Pathogenic analysis for autoimmune and hereditary skin diseases due to abnormality of desmosomes

通过使用转染了人桥粒柯林斯1-3的cDNA的COS-7细胞的免疫荧光，我们已经表明，一些天疱疮血清含有与桥粒柯林斯反应的伊加抗体。伊加天疱疮有两种亚型：IEN型和SPD型。通过免疫电镜研究，我们发现SPD型伊加天疱疮的伊加抗体与桥粒区域反应，而IEN型伊加天疱疮的伊加抗体与非桥粒区域反应。我们已经表明，大多数IgG/伊加天疱疮血清中有IgG和伊加抗体与Dsg 1/Dsg 3和Dsc 1反应。此外，免疫电镜研究表明，这些血清与桥粒区反应。我们检查了一个大家族，患者在基底层以上显示棘层松解性水疱。首先，我们检测了桥粒芯糖蛋白3基因，但我们没有发现突变。然而，我们发现了一个突变的ATP 2A 2基因，编码SERCA 2，Darier病的致病基因。在一个病人显示浅表水疱形成的表皮，我们还没有检测到任何突变的基因桥粒蛋白。相反，在这个病人中，我们发现角蛋白1基因的C-末端区域发生了突变。我们已经生产了各种重组蛋白的envoplakin和periplakin。通过免疫印迹使用这些重组蛋白，我们已经表明，大多数副肿瘤性天疱疮患者反应的不同领域的envoplakin和periplakin。我们已经成功地产生了桥粒蛋白基因敲除小鼠。桥粒未见形态学异常。此外，。来自敲除小鼠的细胞培养物没有显示出异常的细胞粘附。

By immunofluorescence using COS-7 cells transfected with cDNAs of human desmocollins 1-3, we have shown that some pemphigus sera contained IgA antibodies reactive with desmocollins. There are two subtypes of IgA pemphigus, IEN type and SPD type. By immuno-electron microscopic study, we have shown that IgA antibodies of SPD type IgA pemphigus reacted with desmosomal areas, whereas IgA antibodies of IEN type IgA pemphigus reacted with non-desmosomal areas. We have shown that most of the sera of IgG/IgA pemphigus had IgG and IgA antibodies reactive with Dsg1/Dsg3 and Dsc1. In addition, by immuno-electron microscopic study, it is shown that these sera reacted with desmosomal areas. We have examined a large family with patients showing acantholytic blister above the basal layer. First, we examined desmoglein 3 gene, but we could not find a mutation. However, we have found a mutation in the ATP2A2 gene, encoding SERCA2, the causative gene of Darier disease. In a patient showing superficial blister formation in the epidermis, we have not detected any mutations in the genes of desmosomal proteins. In stead, in this patient, we have found a mutation in the C-terminal area of keratin 1 gene. We have produced various recombinant proteins of envoplakin and periplakin. By immunoblotting using theses recombinant proteins, we have shown that most paraneoplastic pemphigus patients reacted with various domains of envoplakin and periplakin. We have succeeded to produce knockout mice of desmoyokin. We could not find any morphological abnormality in the desmosomes. In addition,. the cell culture from the knockout mice did not show an abnormal cell adhesion.