Department of Dermatology, Ruprecht-Karls University Heidelberg, Heidelberg, Germany.
Department of Dermatology, University of Lübeck, Lübeck, Germany.
J Allergy Clin Immunol. 2018 Dec;142(6):1831-1842.e7. doi: 10.1016/j.jaci.2018.04.006. Epub 2018 Apr 26.
Autoimmune bullous diseases/dermatoses (AIBDs) are severe autoantibody-mediated skin diseases. The pathogenic relevance of autoreactive CD4 T cells for the induction of autoantibody production remains to be fully evaluated. Scurfy mice lack functional regulatory T (Treg) cells, experience spontaneous activation of autoreactive CD4 T cells, and display severe erosive skin lesions suggestive of AIBDs.
We sought to determine whether AIBDs develop in Treg cell-deficient scurfy mice.
Histology, indirect immunofluorescence (IF) microscopy, direct IF, and ELISA were used to prove the presence of AIBDs in scurfy mice. Monoclonal autoantibodies from sera of scurfy mice were screened by using indirect IF on murine skin, and immunoprecipitation and mass spectrometry were used for target antigen identification, followed by confirmation in modified human embryonic kidney cells and murine keratinocytes. Pathogenicity was determined by injecting the autoantibody into neonatal mice and transferring scurfy CD4 T cells into nu/nu mice.
Autoantibodies against different known autoantigens of AIBDs spontaneously develop in scurfy mice. Histology reveals subepidermal blisters, and direct IF of skin of scurfy mice shows a predominant linear staining pattern. The mAb 20B12 shows a linear staining pattern in indirect IF, recognizes the murine hemidesmosomal protein bullous pemphigoid antigen 230 (BP230) as the target antigen, and cross-reacts with human BP230. Purified mAb 20B12 induces subepidermal blisters in neonatal mice. Transfer of scurfy CD4 T cells is sufficient to induce antibodies with reactivity to AIBD autoantigens and subepidermal blisters in the skin of recipient T cell-deficient nu/nu mice.
We show that the absence of Treg cells leads to AIBDs by pathogenic autoantibodies targeting BP230.
自身免疫性大疱性疾病/皮肤病（AIBD）是严重的自身抗体介导的皮肤疾病。自身反应性 CD4 T 细胞在诱导自身抗体产生中的发病相关性仍有待充分评估。Scurfy 小鼠缺乏功能性调节性 T（Treg）细胞，会自发激活自身反应性 CD4 T 细胞，并表现出严重的侵蚀性皮肤损伤，提示 AIBD 的发生。
我们旨在确定 Treg 细胞缺陷的 Scurfy 小鼠是否会发生 AIBD。
采用组织学、间接免疫荧光（IF）显微镜、直接 IF 和 ELISA 来证实 Scurfy 小鼠存在 AIBD。使用间接 IF 在鼠皮上筛选来自 Scurfy 小鼠血清中的单克隆自身抗体，通过免疫沉淀和质谱分析进行靶抗原鉴定，随后在改良的人胚肾细胞和鼠角质细胞中进行确认。通过将自身抗体注射到新生小鼠中并将 Scurfy CD4 T 细胞转移到 nu/nu 小鼠中来确定致病性。
Scurfy 小鼠中自发产生针对不同已知 AIBD 自身抗原的自身抗体。组织学显示表皮下疱，Scurfy 小鼠皮肤的直接 IF 显示出主要的线性染色模式。单克隆抗体 20B12 在间接 IF 中显示线性染色模式，识别鼠半桥粒蛋白大疱性类天疱疮抗原 230（BP230）为靶抗原，并与人类 BP230 发生交叉反应。纯化的单克隆抗体 20B12 可诱导新生小鼠表皮下疱。Scurfy CD4 T 细胞的转移足以在受体 T 细胞缺陷型 nu/nu 小鼠的皮肤中诱导针对 AIBD 自身抗原和表皮下疱的抗体。
我们表明，Treg 细胞缺失会导致致病性自身抗体针对 BP230 而引发 AIBD。
