Research for Anti-Retinal Antibody in Retinal Disorders

抗视网膜抗体的产生影响了一些视网膜疾病的发病机制。本项目首先对视网膜疾病患者血清中是否检测到抗视网膜抗体进行定性和定量的检测。视紫红质、视网膜s抗原、组织蛋白酶D和视网膜粗提物作为视网膜抗原。长期和广泛的孔源性视网膜脱离患者以及急性视网膜坏死或炎症性色素静脉旁视网膜脉络膜萎缩患者对视网膜粗提物的抗体滴度高。作为该项目的第二步，检测视网膜特异性抗原。获得了克隆的视紫红质、视网膜s抗原、转导素（α亚基）、IRBP、CRALBP、MEKA、蛋白和33kDa蛋白的cDNA，用合适的质粒载体转染到大肠杆菌JM109中，使患者基因组DNA的分子生物学分析能够方便地作为临床部门的实验室检查。此外，为了准备分离未知视网膜抗原的实验，我们从人脑中分离的mRNA合成了cDNA，并建立了人视网膜cDNA文库。作为该项目的第三步，我们使用聚合酶链反应和限制性片段长度多态性技术对色素性视网膜炎患者的基因组DNA进行了特别感兴趣的视紫红质基因检测。结果发现2例常染色体显性视网膜炎一个家系的视紫红质基因存在点突变，提示视紫红质基因可能与一类视网膜炎的发病机制有关。

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.