title: Mitochondrial DNA Activates the NLRP3 Inflammasome and Predisposes to Type 1 Diabetes in Murine Model.
abstract: Although a correlation between polymorphisms of NOD-like receptor family-pyrin domain containing 3 (NLRP3) and predisposition to type 1 diabetes (T1D) has been identified, the potential function and activation of the NLRP3 inflammasome in T1D have not been clarified. The present study shows that non-obese diabetic mice exhibited increased NLRP3, and pro-IL-1β gene expression in pancreatic lymph nodes (PLNs). Similar increases in gene expression of NLRP3, apoptosis associated speck like protein (ASC) and pro-IL-1β were induced by multiple low doses of streptozotocin (STZ) in C57BL/6 mice. In addition, diabetic C57BL/6 mice also exhibited increased IL-1β protein expression in the pancreatic tissue at day 7, which remained elevated until day 15. Diabetic mice also showed increased positive caspase-1 macrophages in the PLNs, which were decreased in NLRP3-/-mice, but not in ASC-/-mice, after STZ treatment. NLRP3- and IL-1R-deficient mice, but not ASC-deficient mice, showed reduced incidence of diabetes, less insulitis, lower hyperglycemia, and normal insulin levels compared to wild-type (WT) diabetic mice. Notably, these mice also displayed a decrease in IL-17-producing CD4 and CD8 T cells (Th17 and Tc17) and IFN-γ-producing CD4 and CD8 T cells (Th1 and Tc1) in the PLNs. Following STZ treatment to induce T1D, NLRP3-deficient mice also exhibited an increase in myeloid-derived suppressor cell and mast cell numbers in the PLNs along with a significant increase in IL-6, IL-10, and IL-4 expression in the pancreatic tissue. Interestingly, diabetic mice revealed increased circulating expression of genes related to mitochondrial DNA, such as cytochromeband cytochromec, but not NADH dehydrogenase subunit 6 (NADH). Mitochondrial DNA (mDNA) from diabetic mice, but not from non-diabetic mice, induced significant IL-1β production and caspase-1 activation by WT macrophages, which was reduced in NLRP3-/-macrophages. Finally, mDNA administrationin vivoincreased Th17/Tc17/Th1/Tc1 cells in the PLNs and precipitated T1D onset, which was abolished in NLRP3-/-mice. Overall, our results demonstrate that mDNA-mediated NLRP3 activation triggers caspase-1-dependent IL-1β production and contributes to pathogenic cellular responses during the development of STZ-induced T1D.
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Daniela
authors.initials: D
authors.lastname: Carlos
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Frederico R C
authors.initials: FR
authors.lastname: Costa
authors.affiliation: Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Camila A
authors.initials: CA
authors.lastname: Pereira
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Fernanda A
authors.initials: FA
authors.lastname: Rocha
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Juliana N U
authors.initials: JN
authors.lastname: Yaochite
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Gabriela G
authors.initials: GG
authors.lastname: Oliveira
authors.affiliation: Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Fernando S
authors.initials: FS
authors.lastname: Carneiro
authors.affiliation: Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Rita C
authors.initials: RC
authors.lastname: Tostes
authors.affiliation: Department of Pathology and Legal Medicine, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Simone G
authors.initials: SG
authors.lastname: Ramos
authors.affiliation: Department of Molecular and Cell Biology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: Dario S
authors.initials: DS
authors.lastname: Zamboni
authors.affiliation: Department of Immunology, Institute of Biomedical Sciences (ICB), University of São Paulo , São Paulo , Brazil.
authors.forename: Niels O S
authors.initials: NO
authors.lastname: Camara
authors.affiliation: University of Orleans and CNRS, INEM, Molecular Immunology, UMR6218, Orleans, France; IDM, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Ryffel
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo , Ribeirao Preto , Brazil.
authors.forename: João S
authors.initials: JS
authors.lastname: Silva
keywords: IFNγ-producing CD4 T cells (Th1); IFNγ-producing CD8 T cells (Tc1); IL-17-producing CD4 T cells (Th17); IL-17-producing CD8 T cells (Tc17); NLRP3 inflammasome; cytokines; type 1 diabetes