cross-reactivity of antibodies against leptospiral recurrent uveitis-associated proteins a and b (lrua and lrub) with eye proteins大钩端螺旋体抗体复发uveitis-associated蛋白a和b(lrua和lrub)与蛋白质.pdfVIP

Cross-Reactivity of Antibodies against Leptospiral Recurrent Uveitis-Associated Proteins A and B (LruA and LruB) with Eye Proteins 1 2¤a 1¤b 2 1 Ashutosh Verma *, Pawan Kumar , Kelly Babb , John F. Timoney , Brian Stevenson 1 Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, United States of America, 2 Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, United States of America 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 a-crystallin B and vimentin. Similarly, mass spectrometric analyses identified b- crystallin B2 as the retinal protein cross-reacting with LruB-antiserum. Purified recombinant human a-crystallin B and vimentin were recognized by LruA-directed antiserum, but not by control pre-immune serum. Recombinant b-crystallin B2 was likewise recognized by LruB-directed antiserum, but not by pre-immune serum. Moreover, uveitic eye fluids contained significan