Abstract
Abstract: :
Purpose:To investigate macrophage expression of tissue factor (TF) and retinal pigment epithelium (RPE) expression of monocyte colonization protein (MCP) in surgically-excised choroidal neovascular membranes.Methods:Five surgically-excised choroidal neovascular (CNV) membranes from patients with ocular histoplasmosis syndrome (OHS) and age-related macular degeneration (AMD) were evaluated by confocal scanning laser microscopy. The CNV specimens were deparaffinized, blocked with horse serum washed with PBS, and incubated with CD68 IgG or MCP (1:100) at 4°C. The tissue was washed, incubated in donkey anti-mouse IgG-Rhodamine Red conjugate (1:200), washed and incubated in goat anti-mouse Fab fragments (1:50) for two hours. The tissue was washed, incubated overnight with TF or CK18 (10µg/ml), washed, incubated with donkey anti-mouse IgG-FITC (1:200) conjugate for 2°, washed and incubated in clearing solution, and coverslipped. A Biorad confocal scanning laser microscope was used to obtain digitized images.Results:All specimens contained RPE that exhibited dual labeling for CK18 and MCP. All specimens contained macrophages exhibiting dual labeling for CD68 and TF.Conclusion:These results indicate CNV from both OHS and AMD contain RPE that expresses MCP and macrophages that express TF. The RPE may attract macrophages from the systemic circulation and the macrophages express TF, which acts as a scaffold for the CNV.
Keywords: 308 age-related macular degeneration • 346 choroid: neovascularization • 471 microscopy: confocal/tunneling