Impact of Cytokine- and FasL-induced Apoptosis in the β-Cell Line NIT-1

Abstract

Cytokine- and FasL-induced pathways contribute to β-cell death in type 1 diabetes. It remains unclear, however, whether pro-apoptotic cyto-kines or FasL have more apoptotic impact. Cytokine- and FasL-induced apoptosis were simulated using IL-1β/IFN-γ, Super-FasLigand and the β-cell line NIT-1. The role of caspases was addressed using the general caspase inhibitor ZVAD. Exposure to IL-1β/IFN-γ induced NIT-1 cell death. FasL augmented cytokine-induced cell death accompanied by increased caspase-3 activation, DNA fragmentation, and chromatin condensation. However, FasL mediated comparable effects on the mitochondrial transmembrane potential (Δψ_m) and nitrite in cytokine- and untreated cells. The cytokine-induced sequence of apoptotic events was (1) Fas, nitrite, (2) Δψ_m, (3) DNA fragmentation, cell death, and (4) chromatin condensation. In the presence of FasL, cell death and chromatin condensation appeared earlier implicating a compression of the apoptotic time course. General caspase inhibition using ZVAD prevented cell death, Δψ_m, and DNA fragmentation; however, Fas expression and nitrite were increased. In conclusion, cytokines account for the major part of cell death induced by the simultaneously action of FasL + IL-1β/IFN-γ. Caspases are of central importance for β-cell death.

type 1 diabetes - pancreatic β cell - cytokine-induced apoptosis - FasL-induced apoptosis

