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E. W. J. Dervan, H. Chen, S. Ho, N. Brummel, M. Haralambova, E. Gould, D. Wallace, J. H. M. Prehn, C. J. O'Brien, D. Murphy; Protein Macroarray Profiling of Serum Autoantibodies in Pseudoexfoliation Glaucoma. Invest. Ophthalmol. Vis. Sci. 2010;51(13):5220.
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Complex repertoires of IgG autoantibodies have been detected against ocular antigens in patients with glaucoma. Our aim was to identify and characterize the IgG autoantibody repertoires in sera of patients with pseudoexfoliation glaucoma (PXFG) using protein macroarrays.
Serum samples of 21 patients with PXFG and 19 age and sex matched controls were profiled on high-density colony protein macroarrays expressing His-tagged recombinant human proteins derived from a human fetal brain cDNA library. Statistically prevalent expression clones in the PXFG group were sequenced. mRNA expression of identified antigens was examined in the rat ganglion cell line (RGC-5) and human brain and optic nerve cDNA. The IgG immunoreactivity of the sera of 20 control and 26 PXFG patients to purified C6orf129 was analysed using a reverse enzyme-linked immunoassay. The gene expression profile of C6orf129 in 40 different tissues was analysed using tissue qPCR array plates.
We detected an increased prevalence among the PXFG patients of serum antibodies to seven proteins - C6orf129; stathmin-like 4; transmembrane protein 9 domain family, member B; fibroblast growth factor receptor 3; cleft lip and palate transmembrane protein 1; EH- domain containing protein 1 and eukaryotic translation elongation factor 2. All antigens are expressed in RGC-5 cells and cDNA from human brain and optic nerve with the exception of stathmin-like 4 in the RGC-5 cells. Patients with PXFG have increased anti-C6orf129 IgG immunoreactivity compared to controls (P<0.05). C6orf129 is ubiquitously expressed in all tissue particularly in those with a secretory role.
Screening high-density protein arrays identifies unique antibody profiles that may potentially act as biomarkers for patients with PXFG. Further characterisation of this autoantibody repertoire may also provide new insights into the pathophysiology of PXFG.
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