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Basic studies for development of disease-specific therapeutic strategies against autoimmune diseases

开发针对自身免疫性疾病的疾病特异性治疗策略的基础研究

基本信息

批准号：
12470181
负责人：
NISHIKAWA Takeji
金额：
$ 10.37万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470181/
关键词：
Autoimmune Autoantibody Model mouse epitope bullous disease knockout mouse pemphigus desmoglein

项目摘要

The purpose of this study is to obtain basic knowledge for the development of disease-specific therapeutic strategies against autoimmune diseases. We used pemphigus, an autoimmune blistering diseases of the skin and mucous membranes. Patients with pemphigus vulgaris (PV) and foliaceus (PF) have circulating pathogenic IgG autoantibody against desmoglein. 3 (Dsg3) and desmoglein 1 (Dsg 1), respectively. We took two independent approaches, one was using patients specimen and the other was using a mouse model of pemphigus vulgaris. In the approach using patients specimen, we generated Dsg1- and Dsg3-domain-swapped molecules and point-mutated Dsg3 molecules with Dsg 1-specific residues by baculovirus expression to map conformational epitopes of Dsg 1 and Dsg3 in PF and PV. The binding of autoantibodies to the mutant molecules was assessed by competition ELISA. Domain-swapped molecules containing the N-terminal 161 residues of Dsg1 and Dsg3 yielded greater than 50% competition in 30/43 (69.8%) PF sera and 31/40 (77.5%) PV sera, respectively. Within these N-terminal regions, most of the epitopes were mapped to residues 26-87 of Dsg1 and 25-88 of Dsg3. These findings suggest that the dominant autoimmune epitopes in both PF and PV are found in the N-terminal adhesive surfaces of Dsgs. In the approach using the PV mouse model, we succeeded in obtaining several anti-Dsg3 mouse monoclonal IgG antibodies from the PV model mice. Among them AK19 and AK23 showed the pathogenic activity in inducing blister formation. At this point we could not obtain any specific peptides to bind these pathogenic antibodies. Through these studies we could obtain important tools to develop the disease-specific immune suppressive therapy.

本研究的目的是获得基础知识的疾病特异性治疗策略的发展对自身免疫性疾病。我们使用天疱疮，一种皮肤和粘膜的自身免疫性起泡疾病。寻常型天疱疮（PV）和落叶型天疱疮（PF）患者循环中存在抗桥粒芯糖蛋白的致病性IgG自身抗体。3（Dsg 3）和桥粒芯糖蛋白1（Dsg 1）。我们采取了两种独立的方法，一种是使用患者标本，另一种是使用寻常型天疱疮小鼠模型。在使用患者标本的方法中，我们通过杆状病毒表达产生了Dsg 1和Dsg 3结构域交换的分子和具有Dsg 1特异性残基的点突变的Dsg 3分子，以定位PF和PV中Dsg 1和Dsg 3的构象表位。通过竞争ELISA评估自身抗体与突变体分子的结合。含有Dsg 1和Dsg 3的N-末端161个残基的结构域交换分子分别在30/43（69.8%）的PF血清和31/40（77.5%）的PV血清中产生大于50%的竞争。在这些N-末端区域内，大多数表位定位于Dsg 1的残基26-87和Dsg 3的残基25-88。这些发现表明，在PF和PV中的优势自身免疫表位被发现在Dsgs的N-末端粘附表面。在使用PV小鼠模型的方法中，我们成功地从PV模型小鼠获得了几种抗Dsg 3小鼠单克隆IgG抗体。其中AK 19和AK 23表现出诱导水疱形成的致病活性。在这一点上，我们不能获得任何特定的肽结合这些致病性抗体。通过这些研究，我们可以获得重要的工具，发展疾病特异性免疫抑制治疗。