Department of Dermatology, University of Lübeck, 23538 Lübeck, Germany.
J Immunol. 2013 Sep 15;191(6):2978-88. doi: 10.4049/jimmunol.1300310. Epub 2013 Aug 19.
In autoimmune bullous dermatoses (AIBD), autoantibodies induce blisters on skin or mucous membranes, or both. Mechanisms of continued autoantibody production and blistering have been well characterized using AIBD animal models. Mechanisms leading to the initial autoantibody production, however, have not been investigated in detail. Epidermolysis bullosa acquisita (EBA) is an AIBD associated with autoantibodies to type VII collagen (COL7). The majority of EBA patients' sera recognize the noncollagenous domain 1, including the von Willebrand factor A-like domain 2 (vWFA2). In experimental EBA induced by immunization with GST-COL7, disease manifestation depended on the genetic background, a Th1 polarization, and the GST-tag. In this model, nude mice neither produced autoantibodies nor blisters. It has remained uncertain which APC and T cell subsets are required for EBA induction. We established a novel EBA model by immunization with vWFA2 fused to intein (lacking the GST-tag). All tested mouse strains developed autoantibodies, but blisters were exclusively observed in mice carrying H2s. In immunized mice, CD4 T cells specific for vWFA2 were detected, and their induction required presence of B cells, dendritic cells, and macrophages. Anti-vWFA2 autoantibodies located at the lamina densa bound to the dermal side of salt-split skin and induced blisters when transferred into healthy mice. Absence of CD8 T cells at time of immunization had no effect, whereas depletion of CD4 T cells during the same time period delayed autoantibody production and blisters. Collectively, we demonstrate the pathogenic relevance of Abs targeting the vWFA2 domain of COL7 and show the requirement of APC-induced CD4 T cells to induce experimental EBA.
在自身免疫性大疱性皮肤病 (AIBD) 中,自身抗体诱导皮肤或黏膜出现水疱,或两者皆有。利用 AIBD 动物模型,已充分阐明了持续产生自身抗体和水疱形成的机制。然而,导致最初产生自身抗体的机制尚未得到详细研究。获得性大疱性表皮松解症 (EBA) 是一种与 VII 型胶原 (COL7) 自身抗体相关的 AIBD。大多数 EBA 患者的血清识别非胶原域 1,包括血管性血友病因子 A 样域 2 (vWFA2)。通过用 GST-COL7 免疫诱导的实验性 EBA 中,疾病表现取决于遗传背景、Th1 极化和 GST 标签。在该模型中,裸鼠既不产生自身抗体也不产生水疱。EBA 诱导所需的 APC 和 T 细胞亚群仍不确定。我们通过用与内含肽融合的 vWFA2 免疫建立了一种新型 EBA 模型(缺少 GST 标签)。所有测试的小鼠品系均产生自身抗体,但水疱仅在携带 H2s 的小鼠中观察到。在免疫的小鼠中,检测到针对 vWFA2 的 CD4 T 细胞,其诱导需要 B 细胞、树突状细胞和巨噬细胞的存在。位于板层致密部的抗 vWFA2 自身抗体结合在盐裂皮肤的真皮侧,当转移到健康小鼠中时会引起水疱。免疫时缺乏 CD8 T 细胞没有影响,而在此期间耗尽 CD4 T 细胞会延迟自身抗体的产生和水疱的形成。总之,我们证明了靶向 COL7 的 vWFA2 结构域的 Abs 的致病性相关性,并显示 APC 诱导的 CD4 T 细胞在诱导实验性 EBA 中的必要性。