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Research for Anti-Retinal Antibody in Retinal Disorders

抗视网膜抗体在视网膜疾病中的研究

基本信息

批准号：
63480389
负责人：
NAKAZAWA Mitsuru
金额：
$ 3.9万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480389/
关键词：
Retina Retinal Disoeders Retina specific antigens Molecular Biology Retina specific cDNA clones Retinitis pigmentosa ロドプシン S抗原カテプシンD 神経網膜網膜色素上皮細胞網膜抗原抗網膜抗体

项目摘要

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.

已经表明，抗视网膜抗体的产生和影响某些种类的视网膜疾病的发病机制。在该项目中，作为第一步，定性和定量地检查是否在视网膜疾病患者的血清中检测到抗视网膜抗体。视紫红质、视网膜S-抗原、组织蛋白酶D和视网膜粗提物用作视网膜抗原。长期和广泛的孔源性视网膜脱离以及急性视网膜坏死或炎性色素性静脉旁视网膜脉络膜萎缩的患者显示高滴度的抗视网膜粗提物抗体。克隆了视紫红质、视网膜S抗原、转导素（α亚基）、IRBP、CRALBP、MEKA、蛋白和33 kDa蛋白的cDNA，并转染E.大肠杆菌JM 109与合适的质粒载体连接，从而使患者基因组DNA的分子生物学分析能够容易地作为临床部门的实验室检查进行。此外，为了准备分离未知的视网膜抗原的实验，从人脑中分离的mRNA合成cDNA，并制备人视网膜cDNA文库。作为本项目的第三步，使用聚合酶链反应和限制性片段长度多态性对视网膜色素变性患者的基因组DNA进行了检测，特别是对视紫红质基因进行了检测。结果发现，一个常染色体显性遗传的青少年视网膜炎家系中有2例患者存在视紫红质基因点突变，提示视紫红质基因可能与一种青少年视网膜炎的发病有关。