Evaluation of Autoantibodies Against Oxidized LDL (oLAB) and Blood Antioxidant Status in Professional Soccer Players

Abstract

Low-density lipoproteins (LDL) are very sensitive to oxidative processes initiated by oxygen free radicals, known to be produced in large quantities during intense physical exercise. Oxidatively modified lipoprotein particles (oxLDL) are strongly atherogenic and immunogenic, as a consequence specific autoantibodies (oLAB) against oxLDL are produced by the immune system. This study was designed to evaluate the oLAB titres in professional soccer players and to find out whether the immune response to oxidative modification of LDL correlates with the antioxidant status of individual players. Eleven players volunteered to participate in an incremental treadmill running exercise to volitional fatigue twice (in October and January) during the competitive season. Venous blood samples were withdrawn before and 3 min after the cessation of the test. Serum levels of oLAB were measured by ELISA (Biomedica). Blood samples were analyzed for glutathione peroxidase, reduced glutathione, superoxide dismutase, catalase and glutathione reductase. The activity of creatine kinase (CK) and concentrations of malondialdehyde (MDA), vitamin E and retinol were determined in plasma. From 11 subjects only in 4 players, in both graded running tests, the oLAB titres were low (< 200 mU · ml^-1). The remaining athletes presented elevated oLAB (800 - 1400 mU · ml^-1). Significantly lower activities of catalase and glutathione reductase and lower concentration of α-tocopherol were recorded in the 2nd trial. When the data were arranged according to the oLAB titres no significant between-group differences were found in either pre- and post-test activities of antioxidant enzymes or in concentrations of antioxidants. However, significantly higher CK activities and a tendency towards more elevated plasma MDA concentrations were observed in subjects with higher oLAB levels. It seems justified to presume that high titres of antibodies against oxLDL, as evidenced in most of the players, could be accounted for by their higher in vivo susceptibility of LDL to structural modification under conditions of intensive training-induced oxidative stress, despite their apparently normal antioxidant status.

