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L. Geng, S. Schaal, E. Bodek, H. J. Kaplan, T. H. Tezel; Immunoglobulins Produced by Human Retinal Pigment Epithelial Cells (RPE) Can Recognize Bruch's Membrane Proteins. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2538.
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To characterize the spatial and temporal deposition patterns of immunoglobulin within the human Bruch’s membrane and to identify their recognized epitopes.
Topographical and age-related changes of immunoglobulin expression of RPE and its deposition within Bruch’s membrane were studied in young (<50) and old (>65) donors. For this purpose human RPE and Bruch’s membrane proteins were extracted from 4 donor eyes (ages: 41-66) and focused using non-liner (pH 3-10) 7.7cm IEF strips. Proteins were further separated in the second dimension on 4-12% Bis-Tris gels. Proteins recognized by RPE and Bruch’s membrane immunoglobulins were determined by 2-D immunoblotting where RPE and Bruch’s membrane proteins from the same donor were used as a primary antibody. Recognized proteins were identified using mass spectrometry. Results were confirmed with Western blotting and RT-PCR.
Immunoglobulins are major components of human RPE and Bruch’s membrane proteome, but not photoreceptors. Immunoglobulins expressed by RPE cells can recognize 29 extracellular matrix proteins within Bruch’s membrane. Similarly, increased immunoglobulins within the Bruch’s membrane of elderly donors recognize 32 RPE proteins.
Expression and/or deposition of immunoglobulins against Bruch’s membrane and RPE support the involvement of immune mechanism in cellular damage.
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