Summary
We have developed a solid-phase enzyme immunoassay for anti-oxidized low-density lipoprotein antibodies. Most sera showed some degree of non-specific binding to plates coated with oxidized low-density lipoprotein and the autoantibodies to oxidized low-density lipoprotein often appeared to have a relatively low affinity. To differentiate between specific and non-specific binding each sample was tested untreated and after absorption with oxidized low-density lipoprotein. The optical densities obtained with dilutions of the absorbed sample were considered to reflect non-specific binding and were subtracted from values obtained with identical dilutions of the unabsorbed sample, to yield corrected values from which the concentrations of anti-oxidized low-density lipoprotein antibody were calculated. Similar absorptions with native low-density lipoprotein and oxidized human serum albumin failed to induce a significant reduction in binding to immobilized oxidized low-density lipoprotein proving that the antibodies measured by this assay are primarily specific for oxidized low-density lipoprotein. We studied sera from two groups of individuals: (1) 33 subjects submitted to coronary angiography and split into two subgroups depending on the degree of coronary stenosis and (2) 64 healthy individuals also split into two subgroups according to lipid levels. Anti-oxidized low-density lipoprotein antibodies were detected both in patients and healthy individuals. Higher levels were detected in patients with moderate coronary disease and hyperlipemic healthy individuals, but the differences between patients and healthy volunteers or between their respective subgroups did not reach statistical significance. Our results suggest that autoantibodies to oxidized low-density lipoprotein are relatively frequent in both symptomatic and asymptomatic individuals. The investigation of their potential role as a risk factor will require mass screening and long-term follow-up.
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Basu SK, Goldstein JL, Anderson RGW, Brown MS, Degradation of cationized LDL and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts. Proc Natl Acad Sci USA 73:178, 1976
Beaumont JL, Autoimmune hyperlipidemia. An atherogenic metabolic disease of immune origin. Rev Eur Stud Clin Bio 15:1037, 1970
Beaumont JL, Beaumont V, Immunological aspects of atherosclerosis. Atherosclerosis Rev 3:133, 1978
Bérard M, Antonucci M, Beaumont JL, Cytotoxic effect of serum on fibrablasts in one case of normolipidemic plane xantoma and myeloma IgG 1. Atherosclerosis 62:111, 1986
Berliner JA, Territo MC, Sevanian A, Ramin S, Kim JA, Bamshad B, Esterson M, Fogelman A, Minimally modified low density lipoprotein stimulates monocyte-endothelial interactions. J Clin Invest 85:1260, 1990
Cazzolato G, Avogaro P, Bittolo-Bon G, Characterization of a more electronegative charged LDL subfraction by ion exchange HPLC. Free Radic Biol Med 11:247, 1991
Dachet C, Bandet MF, Beaumont JL, Cholesterol synthesis by human fibroblasts in the presence of LDL and anti-LDL IgA. Biomedicine 31:80, 1979
Feingold KR, Castro GR, Ishikawa Y, Fielding PE, Fielding CJ, Cutaneous xantoma in association with paraproteinemia in the absence of hyperlipidemia. J Clin Invest 83:796, 1989
Füst G, Szondy E, Szekely J, Nanai I, Gerö S, Studies on the occurrence of circulating immune complexes in vascular disease. Arteriosclerosis 29:181, 1978
Gero S, Szondy E, Mezey Z, Székely J, Immune response against lipoproteins in coronary patients. In: Latent dyslipoproteinemias and arteriosclerosis. DeGennes JL (ed) Raven, New York, pp 73–77, 1984
Gissinger C, Virella GT, Lopes-Virella MF, Eryhtrocyte-bound low-density lipoprotein immune complexes lead to cholesteryl ester accumulation in human monocyte-derived macrophages. Clin Immunol Immunopathol 59:37, 1991
Griffith RL, Virella GT, Stevensen HC, Lopes-Virella MF, Low density lipoprotein metabolism by human macrophages activated with low density lipoprotein immune complexes. J Exp Med 168:1041, 1988
Kim RS, LaBella FS, Comparison of analytical methods for monitoring autoxidation profiles of authentic lipids. J Lipid Res 28:1110, 1987
