Abstract
AIMS/BACKGROUND: Endotoxin induced uveitis has been regarded as a model for acute anterior uveitis and until now little was known about choroidal involvement. The aim of this study was to investigate changes in macrophages and MHC class II positive cells in the choroid of Lewis rats during endotoxin induced uveitis. METHODS: Choroid-sclera wholemounts were isolated from normal Lewis rats and at different time points--4, 8, 16, 24, 48, 72, and 96 hours, and 7, 10, and 14 days after a footpad injection of 200 micrograms of lipopolysaccharide (LPS). Immunohistochemistry was performed using the monoclonal antibodies ED1 (monocytes, macrophages, dendritic cells), and OX6 (MHC class II antigen). RESULTS: In normal rats, two layers of macrophages were identified in the choroid; a layer located immediately beneath the retinal pigment epithelium (RPE) and a layer bordering the sclera. The density of ED1 positive cells in the layer bordering the RPE cells was 902 (SD 132) cells/mm2 whereas the scleral layer had a cell density of 389 (73) cells/mm2. Based on morphology, positive cells could be divided into two main categories; pleomorphic/round cells and dendritiform cells with varying appearances, with the latter being predominant in normal eyes. A network of MHC class II positive dendritic cells was found in the choroid, beneath the RPE, with a density of 659 (96) cells/mm2. No MHC class II positive cells were found in the macrophage layer bordering the sclera. LPS injection caused a massive influx of ED1 positive macrophages in the area below the RPE cells but did not result in an influx of macrophages at the scleral side of the choroid. The infiltrate reached a maximum at 16 hours following LPS injection and decreased at 96 hours. The morphology of the infiltrating cells was pleomorphic/round at early stages of inflammation and changed into a dendritiform cell population later. The number of MHC class II positive cells on the anterior side of the choroid increased 8 hours after injection and reached a peak at 72-96 hours. MHC class II positive cells were not observed in the vicinity of the sclera at any time after LPS injection. Both resident and MHC class II positive dendritic cell numbers returned to normal values at day 14 following LPS injection. CONCLUSIONS: These results indicate that the choroid is severely inflamed after systemic LPS administration to Lewis rats and suggests that endotoxin induced uveitis may serve as a model for generalised uveitis in humans.