Development of novel therapy against bullous diseases

这一研究在团体之间开展了一项增强的国际合作，该研究将导致研究皮肤学领域的自动免疫和继承的大疾病。在这项研究中，一种新型的活性疾病模型被开发出来。敲老鼠不能获得不合格基因产品的耐受性。使用knockout macking desmoglein 3 （Dsg 3），以pemphigus vulgaris抗原（PV）为目标，我们已经建立了一种方法来生成这种自动抗体介导的疾病的活性疾病模型。Dsg 3-/-D1 mice，但不是Dsg 3-D1+/-D1 littermates，生产抗Dsg 3 IgG，如果可以与重新组合鼠标Dsg 3绑定本机Dsg 3。Splenocytes from the immunized Dsg3--D1 mice were then adoptively transferred into Rag-2-D1--D1 immunodeficient mice expressing Dsg3。反Dsg 3 IgG抗体在6个月内稳定地生产在重复老鼠而没有进一步的助推。此IgG绑定到Dsg 3在体内，并破坏了细胞-细胞的粘附。顺便说一下，在PV的典型历史学发现中，使用其口腔模糊的膜进行了繁殖。作为一种创新的治疗方法，我们已经开发了化学分子，以确定PV中的抗原特异性B细胞。用Dsg 3可以消除Dsg 3特异性杂交瘤细胞或用细胞数目减少60%的免疫小鼠产生抗Dsg 3 IgG B细胞的重组毒素。由于遗传的皮肤疾病，我们研究了突变类型和表型在显性和继发性代谢性表皮聚胞菌中的相关性。我们还制定了一个基本的协议，用于临床应用的表皮表格的生产。Several recombinant adenoviruses were developed to introduce exogenous genes to epidermal cells。

This study enhanced international collaboration between groups which lead the fields of investigative dermatology on autoimmune and inherited bullous diseases. In this study, a novel active disease model for pemphigus has been developed. Knockout mice do not acquire tolerance of the defective gene product. Using knockout mice lacking desmoglein 3 (Dsg3), the target antigen of pemphigus vulgaris (PV), we established a method to generate an active disease model for this autoantibody-mediated disease. Dsg3ィイD1-/-ィエD1 mice, but not Dsg3ィイD1+/-ィエD1 littermates, produced anti-Dsg3 IgG that was able to bind the native Dsg3 when immunized with recombinant mouse Dsg3. Splenocytes from the immunized Dsg3ィイD1-/-ィエD1 mice were then adoptively transferred into Rag-2ィイD1-/-ィエD1 immunodeficient mice expressing Dsg3. Anti-Dsg3 IgG was stably produced in the recipient mice for over 6 months without further boosting. This IgG bound to Dsg3 in vivo and disrupted the cell-cell adhesion of keratinocytes. Consequently, the recipient mice developed erosions in their oral mucous membranes with typical histologic findings of PV. As an innovative therapeutic approach, we have developed chimeric molecules for targeting of antigen-specific B cells in PV. The recombinant toxins fused with Dsg3 could eliminate Dsg3-specific hybridoma cells or anti-Dsg3 IgG producing B cells from immunized mice with a 60% reduction in cell number. For inherited skin diseases, we investigated the correlation between the type of mutations and the phenotype in dominant and recessive dystrophic epidermolysis bullosa. We also set up basic protocol for production of epidermal sheets for clinical application. Several recombinant adenoviruses were developed to introduce exogenous genes to epidermal cells.