Human enteroviral infection impairs autophagy in clonal INS(832/13) cells and human pancreatic islet cells.

Diabetologia
Anya WernerssonCorrado M Cilio

Abstract

Human enteroviral infections are suggested to be associated with type 1 diabetes. However, the mechanism by which enteroviruses can trigger disease remains unknown. The present study aims to investigate the impact of enterovirus on autophagy, a cellular process that regulates beta cell homeostasis, using the clonal beta cell line INS(832/13) and human islet cells as in vitro models. INS(832/13) cells and human islet cells were infected with a strain of echovirus 16 (E16), originally isolated from the stool of a child who developed type 1 diabetes-associated autoantibodies. Virus production and release was determined by 50% cell culture infectious dose (CCID50) assay and FACS analysis. The occurrence of autophagy, autophagosomes, lysosomes and autolysosomes was detected by western blot, baculoviral-mediated expression of microtubule-associated protein light chain 3 (LC3)II-GFP and LysoTracker Red, and quantified by Cellomics ArrayScan. Autophagy was also monitored with a Cyto-ID detection kit. Nutrient deprivation (low glucose [2.8 mmol/l]), amino acid starvation (Earle's Balanced Salt Solution [EBSS]) and autophagy-modifying agents (rapamycin and chloroquine) were used in control experiments. Insulin secretion and the expressio...Continue Reading

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Citations

Aug 3, 2021·American Journal of Physiology. Endocrinology and Metabolism·Charanya Muralidharan, Amelia K Linnemann

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Methods Mentioned

BETA
scraping
PCR
ELISA
ubiquitination

Software Mentioned

CytExpert
Prism
GraphPad Prism
Cellomics

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