Investigative ophthalmology & visual science · 1995
Autoimmunity to a cornea-associated stromal antigen in patients with Mooren's ulcer.
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Gottsch J D, Liu S H, Minkovitz J B, Goodman D F, Srinivasan M, Stark W J
Аннотация
PURPOSE: To purify and characterize a cornea-associated antigen (CO-Ag) and to determine antibody levels to CO-Ag in patients with Mooren's ulcer.
METHOD: Standard ion exchange and gel filtration chromatographies were used to isolate and purify CO-Ag from crude bovine stromal extracts. The serum of a patient with Mooren's ulcer, containing a high level of antibodies directed against CO-Ag, was used to monitor isolation procedures. Using this newly purified CO-Ag, an enzyme-linked immunoabsorbent assay was used to detect the presence of antibodies to CO-Ag in the sera of other patients with Mooren's ulcer.
RESULTS: CO-Ag was purified to apparent homogeneity from bovine corneal stromal extracts by a series of ion exchange chromatographies and gel filtration. Polyacrylamide gel electrophoresis showed that CO-Ag was a tetramer with a molecular weight of 30,000 d that may dissociate under denaturing conditions into a monomer of 7000 d. Strong indirect immunofluorescent staining was demonstrated of the stroma by guinea pig anti-CO-Ag antibody. A statistically significant difference in the level of specific antibodies to CO-Ag between patients with Mooren's ulcer and controls was found (P < 0.001). The antibody level was elevated in patients with Mooren's ulcer (mean antibody level, 0.58 +/- 0.13) compared with the controls (mean antibody level, 0.22 +/- 0.04).
CONCLUSION: These results suggest that an autoantigen exists in the corneal stroma that reacts with serum antibodies from patients with Mooren's ulcer. The availability of a purified corneal antigen could facilitate the diagnosis and define the pathogenetic mechanisms in Mooren's ulcer.
METHOD: Standard ion exchange and gel filtration chromatographies were used to isolate and purify CO-Ag from crude bovine stromal extracts. The serum of a patient with Mooren's ulcer, containing a high level of antibodies directed against CO-Ag, was used to monitor isolation procedures. Using this newly purified CO-Ag, an enzyme-linked immunoabsorbent assay was used to detect the presence of antibodies to CO-Ag in the sera of other patients with Mooren's ulcer.
RESULTS: CO-Ag was purified to apparent homogeneity from bovine corneal stromal extracts by a series of ion exchange chromatographies and gel filtration. Polyacrylamide gel electrophoresis showed that CO-Ag was a tetramer with a molecular weight of 30,000 d that may dissociate under denaturing conditions into a monomer of 7000 d. Strong indirect immunofluorescent staining was demonstrated of the stroma by guinea pig anti-CO-Ag antibody. A statistically significant difference in the level of specific antibodies to CO-Ag between patients with Mooren's ulcer and controls was found (P < 0.001). The antibody level was elevated in patients with Mooren's ulcer (mean antibody level, 0.58 +/- 0.13) compared with the controls (mean antibody level, 0.22 +/- 0.04).
CONCLUSION: These results suggest that an autoantigen exists in the corneal stroma that reacts with serum antibodies from patients with Mooren's ulcer. The availability of a purified corneal antigen could facilitate the diagnosis and define the pathogenetic mechanisms in Mooren's ulcer.