Autoantibodies against oxidized LDL - antioxidant enzymes - vitamin E

References

• 1 Aebi H. Catalase. Bergmeyer HO Methods of Enzymatic Analysis. New York; Academic Press 1974: 673-683
• 2 Bender D A, Bender A E. Nutrition: A Reference Book. Oxford; Oxford University Press 1997: 228-244
• 3 Beutler E, Duron O, Mikus Kelly B. Improved method for the determination of blood glutathione.  J Lab Clin Med. 1963;  61 882-888
• 4 Brigelius-Flohé R, Traber M G. Vitamin E: function and metabolism.  FASEB J. 1999;  13 1145-1155
• 5 Brites F D, Evelson P A, Christiansen M G, Nicol M F, Basilico M J, Wikinski R W, Llesuy S F. Soccer players under regular training show oxidative stress but improved plasma antioxidant status.  Clin Sci. 1999;  96 381-385
• 6 Buege J A, Aust S D. Microsomal lipid peroxidation.  Meth Enzymol. 1978;  52 302-310
• 7 Clarkson P M, Sayers S P. .  Etiology of exercise-induced muscle damage Can J Appl Physiol. 1999;  24 234-248
• 8 Clarkson P M, Thompson H S. Antioxidants: what role do they play in physical activity and health?.  Am J Clin Nutr. 2000;  72 637-646
• 9 Evelson P, Gambino G, Travacio M, Jaita G, Verona J, Maroncelli C, Wikinski R, Liesuy S, Brites F. Higher antioxidant defences in plasma and low density lipoproteins from rugby players.  Eur J Clin Invest. 2002;  32 818-825
• 10 Fossati P, Prencipe L, Berti G. Use of 3, 5-dichloro-2-hydroxybenzenosulfonic acid/4-aminophenazone chromogenic system in direct enzymic assay of uric acid in serum and urine.  Clin Chem. 1980;  26 227-231
• 11 Fukumoto M, Shoji T, Emoto M, Kawagishi T, Okuno Y, Nishizawa Y. Antibodies against oxidized LDL and carotid artery intima-media thickness in a healthy population.  Arterioscler Tromb Vasc Biol. 2000;  20 703-707
• 12 Glatzle G, Korner W F, Christeller S, Wiss O. Method for the detection of a biochemical riboflavin deficiency. Stimulation of NADPH₂-dependent glutathione reductase from human erythrocytes by FAD in vitro. Investigations on the vitamin B₂ status in healthy people and geriatric patients.  Intern J Vit Res. 1970;  40 166-183
• 13 Halliwell B, Clement M V, Long L H. Hydrogen peroxide in the human body.  FEBS Lett. 2000;  486 10-13
• 14 Heitzer T, Herttuala S Y, Wild E, Luoma J, Drexler H. Effect of vitamin E on endothelial vasodilator function in patients with hypercholerolemia, chronic smoking or both.  J Am Coll Cardiol. 1999;  33 499-505
• 15 Hellsten Y, Svensson M, Sjödin B, Smith S, Christensen A, Richter E A, Bangsbo J. Allantoin formation and urate and glutathione exchange in human muscle during submaximal exercise.  Free Radic Biol Med. 2001;  31 1313-1322
• 16 Houston M, Estevez A, Chumley P, Aslan M, Marklund S, Parks D A, Freeman B A. Binding of xanthine oxidase to vascular endothelium.  J Biol Chem. 1999;  274 4985-4994
• 17 Inoue T, Uchida T, Kamishirado H, Takayanagi K, Hayashi T, Morooka S. Clinical significance of antibody against oxidized low density lipoprotein in patients with atherosclerotic coronary artery disease.  J Am Coll Cardiol. 2001;  37 775-559
• 18 Iribarren C, Folsom A R, Jacobs D R, Gross M D, Belcher J D, Eckfeldt J H. Association of serum vitamin levels, LDL susceptibility to oxidation, and autoantibodies against MDA-LDL with carotid atherosclerosis. A case-control study.  Arterioscl Tromb Vasc Biol. 1997;  17 1171-1177
• 19 Jialal I, Fuller C J, Huet B A. The effect of α-tocopherol supplementation on LDL oxidation.  Arterioscler Thromb Vasc Biol. 1995;  15 190-198
• 20 Kaul N, Devaraj S, Jialal I. α-Tocopherol and atherosclerosis.  Exp Biol Med. 2001;  226 5-12
• 21 Lacy F, Gough D A, Schmid-Schonbein G W. Role of xanthine oxidase in hydrogen peroxide production.  Free Radic Biol Med. 1998;  25 720-727
• 22 Liu M L, Bergholm R, Mäkimattila S, Lahdenperä S, Valkonen M, Hilden Y, Yki-Järvinen H, Taskinen M R. A marathon run increases the susceptibility of LDL to oxidation in vitro and modifies plasma antioxidants.  Am J Physiol. 1999;  276 1083-1091
• 23 Maggi E, Marchesi E, Ravetta V, Martignoni A, Finardi G, Bellomo G. Presence of autoantibodies against oxidatively modified low density lipoprotein in essential hypertension: a biochemical signature of an enhanced in vivo low density lipoprotein oxidation.  J Hypertens. 1995;  13 129-138
• 24 Mironova M A, Klein R L, Virella G T, Lopes-Virella M F. Anti-modified LDL antibodies, LDL-containing immune complexes, and susceptibility of LDL to in vitro oxidation in patients with type 2 diabetes.  Diabetes. 2000;  49 1033-1041
• 25 Miyazaki H, Oh-ishi S, Ookawara T, Kizaki T, Toshinai K, Ha S, Haga S, Ji L L, Ohno H. Strenuous endurance training in humans reduces oxidative stress following exhausting exercise.  Eur J Appl Physiol. 2001;  84 1-6
