ERYTHROCYTE DAMAGE AND LEUKOCYTE ACTIVATION AS POTENTIAL RISK MARKERS FOR CARDIOVASCULAR EVENTS

Abstract
The traditional lipid risk factors can only predict some of the cardiovascular events. Our recent work has focused on new potential biological markers of risk, namely erythrocyte damage and leukocyte activation. We have studied these potential markers in patients at risk for cardiovascular events, in myocardial infarction and ischemic stroke cases and in adolescents practising moderate and high competition physical exercise. Some in vitro studies were also performed to strengthen the value of the proposed markers. We found a rise in leukocytes which seem to be activated, as shown by the increase in their activation products, elastase and lactoferrin. This activation seems to impose erythrocyte damage and premature aging, as suggested by the increase in membrane bound hemoglobin and by a different band 3 profile. The in vitro studies showed that the modifications imposed to band 3 profile by increasing concentrations of activated neutrophils and elastase were correlated, and were similar to those presented by older erythrocytes. Our data suggest that plasma levels of elastase and lactoferrin, together with levels of erythrocyte membrane bound hemoglobin and band 3 profile, warrants further studies as potential new markers of risk for cardiovascular events.

Bibliografía del artículo

1. Sá P, Dias JÁ, Pereira Miguel JM. Ischaemic heart disease (IHD) and cerebrovascular disease (CVD) mortality evolution in Portugal, 1980-1989. Acta Méd Port, 1994, 7 : 71-81.
2. Marmot M. In Atherosclerosis XII, Ed Stemme S Olsson AG, Elsevier Science. Epidemiology and mechanisms of atherosclerosis: bringing the perspectives together. 2000, 125-37.
3. Hansson GK. In Atherosclerosis XII, Ed Stemme S Olsson AG, Elsevier Science. Inflammation, injury and atherosclerosis – the Russel Ross memorial lecture. 2000, 53-66.
4. Lucas AD, Greaves DR. Atherosclerosis: role of chemokines and macrophages. Exp Rev Mol Med, 2001, 5 : 1-18.
5. Davies PF, Barbee KA, Lal R, Robotewskyj A, Griem ML. Hemodynamics and atherogenesis. Endothelial surface dynamics in flow signal transduction. In Atherosclerosis III Recent Research Advances in Atherosclerosis. Numano F, Wissler RW Eds. Ann NY Acad Sci, 1995; 748:86-103.
6. Lowe GDO. Blood viscosity cardiovascular risk. Curr Opin Lipidol, 1993; 4 : 283-287.
7. Cicco G, Pirelli A. Red blood cell (RBC) deformability, RBC aggregability and tissue oxygenation in hypertension. Clin Hemorheol Circ 1999, 21 : 169-177.
8. Schleimer RP, Rutledge BK. Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin and tumor-promoting phobol esters. J Immunol 1986, 136 : 649.
9. Hoffman M, Blum A, Baruch R, Kaplan E, Benjamin M. Leukocytes and coronary artery disease. Atherosclerosis, 2004; 172 : 1-6.
10. Borregaard N, Lollike K, Kjeldsen L, Sengelov H, Bastholm L, Nielsen MH, Bainton DF. Human neutrophil granules and secretory vesicles. Eur J Haematol, 1993, 51 : 187-198.
11. Lutz HU. Erythrocyte clearance blood cell biochemistry. In: Harris JR, editor, Erythroid cells. New York : Plenum, 1990, 81-120.
12. Brovelli A, Castellana MA, Minetti G, Piccinini G, Seppi C, Renzis MR, Balduini C. Conformational changes and oxidation of membrane proteins in senescent human erythrocytes. In: Magnani M, De Flora A, editors. Red blood cell aging. New York: plenum, 1991, 59-73.
13. Kay MMB, Marchalonis JJ, Schluter SF, Bosman G. Human erythrocyte aging: cellular and molecular biology. Transf Med Rev, 1991, 3:173-95.
14. Bosch FH, Were JM, Scipper L, Roerdinkholder-Stoelwinder B, Huls T, Willekens FLA, Wichers G, Halie MR. Determinants of red blood cell deformability in relation to cell age. Eur J Haematol 1994, 52 : 35-41.
15. Kay MMB. Role of physiologic autoantibody in the removal of senescent human red cells. J Supramol Struct 1978, 9:555-67.
16. Kay MMB, Goodman SR, Sorensen K, Whitfield CF, Wong P, Zaki L, Rudloff V. Senescent cell antigen is immunologically related to band 3. Proc Natl Acad Sci USA 1983, 80:1631-5.
17. Kay MMB. Localization of senescent cell antigen on band 3. Proc Natl Acad Sci USA, 1984, 81:5753-7.
