Inhibition of Cytokine-Induced NF-κB Activation by Adenovirus-Mediated Expression of a NF-κB Super-Repressor Prevents β-Cell Apoptosis

Inhibition of Cytokine-Induced NF-κB Activation by Adenovirus-Mediated Expression of a NF-κB Super-Repressor Prevents β-Cell Apoptosis Harry Heimberg 1 , Yves Heremans 1 , Christian Jobin 2 , Ruth Leemans 1 , Alessandra K. Cardozo 1 , Martine Darville 1 and Décio L. Eizirik 1 1 Diabetes Research Cen...

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Veröffentlicht in:Diabetes (New York, N.Y.) N.Y.), 2001-10, Vol.50 (10), p.2219-2224
Hauptverfasser: Heimberg, Harry, Heremans, Yves, Jobin, Christian, Leemans, Ruth, Cardozo, Alessandra K., Darville, Martine, Eizirik, Décio L.
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Sprache:eng
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Zusammenfassung:Inhibition of Cytokine-Induced NF-κB Activation by Adenovirus-Mediated Expression of a NF-κB Super-Repressor Prevents β-Cell Apoptosis Harry Heimberg 1 , Yves Heremans 1 , Christian Jobin 2 , Ruth Leemans 1 , Alessandra K. Cardozo 1 , Martine Darville 1 and Décio L. Eizirik 1 1 Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium 2 Division of Digestive Diseases and Nutrition, University of North Carolina, Chapel Hill, North Carolina Abstract Cytokine-induced β-cell death is an important event in the pathogenesis of type 1 diabetes. The transcription factor nuclear factor-κB (NF-κB) is activated by interleukin-1β (IL-1β), and its activity promotes the expression of several β-cell genes, including pro- and anti-apoptotic genes. To elucidate the role of cytokine (IL-1β + γ-interferon [IFN-γ])-induced expression of NF-κB in β-cell apoptosis, rat β-cells were infected with the recombinant adenovirus AdIκB (SA)2 , which contained a nondegradable mutant form of inhibitory κB (IκB (SA)2 , with S32A and S36A) that locks NF-κB in a cytosolic protein complex, preventing its nuclear action. Expression of IκB (SA)2 inhibited cytokine-stimulated nuclear translocation and DNA-binding of NF-κB. Cytokine-induced gene expression of several NF-κB targets, namely inducible nitric oxide synthase, Fas, and manganese superoxide dismutase, was prevented by AdIκB (SA)2 , as established by reverse transcriptase–polymerase chain reaction, protein blot, and measurement of nitrite in the medium. Finally, β-cell survival after IL-1β + IFN-γ treatment was significantly improved by IκB (SA)2 expression, mostly through inhibition of the apoptotic pathway. Based on these findings, we conclude that NF-κB activation, under in vitro conditions, has primarily a pro-apoptotic function in β-cells. Footnotes Address correspondence and reprint requests to H. Heimberg, Diabetes Research Center, Vrije Universiteit Brussel, Laarbeeklaan 103, B-1090 Brussels, Belgium. E-mail: hheimber{at}vub.vub.ac.be . H.H. and Y.H. contributed equally to this work. Received for publication 6 October 2000 and accepted in revised form 13 July 2001. ANOVA, analysis of variance; FasL, ligand protein for Fas; GAPDH, glyceraldehyde-phosphate dehydrogenase; GFP, green fluorescent protein; IFN-γ, γ-interferon IκB, inhibitory κB; IL-1β, interleukin-1β; iNOS, inducible nitric oxide synthase; MnSOD, manganese superoxide dismutase; MOI, multiplicity of infection; NO, nitric oxide; PCR, polymerase chain react
ISSN:0012-1797
1939-327X
DOI:10.2337/diabetes.50.10.2219