| Research Abstract |
It has been suggested that anti-retina antibodies are produced and affect the pathogenesis of some kinds of retinal disorders. In this project, as the first step, it was examined whether or not anti-retina antibodies were detected in the serum of patients with retinal disorders qualitatively and quantitatively. Rhodopsin, retinal S-antigen, cathepsin D and crude extract retina were used as retinal antigens. Patients with prolonged and extensive rhegmatogenous retinal detachment as well as those with acute retinal necrosis or inflammatory pigmented paravenous retinochoroidal atrophy exhibited high titers of antibody to retinal crude extract.
As the second step of this project, retina specific antigens were examined. Cloned cDNA's for rhodopsin, retinal S-antigen, transducin (alpha-subunit), IRBP, CRALBP, MEKA, protein and 33kDa protein were obtained and transfected into E. coli JM109 with approproiate plasmid vectors, so that molecular biological analysis of patients' genomic DNA is able to be readily performed as a laboratory examination in a clinical department. Moreover, in order to prepare experiments for the isolation of unknown retinal antigens, cDNA was synthesized from mRNA isolated from human brains and human retinal cDNA library was prepared.
As the third step of this project, genomic DNA's from patients with retinitis pigmentosa were examined with a particular interest on the rhodopsin gene using polymerase chain reaction and restriction fragment length polymorphism. As a result, two cases of one pedigree with autosomal dominant retinitis pimentosa was found to have a point mutation in the rhodopsin gene, suggesting that rhodopsin gene could be related to the pathogenesis of a kind of retinitis pimentosa.
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