Abstract
Our earlier studies have shown that calorie restriction and n-3 fatty acids inhibit autoimmune disease and prolong life span. Experiments were designed to study the alteration of apoptosis and its mediators in B/W mice fed either n-6 fatty acids [5% corn oil (CO)] or n-3 fatty acids [5% fish oil (FO)] and either allowed access to the diet ad libitum (AL) or restricted in caloric intake by 40% (CR), from 4 weeks of age. At 4 months (young) and 9 months (old) mice were killed, splenic lymphocytes were isolated, and apoptosis was measured with Annexin V and PI staining. Apoptosis was decreased in splenic lymphocytes from both young and old CR mice compared to their respective AL-fed control groups regardless of fat source. Increasing apoptosis with age was observed in CO/AL, CO/CR, and FO/AL mice which correlated closely with significantly higher cellular peroxides measured by flow cytometry using dichlorofluourescein diacetate (DCFH-DA), whereas in both CO/CR and FO/CR peroxide levels remained low in old mice. Furthermore, CR increased the proliferative response of splenic lymphocytes and decreased the Fas (CD95) and Fas-L protein expression in CD4+ lymphocytes from old mice. Higher levels of Fas and Fas-L expression were observed in old mice compared to young mice. Bcl-2 levels were elevated in both young and old CR groups compared to the respective AL groups. Calorie restriction prevented the loss of CD8 cells in old mice fed both the CO and the FO diet. In summary, CR resulted in decreased apoptosis accompanied by alterations in Fas, Fas-L, and Bcl-2 expression in old mice, increased life span, and delayed onset of kidney disease.
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REFERENCES
Talal N, Steinberg AD: The pathogenesis of autoimmunity in New Zealand black mice.Curr Top Microbiol Immunol 64:79–103, 1974
Fernandes G, Friend P, Yunis EJ, Good RA: Influence of dietary restriction on immunologic function and renal disease in NZB×NZWF1 mice.Proc Natl Acad Sci, USA 75:1500–1504, 1978
Jolly C, Muthukumar A, Reddy Avula C, Troyer D, Fernandes G: Life span is prolonged in food-restricted autoimmune-prone (NZB × NZW)F(1) mice fed a diet enriched with (n-3) fatty acids. J Nutr131:2753–2760,2001
Prickett JD, Robinson DR, Steinberg AD: Dietary enrichment with the PUFA eicosapentaenoic prevents proteinuria and prolongs survival in NZB×NZWF1 mice.J Clin Invest658:556–559,1981
Fernandes G, Bysani C, Venkatraman JT, Tomar V, Zhao W: Increased TGF-beta and decreased oncogene expression by omega-3 fatty acids in the spleen delays onset of autoimmune disease in B/W mice.J Immunol152:5979–5987,1994
Fernandes G, Yunis EJ, Good RA: Influence of diet on survival of mice.Proc Natl Acad Sci USA73:1279–1283,1976
Jolly CA, Muthukumar A, Reddy Avula CP, Fernandes G: Maintenance of NF-kappaB activation in T-lymphocytes and a naive T-cell population in autoimmune-prone (NZB/NZW)F(1) mice by feeding a food-restricted diet enriched with n-3 fatty acids.Cell Immunol 213:122–133,2001
Chandrasekar B, McGuff HS, Aufdermorte TB, Troyer DA, Talal N, Fernandes G: Effects of calorie restriction on transforming growth factor beta 1 and proinflammatory cytokines in murine Sjogren' syndrome.Clin Immunol Immunopathol76:291–296, 1995
Chandrasekar B, Fernandes G: Decreased pro-inflammatory cytokines and increased antioxidant enzyme gene expression by ω-3 lipids in murine lupus nephritis.Biochem Biophys Res Commun 200:893–898,1994
