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.

Key words

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

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