18. Kay MMB, Flowers N, Goodman SR, Bosman G. Alterations in membrane protein band 3 associated with accelerated erythrocyte aging. Proc Natl Acad Sci USA, 1989, 816:5834-7.
19. Lutz A. Naturally occurring anti-band 3 antibodies. Transf Med Rev, 1992, 3:201-211.
20. Bruce LJ, Tanner MJA. Erythroid band 3 variants and disease. Ballière\'s Clin Haematol, 1999, 12(4):637-654.
21. Low PS. Role of hemoglobin denaturation and band 3 clustering in intiating red cell removal. In: Magnani M, De Flora A, editors. Red blood cell aging. New York: Plenum, 1991, 173-183.
22. Lutz HU, Bussolino F, Flepp R, Fasler S, Stammler P, Kazatchkine MD, Arese P. Naturally occurring anti-band 3 antibodies and complement activation together mediate phagocytosis of oxidatively stressed human erythrocytes. Proc Natl Acad Sci, 1987, 84 : 7368-7372.
23. Weiss SJ. Neutrophil-mediated methemoglobin formation in the erythrocyte. J Biol Chem, 1982, 257:2947-53.
24. Claster S, Chiu D, Quintanilha A, Lubin B. Neutrophils mediate lipid peroxidation in human red cells. Blood, 1984, 64:1079-84.
25. Vercellotti GM, van Asbeck BS, Jacob HS. Oxygen radical-induced erythrocyte hemolysis by neutrophils. Critical role of iron and lactoferrin. J Clin Ivest, 1985, 76 : 956-962.
26. Claster S, Quintanilha A, Schott MA, Chiu D, Lubin B. Human neutrophil elastase mediate red cells membrane damage. Blood, 1986, 68:92a-100a.
27. Uyesaka N, Hasegawa S, Ishioka N, Shio H, Schechter NA. Effects of superoxide anions on red cell deformability and membrane proteins. Biorheol, 1992, 29:217-29.
28. Weiss DJ, Aird B, Murtaugh MP. Neutrpil-mediated immunoglobulin binding to erythrocytes involves proteolytic and oxidative injury. J Leukocyte Biol, 1992, 29 : 217-229.
29. Santos-Silva A, Castro EMB, Teixeira NA, Guerra FC, Quintanilha A. Altered erythrocyte band 3 profile as a marker in patients at risk for cardiovascular disease. Atherosclerosis, 1995, 116:199-209.
30. Santos-Silva A, Castro EMB, Teixeira NA, Guerra FC, Quintanilha A. Erythrocyte membrane band 3 profile imposed by cellular aging,by activated neutrophils and by neutrophilic elastase. Clin Chim Acta, 1998, 275:185-196.
31. Thompson PD, Buchner D, Pina IL, Balady GJ, Williams MA, Marcus BH, Berra K, Blair SN, Costa F, Franklin B, Fletcher F, Gordon NF, Pate RR, Rodriguez BL, Yancey AK, Wenger NK. Exercise and physical activity in the prevention and treatment of arteriosclerotic disease. Circulation, 2003, 107 : 3109-3116.
32. Stefanick ML, Mackey S, Sheehan M, Ellsworth N, Haskell WL, Wood PD. Effects of diet and exercise in men and postmenopausal women with low levels of HDL cholesterol and high levels of LDL cholesterol. N Engl J Med, 1998, 339 : 12-20.
33. Powers SK, Lennon SL, Quindry J, Mehta JL. Exercise and cardioprotection. Curr Opin Cardiol, 2002, 17(5) : 495-502.
34. Sánchez-Quesada JL, Homs- Serradesanferm R, Serat-Serrat J, Serra-Grima JR, González-Sastre F, Ordonéz-Llanos J et al.. Increase of LDL susceptibility to oxidation occurring after intense, long duration aerobic exercise. Atherosclerosis, 1995, 118 : 297- .
35. Cooper Ce, Vollaard NB, Choueiri T, Wilson MT. Exercise, free radicals and oxidative stress. Biochem Soc Trans, 2002, 30(2) : 280-285.
36. Banerjee AK, Mandal A, Chanda D, Chakraborti S. Oxidant, antioxidant and physical exercise. Mol Cell Biochem, 2003, 253(1-2) : 307-312.
37. Santos-Silva A, Rebelo I, Castro EMB, Belo L, Guerra A, Rego C, Quintanilha A. Leukocyte activation, erythrocyte damage, lipid profile and oxidative stress imposed by high competition physical exercise in adolescents. Clin Chim Acta, 2001, 306:119-126.
38. Santos-Silva A, Rebelo I, Castro EMB, Belo L, Catarino C, Monteiro I, Almeida MD, Quintanilha A. Erythrocyte damage and leukocyte activation in ischemic stroke. Clin Chim Acta, 2002, 320:29-35.
39. Levenson J, Del-Pino M, Simon A. Blood and arterial wall rheology and cardiovascular risk factors. J Mal Vasc, 2000, 25(4):237-240.
40. Connor J, Pak CC, Schroit AJ. Exposure of phosphatidylserine in the outer leaflet of human red blood cells. Relationship to cell density, cell age, and clearance by mononuclear cells. J Biol Chem, 1994, 269(4):2399-2404.