Disease resistant, NOD-related strains reveal checkpoints of immunoregulation in the pancreas

Abstract

The autoimmune diabetic NOD mouse serves as a model for human type 1 diabetes. Disease development is due to islet β cell destruction in the context of immune cell infiltration of islets and inflammatory changes throughout the pancreas. In the present study we tried to identify immune reactivity patterns in the pancreas associated with diabetes resistance in NOD-related mouse strains. The pancreata of diabetes-prone female NOD/LtJ, NOD/Bom and of genetically related but diabetes-resistant strains; NOR, NON, NON.NOD-H2^g7, NOD.NON-H-2^nbl were obtained at the age of 70 days for semiquantitative analysis of insulitis and of mRNA expression by reverse transcriptase PCR. In addition, the response to a single dose of cyclophosphamide for synchronizing and accelerating the progression of insulitis was determined. The progression of insulitis and immune gene expression in response to cyclophosphamide revealed characteristic differences between the six strains. NOD/LtJ and NOD/Bom mice were found significantly to upregulate pancreatic IL-12p40 and IL-18 expression after cyclophosphamide treatment, followed by an increase in IFN-γ mRNA levels. In contrast, the two MHC-haplotype H-2^nbl expressing strains either upregulated neither IL-12/IL-18 nor IFN-γ gene expression. The two strains sharing MHC haplotype H-2^g7 expression with NOD did respond to cyclophosphamide with IL-12p40/IL-18 gene expression. However, NON.NOD-H-2^g7 mice failed to progress to IFN-γ gene expression. NOR mice progressed to IFN-γ expression but exhibited sustained IL-4 gene expression. Only severe intra-insulitis was associated with the expression of inducible NO synthase. The comparison of diabetes-prone and diabetes-resistant strains revealed three checkpoints of immune regulation in the pancreas. The earliest checkpoint is the induction of an IL-12p40/IL-18 response in innate immune or antigen-presenting cells. The next level of control is at the induction of IFN-γ gene expression, and a third checkpoint is the maintenance or loss of antagonistic Th2 type reactions.