Department of Dermatology, Hokkaido University, Graduate School of Medicine, Sapporo, Japan.
J Immunol. 2010 Dec 15;185(12):7746-55. doi: 10.4049/jimmunol.1000667. Epub 2010 Nov 12.
Bullous pemphigoid (BP) is an autoimmune blistering disease caused by IgG autoantibodies targeting the noncollagenous 16A (NC16A) domain of human collagen 17 (hCOL17), which triggers blister formation via complement activation. Previous in vitro analysis demonstrated that IgG1 autoantibodies showed much stronger pathogenic activity than IgG4 autoantibodies; however, the exact pathogenic role of IgG1 autoantibodies has not been fully demonstrated in vivo. We constructed a recombinant IgG1 mAb against hCOL17 NC16A from BP patients. In COL17-humanized mice, this mAb effectively reproduced a BP phenotype that included subepidermal blisters, deposition of IgG1, C1q and C3, neutrophil infiltration, and mast cell degranulation. Subsequently, alanine substitutions at various C1q binding sites were separately introduced to the Fc region of the IgG1 mAb. Among these mutated mAbs, the one that was mutated at the P331 residue completely failed to activate the complement in vitro and drastically lost pathogenic activity in COL17-humanized mice. These findings indicate that P331 is a key residue required for complement activation and that IgG1-dependent complement activation is essential for blister formation in BP. This study is, to our knowledge, the first direct evidence that IgG1 Abs to hCOL17 NC16A can induce blister formation in vivo, and it raises the possibility that IgG1 mAbs with Fc modification may be used to block pathogenic epitopes in autoimmune diseases.
大疱性类天疱疮(BP)是一种由 IgG 自身抗体引起的自身免疫性水疱病,该抗体针对人类胶原蛋白 17(hCOL17)的非胶原 16A(NC16A)结构域,通过补体激活引发水疱形成。先前的体外分析表明,IgG1 自身抗体比 IgG4 自身抗体具有更强的致病性;然而,IgG1 自身抗体在体内的确切致病作用尚未完全证实。我们从 BP 患者中构建了针对 hCOL17 NC16A 的重组 IgG1 mAb。在 COL17 人源化小鼠中,该 mAb 有效地复制了 BP 表型,包括表皮下水疱、IgG1、C1q 和 C3 的沉积、中性粒细胞浸润和肥大细胞脱颗粒。随后,在 IgG1 mAb 的 Fc 区域分别引入了各种 C1q 结合位点的丙氨酸取代。在这些突变 mAb 中,P331 残基发生突变的 mAb 完全不能在体外激活补体,并且在 COL17 人源化小鼠中丧失了明显的致病活性。这些发现表明 P331 是补体激活所必需的关键残基,并且 IgG1 依赖性补体激活对于 BP 中的水疱形成是必不可少的。就我们所知,这项研究首次提供了直接证据,表明 hCOL17 NC16A 的 IgG1 Abs 可在体内诱导水疱形成,并提出了对 Fc 进行修饰的 IgG1 mAb 可能用于阻断自身免疫性疾病中致病性表位的可能性。