Redox imbalance of red blood cells impacts T lymphocyte homeostasis: implication in carotid atherosclerosis

 

 Buy Article Permissions and Reprints

Summary

Oxidative stress and immune/inflammatory responses are key pathogenetic factors of atherosclerotic disease. In this contest, mechanisms that regulate survival and death of immune cells may be relevant. Previous studies have demonstrated that red blood cells (RBCs) are physiologically able to inhibit apoptosis and to promote proliferation of activated T lymphocytes from healthy subjects. The aim of the present study was to evaluate whether RBCs from patients with carotid atherosclerosis maintain their property to modulate T cell homeostasis. Peripheral blood lymphocytes (PBLs) obtained from healthy subjects were activated in vitro by phytohemagglutinin in the presence/absence of RBCs from patients with carotid atherosclerosis or of in vitro oxidised RBCs from healthy subjects. Levels of reactive oxygen species (ROS) and aging markers of RBCs as well as susceptibility to apoptosis of PBLs were evaluated by flow cytometry. PBL proliferation was evaluated by 3H-methyl-thymidine incorporation assay whereas secretion of cytokines, analysed in view of their key role in T cell function, was assessed by ELISA. Levels of ROS and phosphatidyl-serine externalisation, a sign of RBC aging, resulted significantly higher in RBCs from patients than in those from healthy subjects, whereas surface glycophorin A expression and reduced glutathione content did the opposite. Unlike RBCs obtained from healthy subjects, RBCs from patients and in vitro oxidised RBCs did not protect activated T lymphocytes from apoptosis. Hence, RBCs from patients with carotid atherosclerosis, probably due to their oxidative imbalance, impact T cell integrity and function. Our results suggest a new regulatory role for RBCs in atherosclerosis.

