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Development of novel therapy against bullous diseases

针对大疱性疾病的新疗法的开发

基本信息

批准号：
10044318
负责人：
NISHIKAWA Takeji
金额：
$ 5.44万
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：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10044318/
关键词：
Autoimmune Inherited skin disease Desmoglein Laminin 5 Type VII collagen Animal model Epidermal sheets Gene therapy ノックアウトマウス遺伝子解析自己免疫皮膚疾患

项目摘要

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.

这项研究加强了领导自身免疫性和遗传性大疱性疾病皮肤病学研究领域的团体之间的国际合作。在这项研究中，一个新的活动性疾病模型天疱疮已经开发。敲除小鼠不能获得对缺陷基因产物的耐受性。使用缺乏桥粒芯糖蛋白3（Dsg 3），寻常型天疱疮（PV）的靶抗原的基因敲除小鼠，我们建立了一种方法来产生这种自身抗体介导的疾病的活动性疾病模型。当用重组小鼠Dsg 3免疫时，Dsg 3单克隆D1-/-单克隆D1小鼠而不是Dsg 3单克隆D1+/-单克隆D1同窝出生的小鼠产生能够结合天然Dsg 3的抗Dsg 3 IgG。然后将来自免疫的Dsg 3 β D1-/-β D1小鼠的脾细胞过继转移到表达Dsg 3的Rag-2 β D1-/-β D1免疫缺陷小鼠中。抗Dsg 3 IgG在受体小鼠中稳定产生超过6个月，无需进一步加强。该IgG在体内与Dsg 3结合并破坏角质形成细胞的细胞-细胞粘附。因此，受体小鼠的口腔粘膜出现糜烂，具有典型的PV组织学发现。作为一种创新的治疗方法，我们已经开发了嵌合分子，用于靶向PV中的抗原特异性B细胞。与Dsg 3融合的重组毒素可清除免疫小鼠的Dsg 3特异性杂交瘤细胞或产生抗Dsg 3 IgG的B细胞，细胞数量减少60%。对于遗传性皮肤病，我们研究了显性和隐性营养不良性大疱性表皮病的突变类型和表型之间的相关性。我们还建立了用于临床应用的表皮片生产的基本方案。几种重组腺病毒被开发用于将外源基因导入表皮细胞。