, 1998; 18(3): 369 - 378. [Abstract] [Full Text] [PDF]

				J. S. Luoma, P. Stralin, S. L. Marklund, T. P. Hiltunen, T. Sarkioja, and S. Yla-Herttuala Expression of Extracellular SOD and iNOS in Macrophages and Smooth Muscle Cells in Human and Rabbit Atherosclerotic Lesions : Colocalization With Epitopes Characteristic of Oxidized LDL and Peroxynitrite-Modified Proteins Arterioscler Thromb Vasc Biol, February 1, 1998; 18(2): 157 - 167. [Abstract] [Full Text] [PDF]

				P. Reaven, S. Merat, F. Casanada, M. Sutphin, and W. Palinski Effect of Streptozotocin-Induced Hyperglycemia on Lipid Profiles, Formation of Advanced Glycation Endproducts in Lesions, and Extent of Atherosclerosis in LDL Receptor-Deficient Mice Arterioscler Thromb Vasc Biol, October 1, 1997; 17(10): 2250 - 2256. [Abstract] [Full Text]

				S. Horkko, E. Miller, D. W. Branch, W. Palinski, and J. L. Witztum The epitopes for some antiphospholipid antibodies are adducts of oxidized phospholipid and beta 2 glycoprotein 1 (and other proteins) PNAS, September 16, 1997; 94(19): 10356 - 10361. [Abstract] [Full Text] [PDF]

				C. Iribarren, A. R. Folsom, D. R. Jacobs Jr, M. D. Gross, J. D. Belcher, and J. H. Eckfeldt Association of Serum Vitamin Levels, LDL Susceptibility to Oxidation, and Autoantibodies Against MDA-LDL With Carotid Atherosclerosis: A Case-Control Study Arterioscler Thromb Vasc Biol, June 1, 1997; 17(6): 1171 - 1177. [Abstract] [Full Text]

				M. Seccia, E. Albano, E. Maggi, and G. Bellomo Circulating Autoantibodies Recognizing Peroxidase-Oxidized Low Density Lipoprotein: Evidence for New Antigenic Epitopes Formed In Vivo Independently From Lipid Peroxidation Arterioscler Thromb Vasc Biol, January 1, 1997; 17(1): 134 - 140. [Abstract] [Full Text]

				M. I.J. Uusitupa, L. Niskanen, J. Luoma, P. Vilja, M. Mercuri, R. Rauramaa, and S. Yla-Herttuala Autoantibodies Against Oxidized LDL Do Not Predict Atherosclerotic Vascular Disease in Non–Insulin-Dependent Diabetes Mellitus Arterioscler Thromb Vasc Biol, October 1, 1996; 16(10): 1236 - 1242. [Abstract] [Full Text]

				K. D. O'Brien, T. O. McDonald, A. Chait, M. D. Allen, and C. E. Alpers Neovascular Expression of E-Selectin, Intercellular Adhesion Molecule-1, and Vascular Cell Adhesion Molecule-1 in Human Atherosclerosis and Their Relation to Intimal Leukocyte Content Circulation, February 15, 1996; 93(4): 672 - 682. [Abstract] [Full Text]

				M. Mironova, G. Virella, and M. F. Lopes-Virella Isolation and Characterization of Human Antioxidized LDL Autoantibodies Arterioscler Thromb Vasc Biol, February 1, 1996; 16(2): 222 - 229. [Abstract] [Full Text]

				W. Palinski, R. K. Tangirala, E. Miller, S. G. Young, and J. L. Witztum Increased Autoantibody Titers Against Epitopes of Oxidized LDL in LDL Receptor–Deficient Mice With Increased Atherosclerosis Arterioscler Thromb Vasc Biol, October 1, 1995; 15(10): 1569 - 1576. [Abstract] [Full Text]

				R. K. Tangirala, F. Casanada, E. Miller, J. L. Witztum, D. Steinberg, and W. Palinski Effect of the Antioxidant N,N'-Diphenyl 1,4-Phenylenediamine (DPPD) on Atherosclerosis in ApoE-Deficient Mice Arterioscler Thromb Vasc Biol, October 1, 1995; 15(10): 1625 - 1630. [Abstract] [Full Text]

				P. Lesnik, C. Dentan, A. Vonica, M. Moreau, and M. J. Chapman Tissue Factor Pathway Inhibitor Activity Associated With LDL Is Inactivated by Cell- and Copper-Mediated Oxidation Arterioscler Thromb Vasc Biol, August 1, 1995; 15(8): 1121 - 1130. [Abstract] [Full Text]

				P. A. Kiener, B. M. Rankin, P. M. Davis, S. A. Yocum, G. A. Warr, and R. I. Grove Immune Complexes of LDL Induce Atherogenic Responses in Human Monocytic Cells Arterioscler Thromb Vasc Biol, July 1, 1995; 15(7): 990 - 999. [Abstract] [Full Text]

				W. Palinski, T. Koschinsky, S. W. Butler, E. Miller, H. Vlassara, A. Cerami, and J. L. Witztum Immunological Evidence for the Presence of Advanced Glycosylation End Products in Atherosclerotic Lesions of Euglycemic Rabbits Arterioscler Thromb Vasc Biol, May 1, 1995; 15(5): 571 - 582. [Abstract] [Full Text]

				A. Hammer, G. Kager, G. Dohr, H. Rabl, I. Ghassempur, and G. Jurgens Generation, Characterization, and Histochemical Application of Monoclonal Antibodies Selectively Recognizing Oxidatively Modified ApoB-Containing Serum Lipoproteins Arterioscler Thromb Vasc Biol, May 1, 1995; 15(5): 704 - 713. [Abstract] [Full Text]

				C. Bergmark, R. Wu, U. d. Faire, A. K. Lefvert, and J. Swedenborg Patients With Early-Onset Peripheral Vascular Disease Have Increased Levels of Autoantibodies Against Oxidized LDL Arterioscler Thromb Vasc Biol, April 1, 1995; 15(4): 441 - 445. [Abstract] [Full Text]

				A. Philis-Tsimikas, S. Parthasarathy, S. Picard, W. Palinski, and J. L. Witztum Aminoguanidine Has Both Pro-oxidant and Antioxidant Activity Toward LDL Arterioscler Thromb Vasc Biol, March 1, 1995; 15(3): 367 - 376. [Abstract] [Full Text]

				H. A. Kleinveld, H. L.M. Hak-Lemmers, M. P.C. Hectors, N. J. de Fouw, P. N.M. Demacker, and A. F.H. Stalenhoef Vitamin E and Fatty Acid Intervention Does Not Attenuate the Progression of Atherosclerosis in Watanabe Heritable Hyperlipidemic Rabbits Arterioscler Thromb Vasc Biol, February 1, 1995; 15(2): 290 - 297. [Abstract] [Full Text]

				M. Puurunen, M. Manttari, V. Manninen, L. Tenkanen, G. Alfthan, C. Ehnholm, O. Vaarala, K. Aho, and T. Palosuo Antibody Against Oxidized Low-Density Lipoprotein Predicting Myocardial Infarction Arch Intern Med, November 28, 1994; 154(22): 2605 - 2609. [Abstract] [PDF]