Research of disease selective immunotherapy in autoimmune diseases

自身免疫性疾病的疾病选择性免疫治疗研究

• 批准号: 09556068
• 负责人: IWASAKI Toshiroh
• 金额: $ 8.06万
• 依托单位: Tokyo University of Agriculture & Technology (1999)Gifu University (1997-1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09556068/
• 关键词: dog; pemphigus; ELISPOT; desmoglein; autoimmune disease; animal model; desmonsome; 自己免疫疾患; 天疱痘; デスモソーム

Pemphigus is a life-threatening autoimmune blistering disease of the skin and mucous membranes. In patients with pemphigus, circulating autoantibodies against cell surface of keratinocytes in stratified squamous epithelia play a pathogenic part, and cause loss of cell-cell adhesion with resultant blister formation. Since the detection of autoantibodies by indirect immunofluorescence in canine pemphigus is usually difficult probably due to the low sensitivity of the substrate, several substrates for immunofluorescence were studied. As a result, bovine esophagus showed highest sensitivity and specifity. For investigating the autoantigens directed by the serum of pemphigus foliaceus, cloning and sequencing of canine desmoglein 1 (Dsg1) were performed using cDNA library from canine skin. Fusion proteins of Dsg1 expressed in baculovirus were made. These proteins reacted with sera from human patients with pemphigus foliaceus, suggesting that canine Dsg1 crossreacted with human autoantibodies from pemphigus foliaceus. ELISPOT (enzyme-linked immunospot assay) was developed for the detection of antigen-specific B cells in patients with pemphigus. This method will provide us to analyze frequencies of Dsg-specific IgG producing and memory B cells.

天疱疮是一种危及生命的皮肤和粘膜自身免疫性起泡疾病。在天疱疮患者中，针对复层鳞状上皮细胞中角质形成细胞表面的循环自身抗体起致病作用，并导致细胞-细胞粘附丧失，从而形成水疱。由于通过间接免疫荧光法检测犬天疱疮中的自身抗体通常很困难，可能是由于底物的灵敏度低，因此研究了几种用于免疫荧光的底物。结果表明，牛食管的敏感性和特异性最高。为研究落叶型天疱疮血清中的自身抗原，利用犬皮肤cDNA文库对犬桥粒芯糖蛋白1（Dsg 1）进行了克隆和序列分析。制备了在杆状病毒中表达的Dsg 1融合蛋白。这些蛋白质与人类落叶型天疱疮患者的血清反应，表明犬Dsg 1与人类落叶型天疱疮自身抗体发生交叉反应。ELISPOT（酶联免疫斑点试验）被开发用于检测天疱疮患者的抗原特异性B细胞。该方法将为我们分析Dsg特异性IgG产生和记忆B细胞的频率提供帮助。

项目成果

Ishii K et al.: "Desmoglein 1 and desmoglein 3 are the target autoantibodies in herpetiformis pemphifgus"Arch dermatol.

Ishii K 等人：“桥粒芯糖蛋白 1 和桥粒芯糖蛋白 3 是疱疹样天疱疮中的目标自身抗体”Arch dermatol。

Amagai M: "The clinical phenotype of pemphigus is defined by the anti-desmoglein autoantibody profile"J Am Acad Dermatol. 40・2. 167-169 (1999)

Amagai M：“天疱疮的临床表型由抗桥粒芯糖蛋白自身抗体谱定义”J Am Acad Dermatol 40・2 (1999)。

Hernandez C et al.: "Pemphigus foliaceus preferential binding of IgG1 and C3 at the upper epidernis" Br J Dermatol 136. 136. 249-252 (1997)

Hernandez C 等人：“落叶型天疱疮在上表皮优先结合 IgG1 和 C3”Br J Dermatol 136. 136. 249-252 (1997)

Kubo A.et al.: "Herpetiform penphigug showing reactivity with pemphigus rulgaris antigen (desmolglein 3)" Br J Dermatol. 137. 169-113 (1997)

Kubo A.et al.：“Herpetiform penphigug 显示与寻常性天疱疮抗原（desmolglein 3）的反应性”Br J Dermatol。

Amagai M et al.: "Usefulness of enzyme-linked immunosorbent assay using recombinant desmoglein 1 and 3 for serodiagnosis of pemphigus"Br J. Dermatol.

Amagai M 等人：“使用重组桥粒芯糖蛋白 1 和 3 进行酶联免疫吸附测定对天疱疮血清诊断的有用性”Br J. Dermatol。