Epitope mapping of anti-rhodopsin antibodies from patients with normal pressure glaucoma.
OBJECTIVE
The presence of anti-rhodopsin antibodies in patients with normal pressure glaucoma (NPG) has been previously demonstrated with western blot analysis and enzyme-linked immunosorbent assay. To learn more about the characteristics, origin, and possible significance of these antibodies, the epitopic specificity of the anti-rhodopsin antibodies was examined in four NPG patients.


METHODS
Antibodies in patient sera were assayed by western blot analysis against purified bovine rhodopsin. Peptides derived from particular segments of the rhodopsin sequence were tested for activity in competing for rhodopsin-antibody binding.


RESULTS
Of a series of nine peptides that constitute most of the hydrophilic regions of rhodopsin, only one, consisting of the C-terminal 25 amino acids, prevented binding of the patient antibodies to rhodopsin. Higher resolution mapping using a set of dodecamers of overlapping sequences from the C-terminal region demonstrated that antibody binding is completely dependent on the last two amino acids. Removing the C-terminal alanine alone, or amidating the C terminus carboxyl group, also eliminated antibody binding.


CONCLUSIONS
Because four of four patient antibodies examined exhibited the identical epitopic specificity, it is likely that a common mechanism underlies their generation. This may indicate that molecular mimicry has occurred, because several pathogens contain similar C-terminal sequences. Although they may serve as diagnostic markers, and provide evidence that there is an autoimmune component in some patients with glaucoma, the role, if any, that these antibodies play in the pathogenesis of the disease remains unknown.
Romano
C
C
Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Li
Z
Z
Arendt
A
A
Hargrave
P A
PA
Wax
M B
MB
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D002417:Cattle; D000939:Epitopes / Q000276:immunology*; D005901:Glaucoma / Q000276:immunology* / Q000503:physiopathology*; D006801:Humans; D007429:Intraocular Pressure / Q000502:physiology*; D012016:Reference Values; D012243:Rhodopsin / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D012243:Rhodopsin