Advertisement

Diabetologia

, Volume 39, Issue 5, pp 530–536 | Cite as

Cytokine-induced apoptotic cell death in a mouse pancreatic beta-cell line: inhibition by Bcl-2

  • H. Iwahashi
  • T. Hanafusa
  • Y. Eguchi
  • H. Nakajima
  • J. Miyagawa
  • N. Itoh
  • K. Tomita
  • M. Namba
  • M. Kuwajima
  • T. Noguchi
  • Y. Tsujimoto
  • Y. Matsuzawa
Originals

Summary

Cytokines are thought to contribute to the induction of pancreatic beta-cell destruction in insulin-dependent diabetes mellitus. The molecular mechanisms that underlie beta-cell death were investigated by studying cytokine-induced cell death in beta-cell lines. A combination of three cytokines (interleukin-1Β, tumour necrosis factor-α, and interferon-γ) induced apoptotic cell death in the mouse pancreatic beta-cell line ΒTC1, as judged from the appearance of cells with hypodiploid nuclei and oligonucleosomal DNA fragmentation. The same treatment also induced apoptosis in the mouse pancreatic alpha-cell line αTC1 and the NOD/Lt mouse beta-cell line NIT-1, although to a lesser extent than in ΒTC1 cells. The abundance of endogenous Bcl-2 in ΒTC1 cells was lower than that in the other two cell lines. Overexpression of human Bcl-2 in ΒTC1 cells partially protected them from cytokine-induced cell death. These results suggest that apoptosis may be responsible, at least in part, for cytokine-induced beta-cell destruction and that Bcl-2 prevents apoptosis in pancreatic islet cells.

Keywords

Pancreatic beta cell Bcl-2 apoptosis cytokine interleukin-1 tumour necrosis factor interferon-γ 

