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Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden; Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium
Address all correspondence and requests for reprints to: D. L. Eizirik, Diabetes Research Center, Vrije Universiteit Brussel, Laarbeeklaan 103 B-1090 Brussels, Belgium. E-mail: deizirik{at}mebo.vub.ac.be
We have previously observed that a 6-day exposure of human pancreatic
islets to a combination of cytokines (interleukin-1ß 50 U/ml + tumour
necrosis factor-
1000 U/ml + interferon-
1000 U/ml) severely
impairs ß-cell functions. In the present study, we examined whether
this condition affects DNA integrity and viability of human islet
cells. Cells were studied after 3, 6, and 9 days of cytokine treatment
by both single cell gel electrophoresis (the "comet assay," a
sensitive method for detection of DNA strand breaks) and by a
cytotoxicity assay using the DNA binding dyes Hoechst 33342 and
propidium iodide as indices for the number of viable, necrotic, and
apoptotic cells. Cytokine treatment for 6 and 9 days resulted in a 50%
increase in comet length (P < 0.01
vs. controls), indicating DNA strand breaks, as well as
in a significant increase in the number of apoptotic cells
(P < 0.02 vs. controls), but not in
the number of necrotic cells. The arginine analogs
NG-nitro-L-arginine and
NG-monomethyl-L-arginine prevented nitric oxide
formation by the cytokines but did not interfere with cytokine-induced
DNA strand breaks and apoptosis. The present data suggest that
prolonged (6–9 days) exposure of human pancreatic islets to a mixture
of cytokines induces DNA strand breaks and cell death by apoptosis.
These deleterious effects of cytokines appear to be independent of
nitric oxide generation.
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H. Heimberg, Y. Heremans, C. Jobin, R. Leemans, A. K. Cardozo, M. Darville, and D. L. Eizirik Inhibition of Cytokine-Induced NF-{kappa}B Activation by Adenovirus-Mediated Expression of a NF-{kappa}B Super-Repressor Prevents {beta}-Cell Apoptosis Diabetes, October 1, 2001; 50(10): 2219 - 2224. [Abstract] [Full Text]
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A. E. Karlsen, S. G. Ronn, K. Lindberg, J. Johannesen, E. D. Galsgaard, F. Pociot, J. H. Nielsen, T. Mandrup-Poulsen, J. Nerup, and N. Billestrup Suppressor of cytokine signaling 3 (SOCS-3) protects beta -cells against interleukin-1beta - and interferon-gamma -mediated toxicity PNAS, September 26, 2001; (2001) 211445998. [Abstract] [Full Text] [PDF]
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M. I. Darville and D. L. Eizirik Cytokine Induction of Fas Gene Expression in Insulin-Producing Cells Requires the Transcription Factors NF-{kappa}B and C/EBP Diabetes, August 1, 2001; 50(8): 1741 - 1748. [Abstract] [Full Text] [PDF]
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D. Liu, M. Darville, and D. L. Eizirik Double-Stranded Ribonucleic Acid (RNA) Induces {beta}-Cell Fas Messenger RNA Expression and Increases Cytokine-Induced {beta}-Cell Apoptosis Endocrinology, June 1, 2001; 142(6): 2593 - 2599. [Abstract] [Full Text] [PDF]
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