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(Investigative Ophthalmology and Visual Science. 2000;41:2613-2622.)
© 2000 by The Association for Research in Vision and Ophthalmology, Inc.

IFN-{gamma} and LPS-Mediated IL-10–Dependent Suppression of Retinal Microglial Activation

Cathryn Broderick1, Linda Duncan1, Neil Taylor1 and Andrew D. Dick1,2

1 From the Department of Ophthalmology, University of Aberdeen, and 2 Division of Ophthalmology, University of Bristol, United Kingdom.

PURPOSE. Human retinal microglia (MG) express constitutively major histocompatability complex (MHC) class II molecules and have thus been highlighted as potential immunocompetent antigen-presenting cells (APCs). This study was undertaken to characterize microglial expression of coaccessory molecules and the functional changes in antigen expression, cytokine production, migration, and phagocytosis after proinflammatory stimulation.

METHODS. Fresh donor retinal MG were obtained and isolated using a percoll density gradient technique. Phenotypic characteristics used for isolating rodent microglia were applied and modified. Coaccessory molecule expression and intracellular cytokine production were assessed using three-color flow cytometric analysis in both freshly isolated and interferon (IFN){gamma}-lipopolysaccharide (LPS)–stimulated MG. Using five-millimeter retinal explants in culture, microglial migratory behavior, changes in cell surface antigen expression and phagocytic activity were documented.

RESULTS. MG could be clearly defined by the flow cytometric phenotype CD45lowCD11b+MHC class II+CD86lowCD40low. Freshly isolated MG showed mannose receptor–mediated uptake of dextran-FITC. MG migrated from explants, were adherent, and upregulated MHC class II expression. After IFN{gamma}-LPS stimulation of single-cell suspension of MG isolates, MHC class II expression was reduced, with an increase occurring in MG intracellular interleukin (IL)-10 and IL-10 production. Microglial migration from explants was reduced after IFN{gamma}-LPS stimulation.

CONCLUSIONS. These results highlight both phenotypic and behavioral characteristics that support an antigen-processing and -presenting capability of freshly isolated MG. However, proinflammatory stimulation with IFN{gamma}-LPS induces an IL-10–mediated downregulation of cell surface antigen expression and loss of migratory and phagocytic activity. Therefore, although equipped to act as APCs, MG are able to rapidly modulate their own function and behavior and as a result may have the potential to limit inflammation.




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