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(Investigative Ophthalmology and Visual Science. 1999;40:2147-2151.)
© 1999 by The Association for Research in Vision and Ophthalmology, Inc.

Corneal Epithelial–Specific Cytokeratin 3 is an Autoantigen in Wegener’s Granulomatosis–Associated Peripheral Ulcerative Keratitis

Irena Reynolds1, Andrew B. Tullo2, Sally L. John3, P. J. Lennox Holt1 and M. Chantal Hillarby1

1 From the Musculoskeletal Research Group, 2 University Department of Ophthalmology, and 3 ARC ERU, University of Manchester, and Central Manchester Health Care NHS Trust, Manchester, United Kingdom.

Abstract

PURPOSE. In a previous investigation it was demonstrated that circulating antibodies to a 66-kDa corneal epithelial antigen (BCEA-A) are associated with peripheral ulcerative keratitis (PUK) in patients with Wegener’s granulomatosis (WG). The aim of this study was to identify BCEA-A.

METHODS. The 66-kDa antigen was purified from a bovine corneal epithelial protein extract, using DE52 ion exchange chromatography. Purified protein was used to raise rabbit polyclonal antibodies. These antibodies were used to screen a bovine corneal epithelial cDNA expression library. Positive clones were purified and sequenced. Clones were identified by DNA sequence homology searches of the GenBank DNA database.

RESULTS. A cDNA clone that demonstrated strong binding to both the rabbit polyclonal antibody and patient sera, showed 85% homology to rabbit cytokeratin 3 (K3). K3 is a basic cytokeratin specific to corneal epithelium. No bovine DNA sequence for K3 is available. However, bovine K3 is larger than rabbit K3, with a molecular weight of 66 kDa. Immunofluorescence using both patient sera and the rabbit antibody demonstrated a cytoplasmic binding pattern on human corneal epithelium.

CONCLUSIONS. This evidence suggests that the 66-kDa autoantigen (BCEA-A) associated with PUK in WG is cytokeratin 3, and this may form the basis of a diagnostic/prognostic test.




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Copyright © 1999 by the Association for Research in Vision and Ophthalmology