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.

已经提出，产生抗逆转录抗体并影响某些视网膜疾病的发病机理。在该项目中，作为第一步，在定性和定量性视网膜疾病的患者的血清中是否检测到抗逆转抗体。视紫红素，视网膜S抗原，组织蛋白酶D和粗提取物视网膜用作视网膜抗原。长时间且广泛的类风湿性视网膜脱离以及患有急性视网膜坏死或炎性色素色素性偏红色视网膜类染色性萎缩的患者表现出对视网膜原油提取物的抗体。克隆的cDNA用于视紫红素，视网膜S-抗原，透明蛋白（alpha-subunit），IRBP，Cralbp，Meka，蛋白质和33KDA蛋白，并转染了大肠杆菌JM109，并具有质质质粒载体，因此，该质粒质体的分子生物学分析，因此DNA很容易在临床部门作为实验室检查。此外，为了准备分离未知视网膜抗原的实验，从从人的大脑中分离的mRNA中合成cDNA并制备了人类视网膜cDNA文库。这是该项目的第三步，从具有视网膜炎的患者的基因组DNA中，研究了。使用聚合酶链反应和限制性片段长度多态性对视紫红质基因的特别兴趣。结果，发现两例具有常染色体显性视网膜炎的pimentosa的谱系在视紫红质基因中具有一个点突变，这表明若订光蛋白基因可能与Pimentosa的一种视网膜炎的发病机理有关。