Synergistic pathogenic effects of combined mouse monoclonal anti-desmoglein 3 IgG antibodies on pemphipus vulgaris blister formation
Synergistic pathogenic effects of combined mouse monoclonal anti-desmoglein 3 IgG antibodies on pemphipus vulgaris blister formation
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DOI：
10.1038/sj.jid.5700450
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发表时间：
2006-12-01
影响因子：
6.5
通讯作者：
Amagai, Masayuki
中科院分区：
文献类型：
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作者：
Kawasaki, Hiroshi;Tsunoda, Kazuyuki;Amagai, Masayuki
寻常型天疱疮 (PV) 是一种由抗桥粒芯糖蛋白 3 (Dsg3) IgG 抗体引起的自身免疫性水疱性疾病。此前，我们通过过继转移免疫小鼠或幼稚 Dsg3(-/-) 小鼠的脾细胞，生成了活跃的 PV 小鼠模型。在本研究中，我们从通过转移初始 Dsg3(-/-) 脾细胞产生的 PV 模型小鼠中分离出 10 种抗 Dsg3 IgG mAb（NAK 系列）。我们使用结构域交换和点突变的 Dsg1/Dsg3 分子表征了它们的表位，并在三种不同的测定中检查了它们在水疱形成中的致病活性。在使用新生小鼠的被动转移模型中，当与落叶型天疱疮 (PF) 患者的抗 Dsg1 IgG 自身抗体最低致病剂量的一半一起注射时，10 种 NAK mAb 中的 8 种表现出致病活性。当通过腹膜注射将单个杂交瘤克隆接种到成年Rag2(-/-)小鼠中时，没有一种单克隆抗体能够诱导PV表型。 NAK mAb 在使用原代培养的小鼠角质形成细胞的体外解离测定中显示出一系列效力。有趣的是，当将识别不同表位的多个杂交瘤克隆接种到组合受体小鼠中时，小鼠出现了 PV 表型。体外解离测定证实组合的 NAK mAb 具有协同致病作用。这些发现表明，虽然单个抗 Dsg3 IgG 不足以引起成年小鼠出现水疱，但多个抗 Dsg3 IgG 一起可以诱导 PV 表型。这些单克隆抗体将提供一个有价值的工具来研究水疱形成的分子机制，模仿患者体内发现的多克隆 IgG 抗体的作用。
Pemphigus vulgaris (PV) is an autoimmune blistering disease caused by anti-desmoglein 3 (Dsg3) IgG antibodies. Previously, we generated an active mouse model for PV by adoptive transfer of splenocytes from immunized or naive Dsg3(-/-) mice. In this study, we isolated 10 anti-Dsg3 IgG mAbs (NAK-series) from PV model mice generated by transfer of naive Dsg3(-/-) splenocytes. We characterized their epitopes using domain-swapped and point-mutated Dsg1/Dsg3 molecules and examined their pathogenic activities in blister formation in three different assays. In a passive transfer model using neonatal mice, eight of 10 NAK mAbs showed pathogenic activity when injected together with half the minimum pathogenic dose of anti-Dsg1 IgG autoantibodies from pemphigus foliaceus (PF) patients. None of the mAbs could induce the PV phenotype when individual hybridoma clones were inoculated by peritoneal injection into adult Rag2(-/-) mice. NAK mAbs displayed a range of potency in an in vitro dissociation assay using primary cultured mouse keratinocytes. Interestingly, when multiple hybridoma clones recognizing different epitopes were inoculated in combination recipient mice developed the PV phenotype. In vitro dissociation assays confirmed that combined NAK mAbs had synergistic pathogenic effects. These findings indicate that although an individual anti-Dsg3 IgG is not sufficient to cause blistering in adult mice, several together can induce the PV phenotype. These mAbs will provide a valuable tool to investigate the molecular mechanisms of blister formation, mimicking the effects of the polyclonal IgG antibodies found in patients.