Abbreviations

IDDM

Insulin-dependent diabetes mellitus

IL

interleukin

TNF

tumour necrosis factor

IFN

interferon

FBS

fetal bovine serum

ATA

aurintricarboxylic acid

CHX

cycloheximide

PI

propidium iodide

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bottazzo GF, Dean BM, McNally JM, Mackay EH, Swift PGF, Gamble DR (1985) In situ characterization of autoimmune phenomena and expression of HLA molecules in the pancreas in diabetes insulitis. N Engl J Med 313: 353–360Google Scholar
  2. 2.
    Itoh N, Hanafusa T, Miyazaki A, et al. (1993) Mononuclear cell infiltration and its relation to the expression of major histocompatibility complex antigens and adhesion molecules in pancreas biopsy specimens from newly diagnosed insulin-dependent diabetes mellitus patients. J Clin Invest 92: 2313–2322Google Scholar
  3. 3.
    Pukel C, Baquerizo H, Rabinovitch A (1988) Destruction of rat islet cell monolayers by cytokines: synergistic interactions of interferon-γ, tumour necrosis factor, lymphotoxin, and interleukin-1. Diabetes 37: 133–136Google Scholar
  4. 4.
    Campbell IL, Iscalo A, Harrison LC (1988) IFN-γ and tumour necrosis factor-α: cytotoxicity to murine islets of Langerhans. J Immunol 141: 2325–2329Google Scholar
  5. 5.
    Wright SC, Kumar P, Tam AW, Shen N, Varma M, Larrick JW (1992) Apoptosis and DNA fragmentation precede TNF-induced cytolysis in U937 cells. J Cell Biochem 48: 344–355Google Scholar
  6. 6.
    Delaney CA, Green MH, Lowe JE, Green IC (1993) Endogenous nitric oxide induced by interleukin-1β in rat islets of Langerhans and HIT-T15 cells causes significant DNA damage as measured by the ‘comet’ assay. FEBS Lett 333: 291–295Google Scholar
  7. 7.
    Yamada K, Otabe S, Inada C, Takane N, Nonaka K (1993) Nitric oxide and nitric oxide synthase mRNA induction in mouse islet cells by interferon-γ plus tumour necrosis factor-α. Biochem Biophys Res Commun 197: 22–27Google Scholar
  8. 8.
    Suarez-Pinzon WL, Strynadka K, Schulz R, Rabinovitch A (1994) Mechanisms of cytokine-induced destruction of rat insulinoma cells: the role of nitric oxide. Endocrinology 134: 1006–1010Google Scholar
  9. 9.
    Albina JE, Cui S, Mateo RB, Reichner JS (1993) Nitric oxide-mediated apoptosis in murine peritoneal macrophages. J Immunol 150: 5080–5085Google Scholar
  10. 10.
    Rabinovitch A, Suarez-Pinzon WL, Shi Y, Morgan AR, Bleackley RC (1994) DNA fragmentation is an early event in cytokine-induced islet beta-cell destruction. Diabetologia 37: 733–738Google Scholar
  11. 11.
    Tsujimoto Y, Gorham J, Crossman J, Jaffe E, Croce CM (1985) The t(14;18) chromosome translocations involved in B-cell neoplasms result from mistakes in VDJ joining. Science 229: 1390–1393Google Scholar
  12. 12.
    Vaux DL, Cory S, Adams JM (1988) Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 335: 440–442Google Scholar
  13. 13.
    Garcia I, Martinou I, Tsujimoto Y, Martinou JC (1992) Prevention of programmed cell death of sympathetic neurons by the bcl-2 proto-oncogene. Science 258: 302–304Google Scholar
  14. 14.
    Nunez G, London L, Hockenbery D, Alexander M, McKearn JP (1990) Deregulated bcl-2 gene expression selectively prolongs survival of growth factor-deprived hemopoietic cell lines. J Immunol 144: 3602–3610Google Scholar
  15. 15.
    Strasser A, Whittingham S, Vaux DL, et al. (1991) Enforced BCL2 expression in B-lymphoid cells prolongs antibody responses and elicits autoimmune disease. Proc Natl Acad Sci USA 88: 8661–8665Google Scholar
  16. 16.
    Martinou JC, Dauphin DM, Staple JK, et al. (1994) Overexpression of BCL-2 in transgenic mice protects neurons from naturally occurring cell death and experimental ischemia. Neuron 13: 1017–1030Google Scholar
  17. 17.
    Veis DJ, Sorenson CM, Shutter JR, Korsmeyer SJ (1993) Bcl-2-deficient mice demonstrate fulminant lymphoid apoptosis, polycystic kidneys, and hypopigmented hair. Cell 75: 229–240Google Scholar
  18. 18.
    Nakayama K, Nakayama KI, Negishi I, Kuida K, Sawa H, Loh DY (1994) Targeted disruption of Bcl-2αΒ in mice: occurrence of gray hair, polycystic kidney disease, and lymphocytopenia. Proc Natl Acad Sci USA 91: 3700–3704Google Scholar
  19. 19.
    Kamada S, Shimono A, Shinto Y, et al. (1995) 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 Research 55: 354–359Google Scholar
  20. 20.
    Hockenbery DM, Zutter M, Hickey W, Nahm M, Korsmeyer SJ (1991) Bcl-2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci USA 88: 6961–6965Google Scholar
  21. 21.
    LeBrun DP, Warnke RA, Cleary ML (1993) Expression of bcl-2 in fetal tissues suggests a role in morphogenesis. Am J Pathol 142: 743–753Google Scholar