Klimov AN, Denisenko AD, Popov AV, Nagornev VA, Pleskov VM, Vinogradov AG, Denisenko TV, Ya Magracheva E, Kheifes GM, Kuznetzov AS, Lipoprotein-antibody immune complexes: their catabolism and role in foam cell formation. Atherosclerosis 58:1, 1985
Lopes-Virella MF, Sherer GK, Lees AM, Wohltmann H, Mayfield R, Sagel J, Leroy C, Colwell JA, Surface binding, internalization, and degradation by cultured human fibroblasts of LDL isolated from type 1 (insulin-dependent) diabetic patients: changes with metabolic control. Diabetologia 22:430, 1982
Lorenzelli-Edouard L, Beaumont FM, Beaumont JL, Antilipoprotein autoimmune hyperlipidemia. Biomedicine 33:160, 1980
Morel DW, Hessler JR, Chisolm GM, Low density lipoprotein cytotoxicity induced by free-radical peroidation of lipid. J Lipid Res 24:1070, 1983
Orekhov AN, Tertov VV, Kabakov AE, Yu I, Pokrovsky SN, Smirnov VN, Autoantibodies against modified lipoprotein. Nonlipid factor of blood plasma that stimulated foam cell formation. Arteriosclerosis Thromb 11:316, 1991
Palinski W, Rosenfeld ME, Ylä-Herttuala S, Gurtner GC, Socher SS, Butler SW, Pathasarathy S, Carew TE, Steinberg D, Witztum JL, Low density lipoprotein undergoes oxidative modificationin vivo. Proc Natl Acad Sci USA 86:1372, 1989
Parums DV, Mitchinson MJ, Serum antibodies to oxidized LDL and ceroid in chronic periaortitis. Am J Pathol 151:57A, 1987
Pestel J, Joseph M, Dessaint JP, Capron A, Macrophage triggering by aggregated immunoglobulin. I. Delayed effect of IgG aggregates or immune complexes. J Immunol 126:1887, 1981
Riesen W, Noseda G, Antibodies against lipoproteins in man. Occurrence and biological significance. Klin Wochenschr 53:353, 1975
Riesen W, Noseda G, Butler R, Anti-β-lipoprotein activity of human monoclonal immunoglobulins. Vox Sang 22:420, 1972
Salmon S, Maziere C, Theron L, Beucer I, Ayault-Jarrier M, Goldstein S, Polonovski J, Immunological detection of lowdensity lipoproteins modified by malondialdehydein vitro orin vivo. Biochim. Biophys Acta 920:215, 1987
Salonen JT, Ÿla-Herttuala S, Yamamoto R, Butler S, Korpela H, Salonen R, Nyyssönen K, Palinsky W, Witzum J, Autoantibody against oxidised LDL and progression of carotid atherosclerosis. Lancet 339:883, 1992
Steinbrecher UP, Fisher M, Witztum JL, Curtiss LK, Immunogenicity of homologous low density lipoprotein after methylation, ethylation or carbamylation: generation of antibodies specific for derivatized lysine. J Lipid Res 25:1109, 1984
Steinbrecher UP, Parthasaranthy S, Leake DS, Witztum JL, Steinberg D, Modification of low density lipoprotein by endothelial cells involves lipid peroxidation and degradation of low density lipoprotein phospholipids, Proc Natl Acad Sci USA 81:3883, 1984
Szondy E, Horvath M, Mezey Z, Szekely J, Lengyel E, Gero S, Free and complexed anti-lipoprotein antibodies in vascular disease. Atherosclerosis 49:69, 1983
Szondy E, Lengyel E, Mezey Z, Fust G, Gero S, Occurrence of anti-low density lipoprotein antibodies and circulating immune complexes in aged subjects. Mech Aging Dev 29:117, 1985
Witztum JL, The role of oxidized LDL in atherosclerosis. In: Hypercholesterolemia, hypocholesterolemia, hypertriglyceridemia. Malmendier CL (ed) Plenum, New York, p 353, 1990
Witzum JL, Steinbrecher UP, Kesaniemi YA, Fisher M, Autoantibodies to glucosylated proteins in the plasma of patients with diabetes mellitus. Proc Natl Acad Sci USA 81:3204, 1984
Ylä-Herttuala S, Palinski W, Rosenfeld ME, Steinberg D, Witzum JL, Lipoproteins in normal and atherosclerotic aorta. Eur Heart J 11 [Suppl E]:88, 1990
Ylä-Herttuala S, Rosenfeld ME, Parthasarathy S, Glass CK, Sigal E, Witztum JL, Steinberg D, Co-localization of 15-lipoxygenase mRNA and protein with epitopes of oxidized low density lipoprotein in macrophage-rich areas of atherosclerotic lesions. Proc Natl Acad Sci USA 87:6959, 1990
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Virella, G., Virella, I., Leman, R.B. et al. Anti-oxidized low-density lipoprotein antibodies in patients with coronary heart disease and normal healthy volunteers. Int J Clin Lab Res 23, 95–101 (1993). https://doi.org/10.1007/BF02592290
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DOI: https://doi.org/10.1007/BF02592290