Venkatraman JT, Poomchai Angkeow MS, Fernandes G: Effects of food restriction on antioxidant defense system in exercised rats. Nutrition Research18:283–298,1998
Byun DS, Venkatraman JT, Yu BP, Fernandes G:Modulation of antioxidant activities and immune response by food restriction in aging Fischer-344 rats.Aging (Milano) 7:40–48,1995
Fernandes G, Chandrasekar B, Troyer DA, Venkatraman JT, Good RA: Dietary lipids and calorie restriction affect mammary tumor incidence and gene expression in mouse mammary tumor virus/v-Ha-ras transgenic mice.Proc Natl Acad Sci USA92:6494–6498, 1995
Buttke TM, Sandstrom PA:Oxidative stress as a mediator of apoptosis.Immunol Today15:7–10,1994
Nagata S, Golstein P: The Fas death factor.Science267:1449–1456,1995
Green DR, Scott DW: Activation-induced apoptosis in lymphocytes. Curr Opin Immunol6:476–487,1994
Golstein P, Ojcius DM, Young JD: Cell death mechanisms and the immune system.Immunol Rev121:29–65,1991
Szondy Z, Reichert U, Fesus L: Retinoic acids regulate apoptosis of T lymphocytes through an interplay between RAR and RXR receptors [see comments].Cell Death Differ5:4–10,1998
Nagata S: Apoptosis by death factor.Cell 88:355–365,1997
Watanabe-Fukunaga R, Brannan CI, Itoh N, Yonehara S, Copeland NG, Jenkins NA, Nagata S: The cDNA structure, expression and chromosomal assignment of the mouse Fas antigen.J Immunol 148:1274–1279,1992
Suda T, Takahashi T, Golstein P, Nagata S: Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family.Cell75:1169–1178,1993
Suda T, Nagata S: Purification and characterization of the Fasligand that induces apoptosis.J Exp Med 179:873–879,1994
Nagata S: Apoptosis regulated by a death factor and its receptor: Fas ligand and Fas.Philos Trans R Soc Lond B Biol Sci 345:281–287,1994
Watanabe-Fukunaga R, Brannan CI, Copeland NG, Jenkins NA, Nagata S: Lymphoproliferative disorder in mice explained by defects in fas antigen that mediates apoptosis.Nature 356:314–317,1992
Takahashi T, Tanaka M, Brannan CI, Jenkins NA, Copeland NG, Suda T, Nagata S: Generalized lymphoproliferative disease in mice, caused by a point mutation in the fas ligand.Cell 76:969–976,1994
Cheng J, Liu C, Yang P, Zhou T, Mountz JD: Increased lymphocyte apoptosis in Fas ligand transgenic mice.J Immunol159:674–684,1997
Boise LH, Gonzalez-Garcia M, Postema CE, Ding L, Lindsten T, Turka L, Mao X, Nunez G, Thompson CB: Bcl-x, a bcl-2 related gene that functions as a dominant regulator of apoptotic cell death. Cell 74:597–608,1993
Reed JC: Double identity for proteins of the Bcl-2 family.Nature 387:773–776,1997
Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ:Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75:241–251,1993
Kane DJ, Sarafian TA, Anton R, Hahn H, Gralla EB, Balentine JS, Ord T, Bredesen DE: Bcl-2 inhibition of neural death: Decreased generation of reactive oxygen species.Science 262:1274–1277, 1993
Hotchkiss RS, Swanson PE, Knudson CM, Chang KC, Cobb JP, Osborne DF, Zollner KM, Buchman TG, Korsmeyer SJ, Karl IE: Overexpression of Bcl-2 in transgenic mice decreases apoptosis and improves survival in sepsis.J Immunol162:4148–4156,1999
Van Parijs L, Biuckians A, Abbas AK:Functional roles of Fas and Bcl-2-regulated apoptosis of T lymphocytes.J Immunol160:2065–2071,1998
Cory S: Regulation of lymphocyte survival by the bcl-2 gene family.Annu Rev Immunol 13:513–543,1995
Singer GG, Abbas AK: The fas antigen is involved in peripheral but not thymic deletion of T lymphocytes in T cell receptor transgenic mice.Immunity1:365–371,1994
