title: Characterization of two corneal epithelium-derived antigens associated with vasculitis.
abstract: OBJECTIVE
  In a previous investigation into corneal autoimmunity, it was demonstrated that a putative autoantigen, a protein of 66 kDa, present in bovine corneal epithelium, binds circulating autoantibodies in approximately 60% of patients with Wegener's granulomatosis (WG). The aim of the present study was to characterize and identify the 66-kDa protein.
  
  
  METHODS
  A purification protocol was established for the 66-kDa protein using standard chromatography techniques. During the purification procedure it became clear that the 66-kDa protein detected in patients' sera was in fact two proteins, both running at 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, that eluted in different fractions on DE-52 chromatography columns. These two proteins have been labeled bovine corneal epithelial antigen-A and -B (BCEA-A and BCEA-B). Further investigations of antibody binding have demonstrated that patients' sera bind to either one or the other of these proteins with no cross-reactivity between them. Separated BCEA-A and BCEA-B protein extracts were immunoblotted with 27 WG patients' sera, 10 Churg-Strauss syndrome (CSS) patients' sera, 31 rheumatoid arthritis (RA) patients' sera, and 40 healthy control subjects' sera from the blood bank.
  
  
  RESULTS
  Forty-six percent of WG patients' sera had antibodies to one of the 66-kDa antigens, whereas none of the healthy control subjects' sera had 66-kDa antibodies (P < 10(-5)). In the WG group, 31% were positive to BCEA-A (versus controls, P = 0.0023), and 15% were positive to BCEA-B. WG patients with peripheral ulcerative keratitis (PUK) had a significant association with anti-BCEA-A antibodies when compared with healthy control subjects (50%, P < 10(-6)). However, in the RA group with no eye disease there was an association with BCEA-A (25%, P = 0.011) but not in the RA group with PUK. The frequency of anti-BCEA-B antibodies was significantly increased in patients with CSS (60%, P < 10(-7)).
  
  
  CONCLUSIONS
  In summary, it has been shown that vasculitis patients have antibodies to two 66-kDa corneal antigens and that autoantibodies to these antigens are mutually exclusive. It has also been shown that antibodies to BCEA-B are associated with CSS, whereas BCEA-A antibodies are associated with WG and RA.
authors.affiliation: Department of Rheumatology, University of Manchester, UK.
authors.forename: I
authors.initials: I
authors.lastname: Reynolds
authors.forename: S L
authors.initials: SL
authors.lastname: John
authors.forename: A B
authors.initials: AB
authors.lastname: Tullo
authors.forename: S
authors.initials: S
authors.lastname: Ayad
authors.forename: K
authors.initials: K
authors.lastname: Morgan
authors.forename: F W
authors.initials: FW
authors.lastname: Ballardie
authors.forename: P J
authors.initials: PJ
authors.lastname: Holt
authors.forename: M C
authors.initials: MC
authors.lastname: Hillarby
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001172:Arthritis, Rheumatoid / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D001323:Autoantibodies / Q000032:analysis*; D001324:Autoantigens / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003231:Conjunctivitis / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D003320:Corneal Ulcer / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional; D019573:Epithelium, Corneal / Q000737:chemistry / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting; D008875:Middle Aged; D008970:Molecular Weight; D011817:Rabbits; D014657:Vasculitis / Q000276:immunology / Q000378:metabolism* / Q000473:pathology
chemical_list: D001323:Autoantibodies; D001324:Autoantigens