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Cell Death Dis ; 7(9): e2367, 2016 09 08.
Article in English | MEDLINE | ID: mdl-27607582

ABSTRACT

Retinal pigment epithelial (RPE) cells can undergo different forms of cell death, including autophagy-associated cell death during age-related macular degeneration (AMD). Failure of macrophages or dendritic cells (DCs) to engulf the different dying cells in the retina may result in the accumulation of debris and progression of AMD. ARPE-19 and primary human RPE cells undergo autophagy-associated cell death upon serum depletion and oxidative stress induced by hydrogen peroxide (H2O2). Autophagy was revealed by elevated light-chain-3 II (LC3-II) expression and electron microscopy, while autophagic flux was confirmed by blocking the autophago-lysosomal fusion using chloroquine (CQ) in these cells. The autophagy-associated dying RPE cells were engulfed by human macrophages, DCs and living RPE cells in an increasing and time-dependent manner. Inhibition of autophagy by 3-methyladenine (3-MA) decreased the engulfment of the autophagy-associated dying cells by macrophages, whereas sorting out the GFP-LC3-positive/autophagic cell population or treatment by the glucocorticoid triamcinolone (TC) enhanced it. Increased amounts of IL-6 and IL-8 were released when autophagy-associated dying RPEs were engulfed by macrophages. Our data suggest that cells undergoing autophagy-associated cell death engage in clearance mechanisms guided by professional and non-professional phagocytes, which is accompanied by inflammation as part of an in vitro modeling of AMD pathogenesis.


Subject(s)
Autophagy/drug effects , Culture Media, Serum-Free/pharmacology , Hydrogen Peroxide/pharmacology , Macrophages/drug effects , Phagocytosis/drug effects , Retinal Pigment Epithelium/drug effects , Adenine/analogs & derivatives , Adenine/pharmacology , Autophagy/genetics , Biomarkers/metabolism , Cell Line , Chloroquine/pharmacology , Coculture Techniques , Gene Expression , Genes, Reporter , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Humans , Interleukin-6/biosynthesis , Interleukin-6/metabolism , Interleukin-8/biosynthesis , Interleukin-8/metabolism , Macrophages/cytology , Macrophages/immunology , Macular Degeneration/genetics , Macular Degeneration/immunology , Macular Degeneration/pathology , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/immunology , Models, Biological , Oxidative Stress , Primary Cell Culture , Retinal Pigment Epithelium/immunology , Retinal Pigment Epithelium/pathology , Triamcinolone/pharmacology
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