• References

• 1 Hulsmans M, Holvoet P. The vicious circle between oxidative stress and inflammation in atherosclerosis. J Cell Mol Med 2010; 14: 70-78.
  CrossrefPubMedGoogle Scholar
• 2 Ross R. Atherosclerosis. An Inflammatory Disease. New Engl J Med 1999; 340: 115-126.
  CrossrefPubMedGoogle Scholar
• 3 Hansson GK. Mechanisms of disease: inflammation, atherosclerosis, and coronary artery disease. New Engl J Med 2005; 352: 1685-1695.
  PubMedGoogle Scholar
• 4 Hansson GK, Libby P. The immune response in atherosclerosis: a double-edged sword. Nature 2006; 6: 508-519.
  PubMedGoogle Scholar
• 5 Badimon L, Storey RF, Vilahur G. Update on lipids, inflammation and atherothrombosis. Thromb Haemost 2011; 105: S34-42.6.
  Article in Thieme ConnectPubMedGoogle Scholar
• 6 Madamanchi NR, Vendrov A, Runge MS. Oxidative stress and vascular disease. Arterioscler Thromb Vasc Biol 2005; 25: 29-38.
  PubMedGoogle Scholar
• 7 Puddu P, Puddu GM, Cravero E. et al. The emerging role of cardiovascular risk factor-induced mitochondrial dysfunction in atherogenesis. J Biomed Sci 2009; 16: 112
  CrossrefPubMedGoogle Scholar
• 8 Del Principe D, Frega G, Savini I. et al. The plasma membrane redox system in human platelet functions and platelet-leukocyte interactions. Thromb Haemost 2009; 101: 284-289.
  Article in Thieme ConnectPubMedGoogle Scholar
• 9 Vink A, Schoneveld AH, Richard W. et al. Plaque burden, arterial remodeling and plaque vulnerability: determined by systemic factors?. J Am Coll Cardiol 2001; 38: 718-723.
  CrossrefPubMedGoogle Scholar
• 10 Robertson AK, Hansson GK. T cells in atherogenesis. For better or for worse?. Arterioscler Thromb Vasc Biol 2006; 26: 2421-2432.
  CrossrefPubMedGoogle Scholar
• 11 Dahl TB, Yndestad A, Skjelland M. et al. Increased expression of visfatin in macrophages of human unstable carotid and coronary atherosclerosis: possible role in inflammation and plaque destabilization. Circulation 2007; 115: 972-980.
  CrossrefPubMedGoogle Scholar
• 12 Ylä-Herttuala S, Bentzon JF, Daemen M. et al. Stabilisation of atherosclerotic plaques. Position Paper of the European Society of Cardiology (ESC) Working Group of Atherosclerosis and Vascular Biology. Thromb Haemost 2011; 106: 1-19.
  Article in Thieme ConnectPubMedGoogle Scholar
• 13 Frostegard J, Ulfgren AK, Nyberg P. et al. Cytokine expression in advanced human atherosclerotic plaques: dominance of proinflammatory (Th1) and macrophage-stimulating cytokines. Atherosclerosis 1999; 145: 33-43.
  CrossrefPubMedGoogle Scholar
• 14 de Boer OJ, van der Wal AC, Houtkamp MA. et al. Unstable atherosclerotic plaques contain T cells that respond to Chlamydia pneumoniae. Cardiovasc Res 2000; 48: 402-408.
  CrossrefPubMedGoogle Scholar
• 15 Hu H, Zhu L, Huang Z. et al. Platelets enhance lymphocyte adhesion and infiltration into arterial thrombus. Thromb Haemost 2010; 104: 1184-1192.
  Article in Thieme ConnectPubMedGoogle Scholar
• 16 Munn LL, Melder RJ, Jain RK. Role of erythrocytes in leukocyte-endothelial interactions: mathematical model and experimental validation. Biophys J 1996; 71: 466-478.
  CrossrefPubMedGoogle Scholar
• 17 Melder RJ, Yuan J, Munn LL, Jain RK. Erythrocytes enhance lymphocyte rolling and arrest in vivo. Microvasc Res 2000; 59: 316-322.
  CrossrefPubMedGoogle Scholar
• 18 Arosa FA, Pereira CF, Fonseca AM. Red blood cells as modulators of T cell growth and survival. Curr Pharm Des 2004; 10: 191-201.
  CrossrefPubMedGoogle Scholar
• 19 Fonseca AM, Porto G, Uchida K. et al. Red blood cells inhibit activation-induced cell death and oxidative stress in human peripheral blood T lymphocytes. Blood 2001; 97: 3152-3160.
  CrossrefPubMedGoogle Scholar
• 20 Matarrese P, Straface E, Pietraforte D. et al. Peroxynitrite induces senescence and apoptosis of red blood cells through the activation of aspartyl and cysteinyl proteases. FASEB J 2005; 19: 416-418.
  CrossrefPubMedGoogle Scholar
• 21 Businaro R, Profumo E, Tagliani A. et al. Heat-shock protein 90: A novel autoantigen in human carotid atherosclerosis. Atherosclerosis 2009; 207: 74-83.
  CrossrefPubMedGoogle Scholar
• 22 Kotani K, Koibuchi H, Miyamoto M. et al. Relationship between reactive oxygen metabolites and carotid intima-media thickness in subjects with hypercholesterolemia. Med Princ Pract 2010; 19: 496-498.
  CrossrefPubMedGoogle Scholar
• 23 Caprari P, Bozzi A, Malorni W. et al. Junctional sites of erythrocyte skeletal proteins are specific targets of tert-butylhydroperoxide oxidative damage. Chem Biol Interact 1995; 94: 243-258.
  CrossrefPubMedGoogle Scholar
• 24 Mandal D, Mazumder A, Das P. et al. Fas-, caspase 8-, and caspase 3-dependent signaling regulates the activity of the aminophospholipid translocase and phosphatidylserine externalization in human erythrocytes. J Biol Chem 2005; 280: 39460-39467.
  CrossrefPubMedGoogle Scholar
• 25 Minetti M, Agati L, Malorni W. The microenvironment can shift erythrocytes from a friendly to a harmful behavior: pathogenetic implications for vascular diseases. Cardiovasc Res 2007; 75: 21-28.
  CrossrefPubMedGoogle Scholar
