title: Cross-reactivity of antibodies against leptospiral recurrent uveitis-associated proteins A and B (LruA and LruB) with eye proteins.
abstract: Infection by Leptospira interrogans has been causally associated with human and equine uveitis. Studies in our laboratories have demonstrated that leptospiral lipoprotein LruA and LruB are expressed in the eyes of uveitic horses, and that antibodies directed against LruA and LruB react with equine lenticular and retinal extracts, respectively. These reactivities were investigated further by performing immunofluorescent assays on lenticular and retinal tissue sections. Incubation of lens tissue sections with LruA-antiserum and retinal sections with LruB-antiserum resulted in positive fluorescence. By employing two-dimensional gel analyses followed by immunoblotting and mass spectrometry, lens proteins cross-reacting with LruA antiserum were identified to be alpha-crystallin B and vimentin. Similarly, mass spectrometric analyses identified beta-crystallin B2 as the retinal protein cross-reacting with LruB-antiserum. Purified recombinant human alpha-crystallin B and vimentin were recognized by LruA-directed antiserum, but not by control pre-immune serum. Recombinant beta-crystallin B2 was likewise recognized by LruB-directed antiserum, but not by pre-immune serum. Moreover, uveitic eye fluids contained significantly higher levels of antiibodies that recognized alpha-crystallin B, beta-crystallin B2 and vimentin than did normal eye fluids. Our results indicate that LruA and LruB share immuno-relevant epitopes with eye proteins, suggesting that cross-reactive antibody interactions with eye antigens may contribute to immunopathogenesis of Leptospira-associated recurrent uveitis.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, USA. ashutosh.verma@uky.edu
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Verma
authors.forename: Pawan
authors.initials: P
authors.lastname: Kumar
authors.forename: Kelly
authors.initials: K
authors.lastname: Babb
authors.forename: John F
authors.initials: JF
authors.lastname: Timoney
authors.forename: Brian
authors.initials: B
authors.lastname: Stevenson
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000276:immunology; D007921:Leptospira interrogans / Q000276:immunology*; D013058:Mass Spectrometry; D012160:Retina / Q000276:immunology; D014605:Uveitis / Q000276:immunology* / Q000382:microbiology*; D014746:Vimentin / Q000276:immunology; D038203:alpha-Crystallin B Chain / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D014746:Vimentin; D038203:alpha-Crystallin B Chain