Epidermolysis bullosa acquisita sera react with distinct epitopes on the NC1 and NC2 domains of type VII collagen: study using immunoblotting of domain-specific recombinant proteins and postembedding immunoelectron microscopy
Epidermolysis bullosa acquisita sera react with distinct epitopes on the NC1 and NC2 domains of type VII collagen: study using immunoblotting of domain-specific recombinant proteins and postembedding immunoelectron microscopy
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DOI：
10.1111/j.1365-2133.2004.05933.x
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发表时间：
2004-05-01
影响因子：
10.3
通讯作者：
Hashimoto, T
中科院分区：
文献类型：
--
作者：
Ishii, N;Yoshida, M;Hashimoto, T
背景 大疱性表皮松解症 (EBA) 血清与 VII 型胶原发生反应，VII 型胶原是锚定原纤维的主要成分，其中主要表位被认为存在于 N 端非胶原 (NC) 1 结构域中。目的 确定 C 端 NC2 结构域中是否也存在表位，并确定其超微结构定位。 使用 VII 型胶原蛋白 NC1 和 NC2 结构域的重组蛋白进行免疫印迹，并进行包埋后免疫电子显微镜检查。 结果 测试的 28 份 EBA 血清中有 20 份与 NC1 结构域发生反应，八份血清与 NC2 结构域发生反应。与 NC1 结构域反应的血清在致密层内显示出免疫反应性，与 NC2 结构域反应的血清在致密层下方 300-360 nm 的真皮中显示出免疫反应性。 结论 这项研究清楚地鉴定了 NC2 结构域中表位的存在，并表明 NC1 结构域中的表位存在于致密层中 densa 中的表位位于致密层下方的真皮中。
Background The sera of epidermolysis bullosa acquisita (EBA) react with type VII collagen, a major component of anchoring fibrils, in which the major epitopes have been considered to be present in the N-terminal noncollagenous (NC) 1 domain.Objectives To determine whether there are also epitopes in the C-terminal NC2 domain, and to determine their ultrastructural localization.Methods Immunoblotting using recombinant proteins of the NC1 and NC2 domains of type VII collagen, and postembedding immunoelectron microscopy.Results Twenty of 28 EBA sera tested reacted with the NC1 domain and eight sera reacted with the NC2 domain. The sera that reacted with the NC1 domain showed immunoreactivity within the lamina densa and the sera that reacted with the NC2 domain showed immunoreactivity in the dermis 300-360 nm below the lamina densa.Conclusions This study clearly identified the presence of epitopes in the NC2 domain, and showed that the epitope in the NC1 domain is present in the lamina densa and that the epitope in the NC2 domain is in the dermis below the lamina densa.