Wagner, Leslie E.Melnyk, OlhaTurner, AbigailDuffett, Bryce E.Muralidharan, CharanyaMartinez-Irizarry, Michelle M.Arvin, Matthew C.Orr, Kara S.Manduchi, ElisabettaKaestner, Klaus H.Brozinick, Joseph T.Linnemann, Amelia K.2025-04-172025-04-172025-02-20Wagner LE, Melnyk O, Turner A, et al. IFN-α Induces Heterogenous ROS Production in Human β-Cells. Preprint. bioRxiv. 2025;2025.02.19.639120. Published 2025 Feb 20. doi:10.1101/2025.02.19.639120https://hdl.handle.net/1805/47112Type 1 diabetes (T1D) is a multifactorial disease involving genetic and environmental factors, including viral infection. We investigated the impact of interferon alpha (IFN-α), a cytokine produced during the immune response to viral infection or the presence of un-edited endogenous double-stranded RNAs, on human β-cell physiology. Intravital microscopy on transplanted human islets using a β-cell-selective reactive oxygen species (ROS) biosensor (RIP1-GRX1-roGFP2), revealed a subset of human β-cells that acutely produce ROS in response to IFN-α. Comparison to Integrated Islet Distribution Program (IIDP) phenotypic data revealed that healthier donors had more ROS accumulating cells. In vitro IFN-α treatment of human islets similarly elicited a heterogenous increase in superoxide production that originated in the mitochondria. To determine the unique molecular signature predisposing cells to IFN-α stimulated ROS production, we flow sorted human islets treated with IFN-α. RNA sequencing identified genes involved in inflammatory and immune response in the ROS-producing cells. Comparison with single cell RNA-Seq datasets available through the Human Pancreas Analysis Program (HPAP) showed that genes upregulated in ROS-producing cells are enriched in control β-cells rather than T1D donors. Combined, these data suggest that IFN-α stimulates mitochondrial ROS production in healthy human β-cells, potentially predicting a more efficient antiviral response.en-USAttribution-NonCommercial-NoDerivatives 4.0 InternationalType 1 diabetes (T1D)Genetic factorsEnvironmental factorsViral infectionArticle