Troyer DA, Chandrasekar B, Barnes JL, Fernandes G:Calorie restriction decreases platelet-derived growth factor (PDGF)-A and thrombin receptor mRNA expression in autoimmune murine lupus nephritis.Clin Exp Immunol108:58–62,1997
Troyer DA, Chandrasekar B, Thinnes T, Stone A, Loskutoff DA, Fernandes G: Effects of energy intake on type I plasminogen activator inhibitor levels in glomeruli of lupus-prone B/W mice. Am J Pathol 146:111–120,1995
Avula CP, Zaman AK, Lawrence RA, Fernandes G: Induction of apoptosis and apoptotic mediators in Balb/C splenic lymphocytes by dietary n-3 and n-6 fatty acids.Lipids 34:921–927,1999
Darzynkiewicz Z, Bruno S, Del Bino G, Gorzyca W, Holz MA, Lassota P, Traganos F: Features of apoptotic cells measured by flow cytometry.Cytometry13:795–808,1992
Avula CP, Fernandes G:Modulation of antioxidant enzymes and apoptosis in mice by dietary lipids and treadmill exercise.J Clin Immunol 19:35–44,1999
Bass DA, Parce JW, Dechatelet LR, Szejda P, Seeds MC, Thomas M:Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation.J Immunol 130:1910–1917,1983
Lim BO, Jolly CA, Zaman K, Fernandes G: Dietary (n-6) and (n-3) fatty acids and energy restriction modulate mesenteric lymph node lymphocyte function in autoimmune-prone (NZB × NZW)F1 mice.J Nutr130:1657–1664,2000
Adachi Y, Inaba M, Sugihara A, Koshiji M, Sugiura K, Amoh Y, Mori S, Kamiya T, Genba H, Ikehara S: Effects of administration of monoclonal antibodies (anti-CD4 or anti-CD8) on the development of autoimmune diseases in (NZW × BXSB)F1 mice. Immunobiology198:451–464,1998
Wolfe JT, Ross D, Cohen GM:A role for metals and free radicals in the induction of apoptosis in thymocytes.FEBS Letts352:58–62,1994
Duthie SJ, Ma A, Ross MA, Collins AR: Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res56:1291–1295,1996
Reddy Avula CP, Fernandes G:Effect of dietary vitamin E on apoptosis and proliferation of murine splenocytes.Nutr Res20: 225–236,2000
Djuric Z, Kritschevsky D: Modulation of oxidative DNA damage levels by dietary fat and calories.Mutat Res295:181–190,1993
Wolf NS, Penn PE, Jiang D, Fei RG, Pendergrass WR: Caloric restriction: conservation of in vivo cellular replicative capacity accompanies life-span extension in mice.Exp Cell Res217:317–323,1995
Sohal RS, Ku HH, Agarwal S, Forster MJ, Lal H: Oxidative damage, mitochondrial oxidant generation and antioxidant defenses during aging and in response to food restriction in the mouse.Mech Ag Develop74:121–133,1994
Kim JD, Yu BP, McCarter RJM, Lee SY, Herlihy JT:Exercise and diet modulate cardiac lipid peroxidation and antioxidant defenses. Free Radical Biol Med20:83–88,1996
Ogawa N, Dang H, Kong L, Anaya JM, Liu GT, Talal N: Lymphocyte apoptosis and apoptosis-associated gene expression in Sjögren' syndrome.Arthritis Rheum39:1875–1885,1996
Wu J, Wilson J, He J, Xiang L, Schur PH, Mountz JD: Fas ligand mutation in a patient with systemic lupus erythematosus and lymphoproliferative disease.J Clin Invest98:1107–1113,1996
Pensati L, Costanzo A, Ianni A, Accapezzato D, Iorio R, Natoli G, Nisini R, Almerighi C, Balsano C, Vajro P, Vegnente A, Levrero M:Fas/Apo1 mutations and autoimmune lymphoproliferative syndrome in a patient with type 2 autoimmune hepatitis [see comments]. Gastroenterology 113:1384–1389,1997
Dianzani U, Bragardo M, DiFranco D, Alliaudi C, Scagni P, Buonfiglio D, Redoglia V, Bonissoni S, Correra A, Dianzani I, Ramenghi U: Deficiency of the Fas apoptosis pathway without Fas gene mutations in pediatric patients with autoimmunity/ lymphoproliferation.Blood89:2871–2879,1997
