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Irena Reynolds, Andrew B. Tullo, Sally L. John, P. J. Lennox Holt, M. Chantal Hillarby; Corneal Epithelial–Specific Cytokeratin 3 is an Autoantigen in Wegener’s Granulomatosis–Associated Peripheral Ulcerative Keratitis. Invest. Ophthalmol. Vis. Sci. 1999;40(9):2147-2151.
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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
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
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
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