Type 1 Diabetes (T1D) is considered to be a T-helper- (Th-) 1 autoimmune disease; however, T1D pathogenesis likely involves many factors, and sufficient tools for autoreactive T cell detection for the study of this disease are currently lacking. In this study, using gene expression microarrays, we analysed the effect of diabetes-associated autoantigens on peripheral blood mononuclear cells (PBMCs) with the purpose of identifying (pre)diabetes-associated cell processes. Twelve patients with recent onset T1D, 18 first-degree relatives of the TD1 patients (DRL; 9/18 autoantibody positive), and 13 healthy controls (DV) were tested. PBMCs from these individuals were stimulated with a cocktail of diabetes-associated autoantigens (proinsulin, IA-2, and GAD65-derived peptides). After 72 hours, gene expression was evaluated by high-density gene microarray. The greatest number of functional differences was observed between relatives and controls (69 pathways), from which 15% of the pathways belonged to “immune response-related” processes. In the T1D versus controls comparison, more pathways (24%) were classified as “immune response-related.” Important pathways that were identified using data from the T1D versus controls comparison were pathways involving antigen presentation by MHCII, the activation of Th17 and Th22 responses, and cytoskeleton rearrangement-related processes. Genes involved in Th17 and TGF-beta cascades may represent novel, promising (pre)diabetes biomarkers. 1. Introduction Type 1 Diabetes (T1D) is considered to be a T-helper- (Th-) 1 autoimmune disease and is characterised by a lack of insulin, which is caused by the autoimmune destruction of insulin-producing pancreatic beta cells [1, 2]. Th1 lymphocytes are responsible for the infiltration of the islets of Langerhans and for the cytokine release that facilitates the destruction of beta cells by cytotoxic (Tc) lymphocytes. Due to this progressive damage, there is either insufficient or no production of insulin, leading to the first clinical signs of T1D. At the first appearance of clinical symptoms, most notably those associated with hyperglycaemia, nearly 80% of the beta cells have been destroyed, rendering the individual dependent on insulin injections [2, 3]. In patients presenting with recent T1D onset, there are various interventions that may stop, or at least delay, pancreatic beta cell destruction; however, these therapies are unable to reverse the patient’s lifelong dependency on insulin injections because beta cell proliferation and their capacity for regeneration are limited. To save
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