In vitro and rapid in situ transglutaminase assays for congenital ichthyoses -: A comparative study
In vitro and rapid in situ transglutaminase assays for congenital ichthyoses -: A comparative study
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DOI：
10.1046/j.1523-1747.1998.00132.x
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发表时间：
1998-03-01
影响因子：
6.5
通讯作者：
Huber, M
中科院分区：
文献类型：
--
作者：
Hohl, D;Aeschlimann, D;Huber, M
常染色体隐性遗传先天性鱼鳞病是一种异质性的毁容性皮肤病。它们的一般特征是可变的结垢和红皮病，患者在出生时经常是胶体婴儿。常染色体隐性遗传的先天性鱼鳞病在我们的50个家族中有25个是由角质细胞转谷氨酰胺酶（TGK）缺陷引起的。常染色体隐性遗传先天性鱼鳞病的病因分类在临床上是困难的，而对于胶体婴儿则是不可能的。因此，我们在冷冻皮肤切片上建立了一种快速的TGK原位检测方法，结合免疫组织化学检测TGK蛋白，加入丹酰尸胺来评估转谷氨酰胺酶（TG）的活性。结果与培养的分化角质形成细胞的TG活性水平进行比较。十六26患者,包括胶宝贝,有强烈降低TG活动区分细胞外围的角化细胞和膜结合体外TG活动,从2.2到281.3 pmol / h毫克。九26患者,包括胶婴儿,表现出强烈的TG活动区分细胞外围的角化细胞原位体外和膜结合TG活动范围1519到10917 pmol / h毫克。在一个案例中,TG测定原位模棱两可;然而，体外膜TG活性非常低，为76.9 pmol/h X mg。我们的结果表明，体外和原位TG检测具有良好的相关性。此外，我们提出了一个新的测试与预后价值的胶凝婴儿表型。该检测允许常染色体隐性先天性鱼鳞病的快速病原分类，只有一个警告，即在罕见的模棱两可的情况下，可能有必要进行groper分类，以评估体外膜结合的TG活性。
Autosomal recessive congenital ichthyoses are a heterogeneous group of disfiguring skin diseases. They are generally characterized by variable scaling and erythroderma, and patients are frequently collodion babies at birth. Autosomal recessive congenital ichthyoses are represented in 25 of our 50 families by a defective keratinocyte transglutaminase (TGK). Pathogenic classification is difficult to assess on clinical grounds for autosomal recessive congenital ichthyoses and impossible for collodion babies. Thus, we have established a rapid TGK assay in situ on frozen skin sections using incorporation of dansyl-cadaverin to assess transglutaminase (TG) activity in combination with immunohistochemistry for TGK protein. Results were compared with TG activity levels measured in cultured differentiating keratinocytes. Sixteen of 26 patients, including a collodion baby, had strongly diminished TG activity in the cell periphery of differentiating keratinocytes and membrane-bound TG activities in vitro, ranging from 2.2 to 281.3 pmol per h mg. Nine of 26 patients, including a collodion baby, showed strong TG activity in the cell periphery of differentiating keratinocytes in situ and membrane-bound TG activities in vitro ranged front 1519 to 10917 pmol per h mg. In one case, TG assay in situ was ambiguous; however, membranous TG activity in vitro was very low at 76.9 pmol/h X mg. Our results demonstrate an excellent correlation of TG assays in vitro and in situ. In addition, we present a novel test with prognostic value for the collodion baby phenotype. This assay allows rapid pathogenic classification of autosomal recessive congenital ichthyoses with only one caveat that ill rare ambiguous cases it might be necessary for groper classification to assess membrane-bound TG activity in vitro.