Comparative study of autoantigens for various bullous skin diseases by immunoblotting using different dermo-epidermal separation techniques
Comparative study of autoantigens for various bullous skin diseases by immunoblotting using different dermo-epidermal separation techniques
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DOI：
10.1111/j.1365-2230.1995.tb01376.x
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发表时间：
1995-11-01
影响因子：
4.1
通讯作者：
Nishikawa, T
中科院分区：
文献类型：
--
作者：
Ohata, Y;Hashimoto, T;Nishikawa, T
我们研究了寻常型天疱疮（PV），增殖性天疱疮，落叶型天疱疮（Pf），巴西Pf，大疱性类天疱疮（BP），和获得性大疱性表皮细胞（EBA）血清与免疫印迹分析，使用人表皮和真皮提取物作为抗原来源的反应性。为了获得表皮和真皮提取物，使用三种不同的真皮-表皮分离方法：即，乙二胺四乙酸（EDTA）分离、热分离和分散酶分离。所有的15 PV和七个增殖性天疱疮血清表现出130 kDa的PV抗原在表皮提取物中获得的所有三种方法。此外，三个PV血清也显示了160-kDa的Pf抗原，桥粒芯糖蛋白。10个13 Pf血清和SIS的15巴西Pf血清与桥粒芯糖蛋白在相同的模式在所有三个表皮提取物。15的22个BP血清表现出反应性与230 kDa的BP抗原在所有三个表皮提取物中的相同模式，而14个BP血清检测到180 kDa的BP抗原的EDTA和热分离的表皮提取物，但不是在分散酶分离的表皮提取物。通过EDTA和热分离的真皮获得皮肤提取物，并且在两种样品中所有六种EBA血清标记290-kDa EBA抗原。这些结果表明，热分离的皮肤与EDTA分离的皮肤一样可用于检测各种自身抗原，但热分离是优选的，因为制备时间较短。
We investigated the reactivity of pemphigus vulgaris (PV), pemphigus vegetans, pemphigus foliaceus (Pf), Brazilian Pf, bullous pemphigoid (BP), and epidermolysis bullosa acquisita (EBA) sera with an immunoblot analysis using human epidermal and dermal extracts as a source of antigen. To obtain epidermal and dermal extracts three different dermo-epidermal separation methods were used: namely, ethylenediaminetetraacetic acid (EDTA) separation, heat separation, and dispase separation. All the 15 PV and the seven pemphigus vegetans sera demonstrated a 130-kDa PV antigen in epidermal extracts obtained by all the three methods. Furthermore, three PV sera also showed a 160-kDa Pf antigen, desmoglein. Ten of 13 Pf sera and sis of 15 Brazilian Pf sera reacted with desmoglein in the same pattern in all the three epidermal extracts. Fifteen of the 22 BP sera showed reactivity with 230-kDa BP antigen in the same pattern in all the three epidermal extracts, whereas 14 BP sera detected the 180-kDa BP antigen in extracts of EDTA- and heat-separated epidermis but not in dispase-separated epidermal extract. Dermal extracts were obtained by EDTA- and heat-separated dermis, and all six EBA sera labelled a 290-kDa EBA antigen in both samples. These results suggest that heat-separated skin is as useful as EDTA-separated skin for detecting various autoantigens, but heat separation is preferable because the preparation time is shorter.