High Susceptibility Of Activated Lymphocytes To Oxidative Stress-induced Cell Death

The present study provides evidence that activated spleen lymphocytes from Walker 256 tumor bearing rats are more susceptible than controls to iert-butyl hydroperoxide (t-BOOH)-induced necrotic cell death in vitro. The iron chelator and antioxidant deferoxamine, the intracellular Ca2+ chelator BAPTA, the L-type Ca2+ channel antagonist nifedipine or the mitochondrial permeability transition inhibitor cyclosporin A, but not the calcineurin inhibitor FK-506, render control and activated lymphocytes equally resistant to the toxic effects of t-BOOH. Incubation of activated lymphocytes in the presence of t-BOOH resulted in a cyclosporin A-sensitive decrease in mitochondrial membrane potential. These results indicate that the higher cytosolic Ca 2+ level in activated lymphocytes increases their susceptibility to oxidative stress-induced cell death in a mechanism involving the participation of mitochondrial permeability transition.801137148ABE, K., SAITO, H., Characterization of t-butyl hydroperoxide toxicity in cultured rat cortical neurones and astrocytes (1998) Pharmacol Toxicol, 83, pp. 40-46ARNOLD, R., BRENNER, D., BECKER, M., FREY, C.R., KRAMMER, P.H., How T lymphocytes switch between life and death (2006) Eur J Immunol, 36, pp. 1654-1658BARTESAGHI, S., TRUJILLO, M., DENICOLA, A., FOLKES, L., WARDMAN, P., RADI, R., Reactions of desferrioxamine with peroxynitrite-derived carbonate and nitrogen dioxide radicals (2004) Free Radic Biol Med, 36, pp. 471-483BARTOLI, G.M., PICCIONI, E., AGOSTARA, G., CALVIELLO, G., PALOZZA, P., Different mechanisms of tert-butyl hydroperoxide-induced lethal injury in normal and tumor thymocytes (1994) Arch Biochem Biophys, 312, pp. 81-87BERNARDES, C.F., PEREIRA, DA SILVA, L., VERCESI, A.E., t-Butylhydroperoxide-induced Ca2+ efflux from liver mitochondria in the presence of physiological concentrations of Mg2+ and ATP (1986) Biochim Biophys Acta, 850, pp. 41-48BOYUM, A., Isolation of lymphocytes, granulocytes and macrophages (1976) Scand J Immunol, (SUPPL. 5), pp. 9-15BRUMATTI, G., WEINLICH, R., CHEHAB, C.F., YON, M., AMARANTE-MENDES, G.P., Comparison of the anti-apoptotic effects of Bcr-Abl, Bcl-2 and Bcl-x(L) following diverse apoptogenic stimuli (2003) FEBS Lett, 541, pp. 57-63BUTTKE, T.M., SANDSTROM, P.A., Redox regulation of programmed cell death in lymphocytes (1995) Free Radic Res, 22, pp. 389-397CAMPOS, C.B., DEGASPERI, G.R., PACIFICO, D.S., ALBERICI, L.C., CARREIRA, R.S., GUIMARÃES, F., CASTILHO, R.F., VERCESI, A.E., Ibuprofen-induced Walker 256 tumor cell death: Cytochrome c release from functional mitochondria and enhancement by calcineurin inhibition (2004) Biochem Pharmacol, 68, pp. 2197-2206CASTILHO, R.F., KOWALTOWSKI, A.J., MEINICKE, A.R., BECHARA, E.J., VERCESI, A.E., Permeabilization of the inner mitochondrial membrane by Ca2+ ions is stimulated by t-butyl hydroperoxide and mediated by reactive oxygen species generated by mitochondria (1995) Free Radic Biol Med, 18, pp. 479-486CONKLIN, K.A., Chemotherapy-associated oxidative stress: Impact on chemotherapeutic effectiveness (2004) Integr Cancer Ther, 3, pp. 294-300CROMPTON, M., ELLINGER, H., COSTI, A., Inhibition by cyclosporin A of a Ca2+-dependent pore in heart mitochondria activated by inorganic phosphate and oxidative stress (1988) Biochem J, 255, pp. 357-360CROMPTON, M., The mitochondrial permeability transition pore and its role in cell death (1999) Biochem