  22. 22.
    Efrat S, Linde S, Kofod H, et al. (1988) Beta-cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc Natl Acad Sci USA 85: 9037–9041Google Scholar
  23. 23.
    Powers AC, Efrat S, Mojsov S, Spector D, Habener JF, Hanahan D (1990) Proglucagon processing similar to normal islets in pancreatic α-like cell line derived from transgenic mouse tumor. Diabetes 39: 406–414Google Scholar
  24. 24.
    Hamaguchi K, Gaskins HR, Leiter EH (1991) NIT-1, a pancreatic Β-cell line established from a transgenic NOD/Lt mouse. Diabetes 40: 842–849Google Scholar
  25. 25.
    Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139: 271–279Google Scholar
  26. 26.
    Wyllie AH, Morris RG (1982) Hormone-induced cell death; purification and properties of thymocytes undergoing apoptosis after glucocorticoid treatment. Am J Pathol 109: 78–87Google Scholar
  27. 27.
    Borzillo GV, Endo K, Tsujimoto Y (1992) Bcl-2 confers growth and survival advantage to interleukin 7-dependent early pre-B cells which become factor independent by a multistep process in culture. Oncogene 7: 869–876Google Scholar
  28. 28.
    Wyllie AH (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68: 251–307Google Scholar
  29. 29.
    Baumgartner-Parzer SM, Wagner L, Pettermann M, Grillari J, Gessl A, Waldhausl W (1995) High-glucose triggered apoptosis in cultured endothelial cells. Diabetes 44: 1323–1327Google Scholar
  30. 30.
    Pender MP, Nguyen KB, McCombe PA, Kerr JFR (1991) Apoptosis in the nervous system in experimental allergic encephalomyelitis. J Neurol Sci 104: 81–87Google Scholar
  31. 31.
    Ankarcrona M, Dypbukt JM, Brune B, Nicotera P (1994) Interleukin-1Β-induced nitric oxide production activates apoptosis in pancreatic RINm5F cells. Exp Cell Res 213: 172–177Google Scholar
  32. 32.
    Kaneto H, Fujii J, Seo HG, et al. (1995) Apoptotic cell death triggered by nitric oxide in pancreatic Β-cells. Diabetes 44: 733–738Google Scholar
  33. 33.
    Boice LH, Gonzalez-Garcia M, Postema CE, et al. (1993) bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 74: 597–608Google Scholar
  34. 34.
    Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. Cell 74: 609–619Google Scholar
  35. 35.
    Takayama S, Sato T, Krajewski S, et al. (1995) Cloning and functional analysis of BAG-1: a novel Bcl-2-binding protein with anti-cell death activity. Cell 80: 279–284Google Scholar
  36. 36.
    Yang E, Zha J, Jockel J, Boice LH, Thompson CB, Korsmeyer SJ (1995) Bad, a heterodimeric partner for Bcl-xL and Bcl-2, displaces Bax and promotes cell death. Cell 80: 285–291Google Scholar
  37. 37.
    Farrow SN, White JHM, Martinou I, et al. (1995) Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19 K. Nature 374: 731–733Google Scholar
  38. 38.
    Chittenden T, Harrington EA, O'Connor R, et al. (1995) Induction of apoptosis by the Bcl-2 homologue Bak. Nature 374: 733–736Google Scholar
  39. 39.
    Kiefer MC, Brauer MJ, Powers VC, et al. (1995) Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak. Nature 374: 736–739Google Scholar
  40. 40.
    Harrison LC, Campbell IL, Colman PG, et al. (1990) Type 1 diabetes: immunology and immunotherapy. Adv Endocrinol Metab 1: 35–94Google Scholar
  41. 41.
    Kagi D, Vignaux F, Ledermann B, et al. (1994) Fas and perform pathways as major mechanisms of T cell-mediated cytotoxicity. Science 265: 528–530Google Scholar
  42. 42.
    Itoh N, Tsujimoto Y, Nagata S (1993) Effect of bcl-2 on Fas antigen-mediated cell death. J Immunol 151: 621–627Google Scholar
  43. 43.
    Juntti-Berggren L, Larsson O, Rorsman P, et al. (1993) Increased activity of L-type Ca2+ channels exposed to serum from patients with type 1 diabetes. Science 261: 86–90Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • H. Iwahashi
    • 1
  • T. Hanafusa
    • 1
  • Y. Eguchi
    • 2
  • H. Nakajima
    • 1
  • J. Miyagawa
    • 1
  • N. Itoh
    • 1
  • K. Tomita
    • 1
  • M. Namba
    • 1
  • M. Kuwajima
    • 1
    • 4
  • T. Noguchi
    • 3
    • 5
  • Y. Tsujimoto
    • 2
  • Y. Matsuzawa
    • 1
  1. 1.Second Department of Internal MedicineOsaka University Medical SchoolOsakaJapan
  2. 2.Department of Medical Genetics, Biomedical Research CenterOsaka University Medical SchoolOsakaJapan
  3. 3.Department of Nutrition and Physiological ChemistryOsaka University Medical SchoolOsakaJapan
  4. 4.Department of Laboratory Medicine, School of MedicineUniversity of TokushimaTokushimaJapan
  5. 5.Department of BiochemistryFukui Medical SchoolFukuiJapan

Personalised recommendations