权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Study on pathophysiological mechanism of autoantibody production in pemphigus

天疱疮自身抗体产生的病理生理机制研究

基本信息

批准号：
11470185
负责人：
AMAGAI Masayuki
金额：
$ 9.6万
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：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470185/
关键词：
autoimmune diseases pemphigus animal model desmoglein autoantibody tolerance B cell knockout mouse モデルハウス

项目摘要

Mechanisms of tolerance break against desmoglein 3 (Dsg3) in patients with pemphigus vulgaris (PV) producing pathogenic anti-Dsg3 IgG autoantibodies are unclear. In this study, we had dual approaches using human materials of PV patients and a mouse model for PV.To examine cellular mechanisms underlying the autoantibody production in PV patients, we have successfully developed an Enzyme-Linked Immunospot (ELISPOT) assay which was able to detect Dsg3-specific autoimmune B cells quantitatively. The in vitro anti-Dsg3 Ab production was abolished when CD4^+ cells were depleted or when anti-HLA-DR or anti-HLA-DQ monoclonal Ab was added to the cultures, suggesting the important role of HLA class II-restricted CD4^+ T cells in the autoAb production in PV.Using a novel PV mouse model involving Dsg3 knockout mice, we investigated the mechanisms of tolerance loss against Dsg3. Adoptive transfer of Dsg3^<-/-> splenocytes immunized with recombinant mouse Dsg3 to Rag2^<-/-> recipient mice expressing Dsg3 resulted in the stable production of anti-Dsg3 IgG and development of PV phenotypes including oral erosions with suprabasilar acantholysis. When purified T and B cells from Dsg3^<-/->, Dsg3^<+/-> or Dsg3^<+/+> mice were mixed with various combinations and transferred to Rag2^<-/-> mice, pathogenic anti-Dsg3 IgG production was observed only with a combination of Dsg3^<-/-> T and Dsg3^<-/-> B cells but not with the other combinations. These results suggest that loss of tolerance against Dsg3 in both B and T cells is important for the development of autoimmune state of PV.

寻常型天疱疮（PV）患者对桥粒芯糖蛋白3（Dsg 3）的耐受性破坏机制尚不清楚，该患者产生致病性抗Dsg 3 IgG自身抗体。在这项研究中，我们有双重的方法，使用人类材料的PV患者和小鼠模型PV。为了研究细胞机制的PV患者的自身抗体的生产，我们已经成功地开发了酶联免疫斑点（ELISPOT）检测，能够定量检测Dsg 3特异性自身免疫B细胞。当去除CD 4 ^+细胞或加入抗HLA-DR或抗HLA-DQ单克隆抗体时，体外抗Dsg 3抗体的产生被消除，表明HLA II类限制性CD 4 ^+ T细胞在PV自身抗体产生中的重要作用。我们使用一种新的PV小鼠模型，包括Dsg 3基因敲除小鼠，研究了对Dsg 3耐受性丧失的机制。将用重组小鼠Dsg 3免疫的Dsg 3 ^<-/->脾细胞连续转移到表达Dsg 3的Rag 2 ^<-/->受体小鼠，导致抗Dsg 3 IgG的稳定产生和PV表型的发展，包括口腔糜烂伴基底上棘层松解。当将来自Dsg 3 ^<-/->、Dsg 3 ^<+/->或Dsg 3 ^<+/+>小鼠的纯化的T和B细胞与各种组合混合并转移至Rag 2 ^<-/->小鼠时，仅在Dsg 3 ^<-/-> T和Dsg 3 ^<-/-> B细胞的组合中观察到致病性抗Dsg 3 IgG产生，而在其它组合中未观察到。这些结果表明，在B和T细胞中对Dsg 3的耐受性的丧失对于PV的自身免疫状态的发展是重要的。