• 26 Nishi K, Itabe H, Uno M. et al. Oxidized LDL in carotid plaques and plasma associates with plaque instability. Arterioscler Thromb Vasc Biol 2002; 22: 1649-1654.
  CrossrefPubMedGoogle Scholar
• 27 Kockx MM, Cromheeke KM, Knaapen MW. et al. Phagocytosis and macrophage activation associated with hemorrhagic microvessels in human atherosclerosis. Arterioscler Thromb Vasc Biol 2003; 23: 440-446.
  CrossrefPubMedGoogle Scholar
• 28 Antunes RF, Brandão C, Maia M. et al. Red blood cells release factors with growth and survival bioactivities for normal and leukemic T cells. Immunol Cell Biol 2011; 89: 111-121.
  CrossrefPubMedGoogle Scholar
• 29 Gersh KC, Nagaswami C, Weisel JW. Fibrin network structure and clot mechanical properties are altered by incorporation of erythrocytes. Thromb Haemost 2009; 102: 1169-1175.
  Article in Thieme ConnectPubMedGoogle Scholar
• 30 Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. New York: Oxford University Press; 1999
  Google Scholar
• 31 Ashfaq S, Abramson JL, Jones DP. et al. The relationship between plasma levels of oxidized and reduced thiols and early atherosclerosis in healthy adults. J Am Coll Cardiol 2006; 47: 1005-1011.
  CrossrefPubMedGoogle Scholar
• 32 Dumaswala UJ, Zhuo L, Jacobsen DW. et al. Protein and lipid oxidation of banked human erythrocytes: role of glutathione. Free Radic Biol Med 1999; 27: 1041-1049.
  CrossrefPubMedGoogle Scholar
• 33 Rotstein R, Landau T, Twig A. et al. The erythrocyte adhesiveness/aggregation test (EAAT). A new biomarker to reveal the presence of low grade subclinical smoldering inflammation in individuals with atherosclerotic risk factors. Atherosclerosis 2002; 165: 343-351.
  CrossrefPubMedGoogle Scholar
• 34 Ingelsson E, Pencina MJ, Tofler GH. et al. Multimarker approach to evaluate the incidence of the metabolic syndrome and longitudinal changes in metabolic risk factors: the Framingham Offspring Study. Circulation 2007; 116: 984-992.
  CrossrefPubMedGoogle Scholar
• 35 Poli D, Antonucci E, Grifoni E. et al. Gender differences in stroke risk of atrial fibrillation patients on oral anticoagulant treatment. Thromb Haemost 2009; 101: 938-942.
  Article in Thieme ConnectPubMedGoogle Scholar
• 36 Snyder LM, Fortier NL, Trainor J. et al. Effect of hydrogen peroxide exposure on normal human erythrocyte deformability, morphology, surface characteristics, and spectrin-hemoglobin cross-linking. J Clin Invest 1985; 76: 1971-1977.
  CrossrefPubMedGoogle Scholar
• 37 Giulivi C, Davies KJ. Mechanism of the formation and proteolytic release of H2O2-induced dityrosine and tyrosine oxidation products in hemoglobin and red blood cells. J Biol Chem 2001; 276: 24129-24136.
  CrossrefPubMedGoogle Scholar
• 38 Romero N, Denicola A, Radi R. Red blood cells in the metabolism of nitric oxide-derived peroxynitrite. IUBMB Life 2006; 58: 572-580.
  CrossrefPubMedGoogle Scholar
• 39 Minetti M, Pietraforte D, Straface E. et al. Red blood cells as a model to differentiate between direct and indirect oxidation pathways of peroxynitrite. Methods Enzimol 2008; 440: 253-272.
  PubMedGoogle Scholar
• 40 Kalechman Y, Herman S, Gafter U. et al. Enhancing effects of autologous erythrocytes on human or mouse cytokine secretion and IL-2R expression. Cell Immunol 1993; 148: 114-129.
  CrossrefPubMedGoogle Scholar
• 41 Virella G, Rugeles MT, Hyman B. et al. The interaction of CD2 with its LFA-3 ligand expressed by autologous erythrocytes results in enhancement of B cell responses. Cell Immunol 1988; 116: 308-319.
  CrossrefPubMedGoogle Scholar
• 42 Taleb S, Tedgui A, Mallat Z. Adaptive T cell immune responses and atherogenesis. Curr Opin Pharmacol 2010; 10: 197-202.
  CrossrefPubMedGoogle Scholar
• 43 Tedgui A, Mallat Z. Cytokines in atherosclerosis: pathogenic and regulatory pathways. Physiol Rev 2006; 86: 515-581.
  CrossrefPubMedGoogle Scholar
• 44 Gupta S, Pablo AM, Jiang XC. et al. IFNgamma potentiates atherosclerosis in apoE knock-out mice. J Clin Invest 1997; 99: 2752-2761.
  CrossrefPubMedGoogle Scholar
• 45 Whitman SC, Ravisankar P, Elam H. et al. Exogenous interferon-gamma enhances atherosclerosis in apolipoprotein ES/S mice. Am J Pathol 2000; 157: 1819-1824.
  CrossrefPubMedGoogle Scholar
• 46 Whitman SC, Ravisankar P, Elam H. et al. T-bet deficiency reduces atherosclerosis and alters plaque antigenspecific immune responses. Proc Natl Acad Sci USA 2005; 102: 1596-1601.
  CrossrefPubMedGoogle Scholar
• 47 Schrijvers DM, De Meyer GR, Kockx MM. et al. Phagocytosis of apoptotic cells by macrophages is impaired in atherosclerosis. Arterioscler Thromb Vasc Biol 2005; 25: 1256-1261.
  CrossrefPubMedGoogle Scholar
• 48 Tedgui A, Mallat Z. Apoptosis as a determinant of atherothrombosis. Thromb Haemost 2001; 86: 420-426.
  Article in Thieme ConnectPubMedGoogle Scholar
• 49 Kockx MM, Knaapen MW. The role of apoptosis in vascular disease. J Pathol 2000; 190: 267-280.
  CrossrefPubMedGoogle Scholar
• 50 Bjorkerud S, Bjorkerud B. Apoptosis is abundant in human atherosclerotic lesions, especially in inflammatory cells (macrophages and T cells), and may contribute to the accumulation of gruel and plaque instability. Am J Pathol 1996; 149: 367-380.
  PubMedGoogle Scholar

• Supplementary Material