RESUMEN
The cause of type 1 diabetes remains unknown. To dissect the link between hyperexpression of human leukocyte antigen (HLA) class I on the islet cells, we examined its expression in subjects with recent-onset type 1 diabetes. IHC showed seemingly pronounced hyperexpression in subjects with recent-onset type 1 diabetes, as well as in some nondiabetic subjects. In all subjects, HLA class I expression on exocrine tissue was low. However, no difference in the level of HLA class I expression was found between islet and exocrine tissue using Western blot, flow cytometry, real-time quantitative PCR, or RNA sequencing analyses. Also, the level of HLA class I expression on the messenger level was not increased in islets from subjects with recent-onset type 1 diabetes compared with that in nondiabetic subjects. Consistently, the HLA class I specific enhanceosome (NLRC5) and related transcription factors, as well as interferons, were not enhanced in islets from recent-onset type 1 diabetic subjects. In conclusion, a discrepancy in HLA class I expression in islets assessed by IHC was observed compared with that using quantitative techniques showing similar expression of HLA class I in islets and exocrine tissue in subjects with recent-onset type 1 diabetes, nor could any differences be found between type 1 diabetic and nondiabetic subjects. Results presented provide important clues for a better understanding on how this complex disease develops.
Asunto(s)
Diabetes Mellitus Tipo 1/metabolismo , Genes MHC Clase I , Islotes Pancreáticos/metabolismo , Páncreas Exocrino/metabolismo , Adolescente , Adulto , Anciano , Niño , Diabetes Mellitus Tipo 1/diagnóstico , Humanos , Interferones/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Persona de Mediana Edad , Factores de Transcripción/metabolismo , Transcriptoma , Adulto JovenAsunto(s)
Anticuerpos Monoclonales/metabolismo , Proteínas de la Cápside/inmunología , Diabetes Mellitus Tipo 1/metabolismo , Infecciones por Enterovirus/metabolismo , Enterovirus/metabolismo , Inmunohistoquímica/métodos , Oligopéptidos/inmunología , Páncreas/metabolismo , Femenino , Humanos , MasculinoRESUMEN
PURPOSE OF REVIEW: Type 1 diabetes (T1D) research is at present in a critical period of development and during the past few years several large phase III studies targeting T-cell autoimmunity in recent-onset patients with T1D failed to reach the primary endpoint. RECENT FINDINGS: Cause and pathogenesis of T1D remain largely unknown. In humans, insulitis is discrete, affects few islets and is present only in about one-third of patients with recent-onset T1D. The rapid increase in incidence of T1D argues against a decisive role for genetic factors and instead for the hypothesis that infectious agents, possibly entering the pancreas via the ductal compartment, are involved in disease pathogenesis. Repeated episodes of bacteria or virus-induced innate inflammations affecting only certain lobes of the pancreas fit well with the reported heterogeneity of the disease within the pancreas as well as with the slow progression over many years. SUMMARY: In humans there is limited support for T1D being primarily an autoimmune disease; instead available findings support the view that T1D can be regarded as an innate inflammatory disease affecting the entire pancreas, but with its main clinical manifestations emanating from the loss of the insulin-producing cells.
Asunto(s)
Autoanticuerpos/metabolismo , Autoinmunidad , Diabetes Mellitus Tipo 1/inmunología , Linfocitos T/metabolismo , Animales , Autoanticuerpos/inmunología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Humanos , Inmunoterapia/métodos , Inflamación/inmunología , Páncreas Exocrino/inmunología , Páncreas Exocrino/patología , Linfocitos T/inmunologíaRESUMEN
Current or recent enteroviral infections show an association with type 1 diabetes. However, evidence for this has mainly been generated using a particular mouse monoclonal antibody (clone 5-D8/1) which binds the viral capsid protein VP1. Difficulty in confirming these findings using other independent methods has led to the concern that this might be artefactual. To address this, we examined the potential cross-reactivity of clone 5-D8/1 with normal islet proteins. Western blotting, two-dimensional gel electrophoresis, and mass spectrometry were used to identify human islet proteins bound by the clone 5-D8/1. We found a distinct reactivity with two mitochondrial proteins, creatine kinase B-type and ATP synthase beta subunit. Immunohistochemistry using the clone 5-D8/1 revealed a granular cytoplasmic staining pattern in mitochondria-rich cells, ie hepatocytes, ductal epithelial cells, vascular endothelial cells, skeletal muscle cells, and the neoplastic salivary gland oncocytoma cells, whereas connective tissue and infiltrating immune cells were negative. Staining on islets of Langerhans from subjects with recent-onset type 1 diabetes, but not on isolated human islets infected in vitro with enteroviruses, could be blocked after mixing the clone 5-D8/1 with the mitochondrial proteins. Collectively, our data show that the clone 5-D8/1 detects two human mitochondrial enzymes in addition to enteroviral VP1. The notion that the previously reported VP1 positivity in islets of recent-onset type 1 diabetes patients could reflect cross-reactivity to native islet proteins and not the presence of EV is supported by difficulties in demonstrating EV infection by independent techniques such as PCR or in situ hybridization. These findings call for revisiting the presence of enteroviruses in pancreatic islets of patients with type 1 diabetes.
Asunto(s)
Anticuerpos/inmunología , Proteínas de la Cápside/inmunología , Creatina Quinasa/inmunología , Diabetes Mellitus Tipo 1/inmunología , Infecciones por Enterovirus/inmunología , Enterovirus/inmunología , Islotes Pancreáticos/inmunología , ATPasas de Translocación de Protón Mitocondriales/inmunología , Artefactos , Western Blotting , Reacciones Cruzadas , Diabetes Mellitus Tipo 1/enzimología , Diabetes Mellitus Tipo 1/virología , Electroforesis en Gel Bidimensional , Infecciones por Enterovirus/complicaciones , Infecciones por Enterovirus/virología , Humanos , Inmunohistoquímica , Islotes Pancreáticos/enzimología , Islotes Pancreáticos/virología , Proteómica/métodos , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem , Técnicas de Cultivo de TejidosRESUMEN
The cause of type 1 diabetes (T1D) remains unknown; however, a decisive role for environmental factors is recognized. The increased incidence of T1D during the last decades, as well as regional differences, is paralleled by differences in the intestinal bacterial flora. A new animal model was established to test the hypothesis that bacteria entering the pancreatic ductal system could trigger ß-cell destruction and to provide new insights to the immunopathology of the disease. Obtained findings were compared with those present in two patients dying at onset of T1D. Different bacterial species, present in the human duodenum, instilled into the ductal system of the pancreas in healthy rats rapidly induced cellular infiltration, consisting of mainly neutrophil polymorphonuclear cells and monocytes/macrophages, centered around the pancreatic ducts. Also, the islets of Langerhans attracted polymorphonuclear cells, possibly via release of IL-6, IL-8, and monocyte chemotactic protein 1. Small bleedings or large dilatations of the capillaries were frequently found within the islets, and several ß-cells had severe hydropic degeneration (ie, swollen cytoplasm) but with preserved nuclei. A novel rat model for the initial events in T1D is presented, revealing marked similarities with the morphologic findings obtained in patients dying at onset of T1D and signifying a decisive role for bacteria in eliciting an adverse innate immunity response. The present findings support the hypothesis that T1D is an organ-specific inflammatory disease.