原田病の分子免疫学的研究(抗原の決定とクロ-ニングを中心として)

原田病的分子免疫学研究（重点是抗原测定和克隆）

• 批准号: 03670825
• 负责人: 山木 邦比古
• 金额: $ 1.22万
• 依托单位: Akita University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670825/
• 关键词: 原田病; 自己免疫; Molecular cloning

原因病はメラノサイドに対する自己免疫疾患であり、細胞性免疫がその主体をなすと考えられている。本研究では最初に各種melanocyte,牛ぶどう膜粗抽出液をmitogenとして、原田病患者から採取した抹消血リンパ球、及び脳脊髄液中のリンパ球を用い、 ^3Hthymidineの取り込みによるLymphocyte prolife ration assayを行った。melanocyteはGー361、WN115、Bowes、WN9、HMV1の粗抽出液可溶性成分を用いた。いづれの抗原に対しても、抹消血リンパ球、脳脊髄液ともに有意の反応を示すものはなかった。次に、これら抗原を分子量別に分画し、それぞれの分画を濃縮後、同様の方法にてassayした。この方法では分子量約7万の分画にproliferationを示す蛋白質の存在が示された。更にこの蛋白を単一の蛋白として精製分離するため二次元電気泳動を行った。等電点電気泳動ではPH約6.8の部分に増殖を示す部分があり、この部分を分子量別に二次元に展開した。展開したそれぞれのペプタイドは分子量約7万を示すものを採取し、lymphocyte proliferation assayを行った。これにより患者リンパ球が増殖を示すペプタイドが同定された。なお、コントロ-ル(negative)としては正常ヒト抹消血リンパ球を用いた。このペプタイドは現在アミノ酸分析中である。アミノ酸分析の結果に従い、これに対応するoli gonucleotideを合成し、すでに作製済みのcDNA libraryからこのペプタイドのcDNAをクロ-ニングする予定である。

The cause of the disease is known as autoimmune disease, cellular immunity, host disease, and so on. In this study, we initially tested all kinds of melanocyte, the crude extract of bovine oyster membrane was used to extract mitogen, the patients with Harada disease were used to wipe blood and eliminate blood, and in chiropractic fluid, the samples were collected and used. ^ 3Hthymidine was used to extract Lymphocyte prolife ration assay samples. The soluble components of the crude extract of melanocytelic Gram 361, WN115, Bowes, WN9 and HMV1 were used. The antigens are not affected, the blood is wiped, the balls are wiped, and the chiropractic fluid is intentionally negative. After the molecular weight of the antigen was separated, the molecular weight of the antigen was separated, and after the molecular weight of the antigen was separated, the molecular weight of the antigen was separated, and the molecular weight of the antigen was separated. After the molecular weight of the antigen was separated, the molecular weight of the antigen was separated, and the molecular weight of the antigen was analyzed by the same method. The molecular weight of the protein is about 70, 000. Proliferation shows the presence of the protein. The primary protein concentrate is separated from the second dimensional electrophoretic electrophoresis. Isoelectric electrophoretic mobility shift assay (PH) showed that the molecular weights of partial and partial molecular weights were determined by double molecular weight analysis. The molecular weight is about 70,000. The molecular weight is about 70,000. The molecular weight of lymphocyte proliferation assay is about 70000. The patient said that the colonization of the ball showed that the patient had the same diagnosis. To remove blood from the body and use the ball to remove blood. (negative). There are some problems in the acid analysis. The results of acid analysis showed that the oli gonucleotide was synthesized, and the cDNA was predetermined. The results of acid analysis showed that the results of acid analysis were satisfactory, and the results of acid analysis and acid analysis showed that the results of acid analysis, acid analysis and acid analysis.