Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display
Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display
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DOI：
10.1172/jci200524185
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发表时间：
2005-04-01
影响因子：
15.9
通讯作者：
Siege, DL
中科院分区：
文献类型：
--
作者：
Payne, AS;Ishii, K;Siege, DL
天疱疮是一种危及生命的皮肤和粘膜水疱性疾病，由桥粒粘附蛋白桥粒糖蛋白 3 (Dsg3) 和 Dsg1 的致病性自身抗体引起。使用多克隆患者血清很难表征抗体致病性的机制。使用抗体噬菌体展示，我们从患有活动性皮肤粘膜寻常型天疱疮的患者中分离出人类抗 Dsg mAb 的单链可变区片段 (scFv)。 ScFv mAb 表现出单独与 Dsg3 或 Dsg1 结合，或与 Dsg3 和 Dsg1 两者结合。抑制 ELISA 显示，这些 scFv 定义的表位被来自多个天疱疮患者的自身抗体阻断。将 scFv 注射到新生小鼠体内，发现了 2 种致病性 scFv，它们引起的水泡在组织学上与在天疱疮患者中观察到的情况相似。类似地，这 2 个 scFv（而非其他）诱导培养的人角质形成细胞的细胞片解离，表明致病性和非致病性抗体均被分离。这些 mAb 的遗传分析显示重链和轻链基因使用的限制模式，这对于具有不同桥粒芯糖蛋白结合特异性的 scFv 来说是不同的。这些天疱疮单克隆抗体的详细表征应该有助于更好地了解疾病的免疫发病机制，并制定更有针对性的治疗方法。
Pemphigus is a life-threatening blistering disorder of the skin and mucous membranes caused by pathogenic autoantibodies to desmosomal adhesion proteins desmoglein 3 (Dsg3) and Dsg1. Mechanisms of antibody pathogenicity are difficult to characterize using polyclonal patient sera. Using antibody phage display, we have isolated repertoires of human anti-Dsg mAbs as single-chain variable-region fragments (scFvs) from a patient with active mucocutaneous pemphigus vulgaris. ScFv mAbs demonstrated binding to Dsg3 or Dsg1 alone, or both Dsg3 and Dsg1. Inhibition ELISA showed that the epitopes defined by these scFvs are blocked by autoantibodies from multiple pemphigus patients. Injection of scFvs into neonatal mice identified 2 pathogenic scFvs that caused blisters histologically similar to those observed in pemphigus patients. Similarly, these 2 scFvs, but not others, induced cell sheet dissociation of cultured human keratinocytes, indicating that both pathogenic and nonpathogenic antibodies were isolated. Genetic analysis of these mAbs showed restricted patterns of heavy and light chain gene usage, which were distinct for scFvs with different desmoglein-binding specificities. Detailed characterization of these pemphigus mAbs should lead to a better understanding of the immunopathogenesis of disease and to more specifically targeted therapeutic approaches.