J, 341, pp. 233-249DALY, M.J., YOUNG, R.J., BRITNELL, S.L., NAYLER, W.G., The role of calcium in the toxic effects of tert-butyl hydroperoxide on adult rat cardiac myocytes (1991) J Mol Cell Cardiol, 23, pp. 1303-1312DEGASPERI, G.R., VELHO, J.A., ZECCHIN, K.G., SOUZA, C.T., VELLOSO, L.A., BORECKÝ, J., CASTILHO, R.F., VERCESI, A.E., Role of mitochondria in the immune response to cancer: A central role for Ca 2+ (2006) J Bioenerg Biomembr, 38, pp. 1-10DEGASPERI, G.R., ZECCHIN, K.G., BORECKY, J., CRUZ-HOFLING, M.A., CASTILHO, R.F., VELLOSO, L.A., GUIMARÃES, F., VERCESI, A.E., Verapamil-sensitive Ca2+ channel regulation of Th1-type proliferation of splenic lymphocytes induced by Walker 256 tumor development in rats (2006) Eur J Pharmacol, 549, pp. 179-184DOROSHOW, J.H., Anthracycline antibiotic-stimulated superoxide, hydrogen peroxide, and hydroxyl radical production by NADH dehydrogenase (1983) Cancer Res, 43, pp. 4543-4551FESKE, S., Calcium signalling in lymphocyte activation and disease (2007) Nat Rev Immunol, 7, pp. 690-702FRIBERG, H., FERRAND-DRAKE, M., BENGTSSON, F., HALESTRAP, A.P., WIELOCH, T., Cyclosporin A, but not FK 506, protects mitochondria and neurons against hypoglycemic damage and implicates the mitochondrial permeability transition in cell death (1998) JNeurosci, 18, pp. 5151-5159GALAT, A., Peptidylprolyl cis/trans isomerases (immunophilins): Biological diversity - targets - functions (2003) Curr Top Med Chem, 3, pp. 1315-1347GIARDINI, C., LA NASA, G., CONTU, L., GALIMBERTI, M., POLCHI, P., ANGELUCCI, E., BARONCIANI, D., LUCARELLI, G., Desferrioxamine therapy induces clearance of iron deposits after bone marrow transplantation for thalassemia: Case report (1993) Bone Marrow Transplant, (SUPPL. 1), pp. 108-110GOLDSTEIN, S., CZAPSKI, G., Transition metal ions and oxygen radicals (1990) Int Rev Exp Pathol, 31, pp. 133-164GREEN, D.R., KROEMER, G., The pathophysiology of mitochondrial cell death (2004) Science, 305, pp. 626-629GREEN, D.R., REED, J.C., Mitochondria and apoptosis (1998) Science, 281, pp. 1309-1312GRIFFITHS, E.J., HALESTRAP, A.P., Further evidence that cyclosporin A protects mitochondria from calcium overload, by inhibiting a matrix peptidyl-prolyl cis-trans isomerase. 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Life and death in peripheral T cells. 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Reactive Oxygen Species Generation In Peripheral Blood Monocytes And Oxidized Ldl Are Increased In Hyperlipidemic Patients

Objectives: Experimental and in vitro evidences have established that reactive oxygen species (ROS) generated by vascular wall cells play a key role in atherogenesis. Here, we evaluated the rate of ROS generation by resting peripheral monocytes in naive hyperlipidemic subjects. Design and methods: Primary hypercholesterolemic, combined hyperlipidemic, and normolipidemic individuals were studied. ROS generation and the mitochondrial electrical transmembrane potential were estimated by flow cytometry. Plasma oxidized (ox) LDL levels and lipid profile were measured by ELISA and enzymatic colorimetric methods. Results: Both hyperlipidemic groups presented significantly higher rates of monocyte ROS generation and elevated plasma levels of ox-LDL. Combined hyperlipidemic subjects presented increased levels of small dense LDL and insulin. Significant positive correlations between monocyte ROS generation and ox-LDL concentrations were found in pooled data. Conclusions: These data