Zeher M, Szodoray P, Gyimesi E, Szondy Z: Correlation of increased susceptibility to apoptosis of CD4+ T cells with lymphocyte activation and activity of disease in patients with primary Sjogren' syndrome.Arthritis Rheum42:1673–1681,1999
Kovacs B, Liossis SN, Dennis GJ, Tsokos GC:Increased expression of functional Fas-ligand in activated T cells from patients with systemic lupus erythematosus.Autoimmunity25:213–221,1997
Emlen W, Niebur J, Kadera R: Accelerated in vitro apoptosis of lymphocytes from patients with systemic lupus erythematosus. J Immunol 152:3685–3692,1994
Lorenz HM, Grunke M, Hieronymus T, Herrmann M, Kuhnel A, Manger B, Kalden JR:In vitro apoptosis and expression of apoptosis-related molecules in lymphocytes from patients with systemic lupus erythematosus and other autoimmune diseases [see comments].Arthritis Rheum40:306–317,1997
Kawabe Y, Ochi A:Programmed cell death and extrathymic reduction of Vβ8+ CD4+; T-cells in mice tolerant to Staphylococcus aureus enterotoxin B.Nature349:245–248,1991
Nakajima A, Hirai H, Kayagaki N, Yoshino S, Hirose S, Yagita H, Okumura K:Treatment of lupus in NZB/W F1 mice with monoclonal antibody against Fas ligand.J Autoimmun14:151–157, 2000
Tan EM: Autoimmunity and apoptosis.J Exp Med 179:1083–1086,1994
Itoh N, Tsujimoto Y, Nagata S:Effect of bcl-2 on Fas antigenmediated cell death.J Immunol151:621–627, 1993
Kawahara A, Kobayashi T, Nagata S:Inhibition of Fas-induced apoptosis by Bcl-2.Oncogene17:2549–2554,1998
Kamada S, Shimono A, Shinto Y, Tsujimura T, Takahashi T, Noda T, Kitamura Y, Kondoh H, Tsujimoto Y: bcl-2 deficiency in mice leads to pleiotropic abnormalities: accelerated lymphoid cell death in thymus and spleen, polycystic kidney, hair hypopigmentation, and distorted small intestine.Cancer Res55:354–359,1995
Aggarwal S, Gollapudi S, Gupta S: Increased TNF-alpha-induced apoptosis in lymphocytes from aged humans: changes in TNFalpha receptor expression and activation of caspases. J Immunol 162:2154–2161,1999
Shinohara S, Sawada T, Nishioka Y, Tohma S, Kisaki T, Inoue T, Ando K, Ikeda M, Fujii H, Ito K: Differential expression of Fas antigen and Bcl-2 protein on CD4+ T cells, CD8+ T cells, and monocytes.Cell Immunol 163:303–308,1995
Petit PX, Susin SA, Zamzami N, Mignotte B, Kroemer G: Mitochondria and programmed cell death: Back to the future. FEBS Lett396:7–13,1996
Kruman I, Bruce-Keller AJ, Bredesen D, Waeg G, Mattson MP: Evidence that 4-hydroxynonenal mediates oxidative stress-induced neuronal apoptosis.J Neurosci17:5089–5100,1997
Makinodan T, Chang MP, Norman DC, Li SC:Vulnerability of T cells to aging. In Aging and Immune Response–Cellular and Humoral Aspects, E Goidl (ed). New York, Marcel Dekker, 1987, p 827
Thoman ML, Weigle WO: The cellular and subcellular bases of immunosenescence.Adv Immunol 46:221–261, 1989
Fernandes G, Venkatraman JT, Khare A, Horbach GJMJ, Friedrichs W: Modulation of gene expression in autoimmune disease and aging by food restriction and by dietary lipids.Proc Soc Exp Biol Med 193:16–22,1990
Lee CK, Klopp RG, Weindruch R, Prolla TA: Gene expression profile of aging and its retardation by caloric restriction.Science 285:1390–1393,1999
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Reddy Avula, C., Lawrence, R.A., Zaman, K. et al. Inhibition of Intracellular Peroxides and Apoptosis of Lymphocytes in Lupus-Prone B/W Mice by Dietary n-6 and n-3 Lipids with Calorie Restriction. J Clin Immunol 22, 206–219 (2002). https://doi.org/10.1023/A:1016088708457
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DOI: https://doi.org/10.1023/A:1016088708457