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/jci24185
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发表时间：
2005-04
期刊：
影响因子：
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通讯作者：
A. Payne;K. Ishii;Stephen Kacir;Chenyan Lin;Hong Li;Y. Hanakawa;K. Tsunoda;M. Amagai;J. Stanley;D. Siegel
中科院分区：
文献类型：
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作者：
A. Payne;K. Ishii;Stephen Kacir;Chenyan Lin;Hong Li;Y. Hanakawa;K. Tsunoda;M. Amagai;J. Stanley;D. Siegel
天疱疮是一种威胁生命的皮肤和粘膜起泡性疾病，由抗桥粒黏附蛋白3(Dsg3)和DSG1的病理性自身抗体引起。抗体致病的机制很难用多克隆患者血清来表征。利用抗体噬菌体展示技术，我们从1例活动期寻常型黏膜天疱疮患者体内分离出人源抗DSG单抗作为单链可变区片段(ScFv)。单链抗体仅与Dsg3或DSG1结合，或同时与Dsg3和DSG1结合。抑制酶联免疫吸附试验显示，这些单链抗体所定义的表位被来自多个天疱疮患者的自身抗体所阻断。将单链抗体注射到新生小鼠中，发现了两种引起水泡的病原性单链抗体，组织学上与在天疱疮患者中观察到的相似。同样，这两种单链抗体可诱导培养的人角质形成细胞膜解离，说明致病抗体和非致病抗体均被分离出来。这些单抗的遗传分析显示出重链和轻链基因使用的限制性模式，这对于具有不同桥粒粘连蛋白结合特异性的单链抗体是不同的。这些天疱疮单抗的详细特征应该有助于更好地了解疾病的免疫发病机制，并找到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.