provide evidence that ROS production by circulating monocytes from hyperlipidemic subjects may contribute to the systemic oxidative stress and possibly to atherogenesis. © 2009 Elsevier Inc. All rights reserved.421212221227Chisolm, G.M., Steinberg, D., The oxidative modification hypothesis of atherogenesis: an overview (2000) Free Radic. Biol. Med., 28, pp. 1815-2186Stocker, R., Keaney Jr., J.F., Role of oxidative modifications in atherosclerosis (2004) Physiol. Rev., 84, pp. 1381-1478Napoli, C., Oxidation of LDL, atherogenesis, and apoptosis (2003) Ann. N.Y. Acad. Sci., 1010, pp. 698-709Duvall, W.L., Endothelial dysfunction and antioxidants (2005) Mt. Sinai J. Med., 72, pp. 71-80Halliwell, B., Gutteridge, J.M.C., The importance of free radicals and catalytic metal ions in human diseases (1985) Mol. Aspects Med., 8, pp. 89-193Boveris, A., Mitochondrial production of superoxide radical and hydrogen peroxide (1977) Adv. Exp. Med. 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Tumor growth characteristics of the Walker 256 AR tumor, a regressive variant of the rat Walker 256 A tumor

The present study aimed at characterizing the subcutaneous development of the Walker 256 (W256) AR tumor, a regressive variant of the rat W256 A tumor. Wistar rats were injected subcutaneously with 4x10(6) W256 A or W256 AR tumor cells. The development of tumors was evaluated daily by percutaneous measurements. None of the W256 A tumors (n=20) regressed, but 62% of the W256 AR tumor-bearing rats (n=21) underwent complete tumor regression within 35 days. Continuous growth of AR tumors was characterized by an increase of the tumor growth rate from day 12, which reached values above 1.0 g/day, and were significantly higher (p<0.05) than those of the regressive AR tumors. Immunosuppression by irradiation before subcutaneous injection of AR cells completely abrogated tumor regression and was associated with severe metastatic dissemination. Daily evaluation of the tumor growth rate enabled the discrimination, in advance, between continuously growing tumors and those that regressed later on.<br>O objetivo neste estudo foi caracterizar o desenvolvimento subcutâneo do tumor de Walker 256 (W256) AR, uma variante regressiva do tumor de W256 A de rato. Ratos Wistar foram injetados com 4x10(6) células tumorais de W256 A ou W256 AR. O desenvolvimento tumoral foi avaliado diariamente. Nenhum dos tumores W256 A (n=20) regrediu, mas 62% dos ratos com tumor W256 AR apresentaram regressão completa dos tumores em até 35 dias. O crescimento contínuo dos tumores AR foi caracterizado pelo aumento da taxa de crescimento tumoral a partir do dia 12, alcançando valores maiores que 1,0g/dia, que foram significativamente superiores (p<0,05) aos valores de taxa de crescimento dos tumores regressivos AR. A imunossupressão por irradiação precedendo a injeção das células tumorais AR eliminou completamente a regressão tumoral e favoreceu disseminação metastática severa. Este estudo caracterizou o desenvolvimento do tumor de W256 AR em condições específicas, documentando a regressão espontânea deste tumor após a injeção subcutânea de altas doses de células tumorais em ratos Wistar. A avaliação diária da taxa de crescimento tumoral permite discriminar precocemente os tumores com crescimento continuo daqueles que são regressivos. A taxa de crescimento tumoral é um parâmetro útil para a avaliação dos animais experimentais, particularmente no período que precede a regressão dos tumores