References

• 1 Cnop M, Welsh N, Jonas JC, Jorns A, Lenzen S, Eizirik DL. Mechanisms of pancreatic {beta}-cell death in type 1 and type 2 diabetes: many differences, few similarities.  Diabetes. 2005;  54 ((Suppl 2)) S97-S107
• 2 Augstein P, Stephens LA, Allison J, Elefanty AG, Ekberg M, Kay TW, Harrison LC. Beta-cell apoptosis in an accelerated model of autoimmune diabetes.  Mol Med. 1998;  4 495-501
• 3 Moriwaki M, Itoh N, Miyagawa J, Yamamoto K, Imagawa A, Yamagata K, Iwahashi H, Nakajima H, Namba M, Nagata S, Hanafusa T, Matsuzawa Y. Fas and Fas ligand expression in inflamed islets in pancreas sections of patients with recent-onset Type I diabetes mellitus.  Diabetologia. 1999;  42 1332-1340
• 4 Eizirik DL, Kutlu B, Rasschaert J, Darville M, Cardozo AK. Use of microarray analysis to unveil transcription factor and gene networks contributing to Beta cell dysfunction and apoptosis.  Ann N Y Acad Sci. 2003;  1005 55-74
• 5 Cardozo AK, Kruhoffer M, Leeman R, Orntoft T, Eizirik DL. Identification of novel cytokine-induced genes in pancreatic beta-cells by high-density oligonucleotide arrays.  Diabetes. 2001;  50 909-920
• 6 Karlsen AE, Ronn SG, Lindberg K, Johannesen J, Galsgaard ED, Pociot F, Nielsen JH, Mandrup-Poulsen T, Nerup J, Billestrup N. Suppressor of cytokine signaling 3 (SOCS-3) protects beta -cells against interleukin-1beta – and interferon-gamma -mediated toxicity.  Proc Natl Acad Sci USA. 2001;  98 12191-12196
• 7 Seewaldt S, Thomas HE, Ejrnaes M, Christen U, Wolfe T, Rodrigo E, Coon B, Michelsen B, Kay TW, Herrath MG von. Virus-induced autoimmune diabetes: most beta-cells die through inflammatory cytokines and not perforin from autoreactive (anti-viral) cytotoxic T-lymphocytes.  Diabetes. 2000;  49 1801-1809
• 8 Thomas HE, Irawaty W, Darwiche R, Brodnicki TC, Santamaria P, Allison J, Kay TW. IL-1 receptor deficiency slows progression to diabetes in the NOD mouse.  Diabetes. 2004;  53 113-121
• 9 Liu D, Pavlovic D, Chen MC, Flodstrom M, Sandler S, Eizirik DL. Cyto-kines induce apoptosis in beta-cells isolated from mice lacking the inducible isoform of nitric oxide synthase (iNOS-/-).  Diabetes. 2000;  49 1116-1122
• 10 Zumsteg U, Frigerio S, Hollander GA. Nitric oxide production and Fas surface expression mediate two independent pathways of cytokine-induced murine beta-cell damage.  Diabetes. 2000;  49 39-47
• 11 Augstein P, Wachlin G, Berg S, Bahr J, Salzsieder C, Hehmke B, Heinke P, Salzsieder E. Surface and intracellular Fas expression associated with cytokine-induced apoptosis in rodent islet and insulinoma cells1.  J Mol Endocrinol. 2003;  30 163-171
• 12 Stassi G, Maria R De, Trucco G, Rudert W, Testi R, Galluzzo A, Giordano C, Trucco M. Nitric oxide primes pancreatic beta cells for Fas-mediated destruction in insulin-dependent diabetes mellitus.  J Exp Med. 1997;  186 1193-1200
• 13 Suarez-Pinzon WL, Power RF, Rabinovitch A. Fas ligand-mediated mechanisms are involved in autoimmune destruction of islet beta cells in non-obese diabetic mice.  Diabetologia. 2000;  43 1149-1156
• 14 Allison J, Thomas HE, Catterall T, Kay TW, Strasser A. Transgenic expression of dominant-negative Fas-associated death domain protein in beta cells protects against Fas ligand-induced apoptosis and reduces spontaneous diabetes in nonobese diabetic mice.  J Immunol. 2005;  175 293-301
• 15 Hamaguchi K, Gaskins HR, Leiter EH. NIT-1, a pancreatic beta-cell line established from a transgenic NOD/Lt mouse.  Diabetes. 1991;  40 842-849
• 16 Augstein P, Heinke P, Salzsieder E, Berg S, Rettig R, Salzsieder C, Harrison LC. Fas ligand down-regulates cytokine-induced Fas receptor expression on insulinoma (NIT-1), but not islet cells, from autoimmune nonobese diabetic mice.  Endocrinology. 2004;  145 2747-2752
• 17 Augstein P, Dunger A, Salzsieder C, Heinke P, Kubernath R, Bahr J, Fischer U, Rettig R, Salzsieder E. Cell surface trafficking of Fas in NIT-1 cells and dissection of surface and total Fas expression.  Biochem Biophys Res Commun. 2002;  290 443-451
• 18 Zamzami N, Metivier D, Kroemer G. Quantitation of mitochondrial transmembrane potential in cells and in isolated mitochondria.  Methods Enzymol. 2000;  322 208-213
• 19 Augstein P, Heinke P, Salzsieder E, Grimm R, Giebel J, Bahr J, Salzsieder C, Harrison LC. Dominance of cytokine-over FasL-induced impairment of the mitochondrial transmembrane potential (Ψm) in the pancreatic beta-cell line NIT 1.  Diabetes Vasc Dis Res. 2008;  5 198-204
• 20 Augstein P, Bahr J, Wachlin G, Heinke P, Berg S, Salzsieder E, Harrison LC. Cytokines activate caspase-3 in insulinoma cells of diabetes-prone NOD mice directly and via upregulation of Fas.  J Autoimmun. 2004;  23 301-309
• 21 Delaney CA, Pavlovic D, Hoorens A, Pipeleers DG, Eizirik DL. Cytokines induce deoxyribonucleic acid strand breaks and apoptosis in human pancreatic islet cells.  Endocrinology. 1997;  138 2610-2614
• 22 Chen M, Yang Z, Wu R, Nadler JL. Lisofylline, a novel antiinflammatory agent, protects pancreatic beta-cells from proinflammatory cytokine damage by promoting mitochondrial metabolism.  Endocrinology. 2002;  143 2341-2348
• 23 Riachy R, Vandewalle B, Kerr CJ, Moerman E, Sacchetti P, Lukowiak B, Gmyr V, Bouckenooghe T, Dubois M, Pattou F. 1,25-dihydroxyvitamin D3 protects RINm5F and human islet cells against cytokine-induced apoptosis: implication of the antiapoptotic protein A20.  Endocrinology. 2002;  143 4809-4819
• 24 Barbu AR, Akusjarvi G, Welsh N. Adenoviral-induced islet cell cytotoxicity is not counteracted by Bcl-2 overexpression.  Mol Med. 2002;  8 733-741
• 25 Barbu A, Welsh N, Saldeen J. Cytokine-induced apoptosis and necrosis are preceded by disruption of the mitochondrial membrane potential (Deltapsi(m)) in pancreatic RINm5F cells: prevention by Bcl-2.  Mol Cell Endocrinol. 2002;  190 75-82
• 26 Thomas HE, Darwiche R, Corbett JA, Kay TW. Evidence that beta cell death in the nonobese diabetic mouse is Fas independent.  J Immunol. 1999;  163 1562-1569
• 27 Yamada K, Takane-Gyotoku N, Yuan X, Ichikawa F, Inada C, Nonaka K. Mouse islet cell lysis mediated by interleukin-1-induced Fas.  Diabetologia. 1996;  39 1306-1312
• 28 Burkhardt BR, Lyle R, Qian K, Arnold AS, Cheng H, Atkinson MA, Zhang YC. Efficient delivery of siRNA into cytokine-stimulated insulinoma cells silences Fas expression and inhibits Fas-mediated apoptosis.  FEBS Lett. 2006;  580 553-560
• 29 Eizirik DL, Mandrup-Poulsen T. A choice of death – the signal-transduction of immune-mediated beta cell apoptosis.  Diabetologia. 2001;  44 2115-2133
• 30 Kharroubi I, Ladriere L, Cardozo AK, Dogusan Z, Cnop M, Eizirik DL. Free fatty acids and cytokines induce pancreatic beta-cell apoptosis by different mechanisms: role of nuclear factor-kappaB and endoplasmic reticulum stress.  Endocrinology. 2004;  145 5087-5096
• 31 Ly JD, Grubb DR, Lawen A. The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update.  Apoptosis. 2003;  8 115-128
• 32 Launay S, Hermine O, Fontenay M, Kroemer G, Solary E, Garrido C. Vital functions for lethal caspases 29.  Oncogene. 2005;  24 5137-5148

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