Blood Glucose Levels Regulate Pancreatic β-Cell Proliferation during Experimentally-Induced and Spontaneous Autoimmune Diabetes in Mice

Klaus Pechhold,Greg Walker,Kerstin Koczwara,Xiaolong Zhu,David M Harlan,Victor S Harrison,Janet Lee,Kathrin Maedler

doi:10.1371/journal.pone.0004827

Abstract

BackgroundType 1 diabetes mellitus is caused by immune-mediated destruction of pancreatic β-cells leading to insulin deficiency, impaired intermediary metabolism, and elevated blood glucose concentrations. While at autoimmune diabetes onset a limited number of β-cells persist, the cells' regenerative potential and its regulation have remained largely unexplored. Using two mouse autoimmune diabetes models, this study examined the proliferation of pancreatic islet ß-cells and other endocrine and non-endocrine subsets, and the factors regulating that proliferation.Methodology and Principal FindingsWe adapted multi-parameter flow cytometry techniques (including DNA-content measurements and 5′-bromo-2′-deoxyuridine [BrdU] incorporation) to study pancreatic islet single cell suspensions. These studies demonstrate that β-cell proliferation rapidly increases at diabetes onset, and that this proliferation is closely correlated with the diabetic animals' elevated blood glucose levels. For instance, we show that when normoglycemia is restored by exogenous insulin or islet transplantation, the β-cell proliferation rate returns towards low levels found in control animals, yet surges when hyperglycemia recurs. In contrast, other-than-ß endocrine islet cells did not exhibit the same glucose-dependent proliferative responses. Rather, disease-associated alterations of BrdU-incorporation rates of δ-cells (minor decrease), and non-endocrine islet cells (slight increase) were not affected by blood glucose levels, or were inversely related to glycemia control after diabetes onset (α-cells).ConclusionWe conclude that murine β-cells' ability to proliferate in response to metabolic need (i.e. rising blood glucose concentrations) is remarkably well preserved during severe, chronic β-cell autoimmunity. These data suggest that timely control of the destructive immune response after disease manifestation could allow spontaneous regeneration of sufficient β-cell mass to restore normal glucose homeostasis.

Highlights

Type 1 Diabetes Mellitus (T1DM) is an autoimmune disease characterized by insulin deficiency and the loss of glycemia control, and is caused by the T-cell mediated destruction of pancreatic insulin-producing ß cells
Flow cytometry demonstrates that autoimmune diabetes stimulates ß-cell cycle progression
Reasoning that adapting flow cytometry-based techniques to pancreas research would greatly facilitate a more quantitative analysis of b-cell proliferation in islets undergoing autoimmune destruction, we have developed protocols to isolate islets from naıve and diabetic mice, dissociate them into single cells, and identify their endocrine cell lineage by co-staining for intracytoplasmic insulin, glucagon, and somatostatin

Summary

Introduction

Type 1 Diabetes Mellitus (T1DM) is an autoimmune disease characterized by insulin deficiency and the loss of glycemia control, and is caused by the T-cell mediated destruction of pancreatic insulin-producing ß cells. It becomes clinically apparent rather late during its chronic progressive course, and has long been considered irreversible due to the assumption that b-cells have no or only limited regenerative capacity and are unable to withstand the continuing ß-cell autoimmunity. Recent reports have suggested that adult b-cells might be able to expand under certain conditions [1], and experimental rodent models have confirmed this notion in general [2]. Using two mouse autoimmune diabetes models, this study examined the proliferation of pancreatic islet ß-cells and other endocrine and non-endocrine subsets, and the factors regulating that proliferation

Methods

Results

Conclusion