Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display

Aimee S. Payne, Ken Ishii, Stephen Kacir, Chenyan Lin, Hong Li, Yasushi Hanakawa, Kazuyuki Tsunoda, Masayuki Amagai, John R. Stanley, Don L. Siegel

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)888-899
Number of pages12
JournalJournal of Clinical Investigation
Volume115
Issue number4
DOIs
Publication statusPublished - 2005 Apr

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Pemphigus
Autoantibodies
Bacteriophages
Desmoglein 3
Antibodies
Desmogleins
Single-Chain Antibodies
Blister
Keratinocytes
Virulence
Epitopes
Mucous Membrane
Enzyme-Linked Immunosorbent Assay
Light
Skin
Injections
Serum
Genes
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display. / Payne, Aimee S.; Ishii, Ken; Kacir, Stephen; Lin, Chenyan; Li, Hong; Hanakawa, Yasushi; Tsunoda, Kazuyuki; Amagai, Masayuki; Stanley, John R.; Siegel, Don L.

In: Journal of Clinical Investigation, Vol. 115, No. 4, 04.2005, p. 888-899.

Research output: Contribution to journalArticle

Payne, Aimee S. ; Ishii, Ken ; Kacir, Stephen ; Lin, Chenyan ; Li, Hong ; Hanakawa, Yasushi ; Tsunoda, Kazuyuki ; Amagai, Masayuki ; Stanley, John R. ; Siegel, Don L. / Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display. In: Journal of Clinical Investigation. 2005 ; Vol. 115, No. 4. pp. 888-899.
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