Cytokines released by islet infiltrating immune cells play a crucial role in ß-cell dysfunction and apoptotic cell death in the pathogenesis of type 1 diabetes. RNA studies revealed complex pathways of genes being activated or suppressed during this ß-cell attack. The aim of the present study was to analyze protein changes in insulin-producing INS-1E cells exposed to inflammatory cytokines in vitro using 2D-DIGE. Within two different pH ranges we observed 2214 +/-164 (pH 4-7) and 1641 +/-73 (pH 6-9) spots. Analysis at 3 different time points (1, 4 and 24 hours of cytokine exposure) revealed that the major changes were taking place only after 24 hours. At this time point 158 proteins were altered in expression (4.1%, n=4, p=0.01) by a combination of IL-1b and IFN-g, whereas only 42 and 23 proteins were altered by either of the cytokines alone, giving rise to 199 distinct differential expressed spots. Identification of 141 of these by MALDI-TOF/TOF, revealed proteins playing a role in insulin secretion, cytoskeleton organization, protein and RNA metabolism, as well as proteins associated with endoplasmic reticulum- and oxidative stress / defense. We investigated the interactions of these proteins and discovered a significant interaction network (p<1.27e-05) containing 42 of the identified proteins. This network analysis suggests that proteins of different pathways act coordinately in a ß-cell dysfunction / apoptotic ß-cell death - interactome. In addition the data suggest a central role for chaperones and proteins playing a role in RNA metabolism. As many of these identified proteins are regulated at the protein level or undergo post-translational modifications, a proteomic approach, as performed in this study, is required to provide adequate insight into the mechanisms leading to ß-cell dysfunction and apoptosis. The present findings may open new avenues for the understanding and prevention of ß-cell loss in type 1 diabetes.