• 26 Morrissey P A, Sheehy P J. Optimal nutrition: vitamin E.  Proc Nutr Soc. 1999;  58 459-468
• 27 Packer L. Oxidants, antioxidant nutrients and the athlete.  J Sports Sci. 1997;  15 353-363
• 28 Pincemail J, Deby C, Camus G, Pirnay F, Bouchez R, Masssaux L, Goutier R. Tocopherol mobilization during intensive exercise.  Eur J Appl Physiol. 1988;  57 189-191
• 29 Pincemail J, Lecomte J, Castiau J P, Collard E, Vasankari T, Cheramy-Bien J P, Limet T R, Defraigne J O. Evaluation of autoantibodies against oxidized LDL and antioxidant status in top soccer and basketball players after 4 months of competition.  Free Rad Biol Med. 2000;  28 559-565
• 30 Pincemail J, Siquet J, Chapelle J P, Cheramy-Bien J P, Paulissen G, Chantillon A M, Christiaens G, Gielen J, Limet R, Defraigne J O. Determination of concentrations of antioxidants, antibodies against oxidized LDL, and homocysteine in a population sample from Liège.  Ann Biol Clin (Paris). 2000;  58 177-185
• 31 Princen H MG, van Duyvenvoorde W, Buytenhek R, van der Laarse A, van Poppel G, Gevers Leuven J A, van Hinsbergh V WM. Supplementation with low doses of vitamin E protects LDL from lipid peroxidation in men and women.  Arterioscler Thromb Vasc Biol. 1995;  15 325-333
• 32 Sánchez-Quesada J L, Ortega H, Payés-Romero A, Serrat-Serrat J, Gonzáles-Sastre F, Lasunción M A, Ordónez-Llanos J. LDL from aerobically-trained subjects shows higher resistance to oxidative modification than LDL from sedentary subjects.  Atherosclerosis. 1997;  132 207-213
• 33 Schippinger G, Wonisch W, Abuja P M, Fankhauser F, Winklhofer-Roob B M, Halwachs G. Lipid peroxidation and antioxidant status in professional American football players during competition.  Eur J Clin Invest. 2002;  32 686-692
• 34 Seccia M, Albano E, Maggi E, Bellomo G. Circulating antibodies recognizing peroxidase-oxidized low density lipoprotein.  Arterioscler Tromb Vasc Biol. 1997;  17 134-140
• 35 Sen C K. Oxidants and antioxidants in exercise.  J Appl Physiol. 1995;  79 675-686
• 36 Shoji T, Fukumoto M, Kimoto E, Shinohara K, Emoto M, Tahara H, Koyama H, Ishimura E, Nakatani T, Miki T, Tsujimoto Y, Tabata T, Nishizawa Y. Antibody to oxidized low-density lipoprotein and cardiovascular mortality in end-stage renal disease.  Kidney Int. 2002;  62 2230-2237
• 37 Sobczak A, Skop B, Kula B. Simultaneous determination of serum retinol and alpha- and gamma-tocopherol levels in type II diabetic patients using high-performance liquid chromatography with fluorescence detection.  J Chromatogr B Biomed Sci Appl. 1999;  730 265-271
• 38 Steinberg D. Low density lipoprotein oxidation and its pathological significance.  J Biol Chem. 1997;  272 20963-20966
• 39 Steinerová A, Racek J, Stožický F, Zima T, Fialová L, Lapin A. Antibodies against oxidized LDL - theory and clinical use.  Physiol Rev. 2001;  50 131-141
• 40 Svensson M B, Ekblom B, Cotgreave I A, Norman B, Sjöberg B, Ekblom Ö, Sjödin B, Sjödin A. Adaptive stress response of glutathione and uric acid metabolism in man following controlled exercise and diet.  Acta Physiol Scand. 2002;  176 43-56
• 41 Szasz G, Gruber W, Bernt E. Creatine kinase in serum: 1. Determination of optimum reaction conditions.  Clin Chem. 1976;  22 650-656
• 42 Tatzber F, Esterbauer H. Autoantibodies to oxidized low-density lipoprotein. Bellomo G., Finardi E., Maggi E., Rice-Evans C Free Radicals IX. London; Richelieu Press 1995: 245-262
• 43 Van Tits L J, de Waart F, Hak-Lemmers H L, van Heijst P, de Graaf J, Demacker P N, Stalenhoef A F. Effects of alpha-tocopherol on superoxide production and plasma intercellular adhesion molecule-1 and antibodies to oxidized LDL in chronic smokers.  Free Radic Biol Med. 2001;  30 1122-1129
• 44 Vasankari T J, Kujala U M, Vasankari T M, Ahotupa M. Reduced oxidized LDL levels after a 10-month exercise programme.  Med Sci Sports Exerc. 1998;  30 1496-1501
• 45 Waring W S, Convery A, Mishra V, Shenkin A, Webb D J, Maxwell S RJ. .  Uric acid reduces exercise-induced oxidative stress in healthy adults Clin Sci. 2003;  105 425-430
• 46 Wetzstein C J, Shern-Brewer R A, Santanam N, Green N R, White-Welkley J E, Parthasarthy S. Does acute exercise affect the susceptibility of low density lipoprotein to oxidation?.  Free Radic Biol Med. 1998;  24 679-682
• 47 Winklhopfer-Roob B M, van't Hof M A, Shmerling D H. Reference values for plasma concentrations of vitamin E and A and carotenoids in a Swiss population from infancy to adulthood, adjusted for seasonal influences.  Clin Chem. 1997;  43 146-153