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Barbara J. McLaughlin, Wei Fan, Jing Juan Zheng, Hui Cai, Lucian V. Del Priore, Nalini S. Bora, Henry J. Kaplan; Novel Role for a Complement Regulatory Protein (CD46) in Retinal Pigment Epithelial Adhesion. Invest. Ophthalmol. Vis. Sci. 2003;44(8):3669-3674. doi: 10.1167/iovs.02-0813.
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purpose. There is increasing evidence that the complement system may play a significant role in one of the leading diseases causing blindness in the elderly population, age-related macular degeneration. In this study, a novel role in the retina for a regulatory protein in the complement system, CD46, is proposed.
methods. The retinal pigment epithelium (RPE) was obtained from human donor eyes as well as human immortalized RPE cell lines (ARPE19). Immunohistochemistry and confocal microscopy were used to immunolocalize CD46 and β1 integrin. Immunoprecipitation experiments with antibodies to either CD46 or β1 integrin were performed on RPE cell lysates. A cell adhesion assay was used to determine the proportion of RPE cells that adhere to Bruch’s membrane explants from donor eyes.
results. Immunohistochemistry and confocal microscopy demonstrated that CD46 was polarized to the basal surface of the RPE along with β1 integrin, shown previously to be involved in RPE adhesion. Immunoprecipitation experiments demonstrated that CD46 and β1 integrin coprecipitated from RPE cell lysates when either protein was used as the precipitating antibody. The adhesion assay showed that antibodies to either CD46 or β1 integrin reduced RPE adhesion to the surface of Bruch’s membrane compared with the control.
conclusions. These findings suggest that this complement regulatory protein, which protects host cells from autologous complement attack, may have a functional interaction with β1 integrin in the eye that is related to RPE adhesion to its basement membrane and Bruch’s membrane.
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