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S. P. Padmanabhan, R. N. Van Gelder; Identification of Autoantibodies in Sympathetic Ophthalmia by Serological Proteome Analysis. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3780.
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© ARVO (1962-2015); The Authors (2016-present)
To utilize a novel proteomics-based approach to characterize and compare the ocular autoantibody repertoires of healthy patients and those with sympathetic ophthalmia, and to identify candidate autoantigens that may be involved in the pathogenesis of sympathetic ophthalmia.
Four eligible subjects with sympathetic ophthalmia and four age- and sex-matched controls were enrolled. Total retinal, choroidal, ciliary body, and iris protein were extracted from human donor eyes (Mid-America Transplant, St. Louis, MO) and underwent two-dimensional electrophoretic separation. Candidate antigenic proteins were subsequently identified via Western analysis using participant serum as the source of primary antibody. Identical gels run in parallel provided material for antigen identification via mass spectrometry.
Enolase-1, heat shock protein 60, and vimentin in all four tissues were found to be preferentially antigenic in sympathetic ophthalmia subjects versus controls, which confirm the results of previous studies. Alpha- and beta- tubulin, shown to be antigenic in previous uveitis studies, produced an immune response that was limited to the retina. Other proteins identified as autoantigens across multiple tissues in the sympathetic ophthlamia subjects include carbonic anhydrase and annexin V, neither of which have been identified as possible candidate autoantigens in sympathetic ophthalmia. Also noted was the absence of antibody response to retinal S-antigen.
We have successfully demonstrated the use of serological proteome analysis as a means to identify human autoantibodies against ocular antigens in sympathetic ophthalmia. Our results confirm the results of previous studies and present several novel candidate autoantigens in sympathetic ophthalmia including carbonic anhydrase and annexin V.
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