May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Immunoglobulins Produced by Human Retinal Pigment Epithelial Cells (RPE) Can Recognize Bruch's Membrane Proteins
Author Affiliations & Notes
  • L. Geng
    Department of Ophthalmology and Visual Science, University of Louisville, Kentucky Lions Eye Center, Louisville, Kentucky
  • S. Schaal
    Department of Ophthalmology and Visual Science, University of Louisville, Kentucky Lions Eye Center, Louisville, Kentucky
  • E. Bodek
    Department of Ophthalmology and Visual Science, University of Louisville, Kentucky Lions Eye Center, Louisville, Kentucky
  • H. J. Kaplan
    Department of Ophthalmology and Visual Science, University of Louisville, Kentucky Lions Eye Center, Louisville, Kentucky
  • T. H. Tezel
    Department of Ophthalmology and Visual Science, University of Louisville, Kentucky Lions Eye Center, Louisville, Kentucky
  • Footnotes
    Commercial Relationships L. Geng, None; S. Schaal, None; E. Bodek, None; H.J. Kaplan, None; T.H. Tezel, None.
  • Footnotes
    Support Supported in part by Career Development Award (T.H.T.) from Research to Prevent Blindness, Inc, NYC, NY.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2538. doi:
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    • Get Citation

      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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose:: To characterize the spatial and temporal deposition patterns of immunoglobulin within the human Bruch’s membrane and to identify their recognized epitopes.

Methods:: 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.

Results:: 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.

Conclusions:: Expression and/or deposition of immunoglobulins against Bruch’s membrane and RPE support the involvement of immune mechanism in cellular damage.

Keywords: Bruch's membrane • retinal pigment epithelium • proteomics 
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