Pathogenic anti-desmoglein 3 mAbs cloned from a paraneoplastic pemphigus patient by phage display

Marwah A. Saleh, Ken Ishii, Jun Yamagami, Yuji Shirakata, Koji Hashimoto, Masayuki Amagai

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease associated with lymphoproliferative neoplasms and characterized by antibodies against plakins and desmoglein 3 (Dsg3). Anti-Dsg3 antibodies have a primary role in blister formation in PNP. In this study, we used phage display to clone monoclonal anti-Dsg3 antibodies from a PNP patient to further characterize their pathogenicity. We isolated 20 unique Dsg3-reactive mAbs, which we classified into four groups according to the heavy-chain complementarity-determining region 3 (CDR3) region. Genetic analyses demonstrated that three antibody groups used the VH1-46 gene (18 clones) and one group used the VH1-02 gene (2 clones). The results of an in vitro keratinocyte dissociation assay and a human skin organ culture injection assay showed that three antibodies displayed pathogenic activity in blister formation with different potencies. Epitope mapping using domain-swapped Dsg3/Dsg2 showed that these pathogenic mAbs bound Ca 2+ -dependent conformational epitopes in the middle portion of the extracellular region of Dsg3 (EC2 and EC3 domains), in contrast to most previously characterized pathogenic pemphigus vulgaris antibodies, which bound to the EC1 domain of Dsg3. These mAbs reflect the unique polyclonal nature of anti-Dsg3 antibodies in PNP and represent an important tool for detailing the pathophysiological mechanisms of blister formation in PNP.

Original languageEnglish
Pages (from-to)1141-1148
Number of pages8
JournalJournal of Investigative Dermatology
Volume132
Issue number4
DOIs
Publication statusPublished - 2012 Apr

Fingerprint

Desmoglein 3
Bacteriophages
Pemphigus
Display devices
Antibodies
Blister
Clone Cells
Epitopes
Assays
Plakins
Genes
Complementarity Determining Regions
Neoplasm Antibodies
Epitope Mapping
Organ Culture Techniques
Keratinocytes
Autoimmune Diseases
Virulence
Skin

ASJC Scopus subject areas

  • Dermatology
  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Pathogenic anti-desmoglein 3 mAbs cloned from a paraneoplastic pemphigus patient by phage display. / Saleh, Marwah A.; Ishii, Ken; Yamagami, Jun; Shirakata, Yuji; Hashimoto, Koji; Amagai, Masayuki.

In: Journal of Investigative Dermatology, Vol. 132, No. 4, 04.2012, p. 1141-1148.

Research output: Contribution to journalArticle

Saleh, Marwah A. ; Ishii, Ken ; Yamagami, Jun ; Shirakata, Yuji ; Hashimoto, Koji ; Amagai, Masayuki. / Pathogenic anti-desmoglein 3 mAbs cloned from a paraneoplastic pemphigus patient by phage display. In: Journal of Investigative Dermatology. 2012 ; Vol. 132, No. 4. pp. 1141-1148.
@article{247c20da8d0146da99a3f69c33db82d9,
title = "Pathogenic anti-desmoglein 3 mAbs cloned from a paraneoplastic pemphigus patient by phage display",
abstract = "Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease associated with lymphoproliferative neoplasms and characterized by antibodies against plakins and desmoglein 3 (Dsg3). Anti-Dsg3 antibodies have a primary role in blister formation in PNP. In this study, we used phage display to clone monoclonal anti-Dsg3 antibodies from a PNP patient to further characterize their pathogenicity. We isolated 20 unique Dsg3-reactive mAbs, which we classified into four groups according to the heavy-chain complementarity-determining region 3 (CDR3) region. Genetic analyses demonstrated that three antibody groups used the VH1-46 gene (18 clones) and one group used the VH1-02 gene (2 clones). The results of an in vitro keratinocyte dissociation assay and a human skin organ culture injection assay showed that three antibodies displayed pathogenic activity in blister formation with different potencies. Epitope mapping using domain-swapped Dsg3/Dsg2 showed that these pathogenic mAbs bound Ca 2+ -dependent conformational epitopes in the middle portion of the extracellular region of Dsg3 (EC2 and EC3 domains), in contrast to most previously characterized pathogenic pemphigus vulgaris antibodies, which bound to the EC1 domain of Dsg3. These mAbs reflect the unique polyclonal nature of anti-Dsg3 antibodies in PNP and represent an important tool for detailing the pathophysiological mechanisms of blister formation in PNP.",
author = "Saleh, {Marwah A.} and Ken Ishii and Jun Yamagami and Yuji Shirakata and Koji Hashimoto and Masayuki Amagai",
year = "2012",
month = "4",
doi = "10.1038/jid.2011.449",
language = "English",
volume = "132",
pages = "1141--1148",
journal = "Journal of Investigative Dermatology",
issn = "0022-202X",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Pathogenic anti-desmoglein 3 mAbs cloned from a paraneoplastic pemphigus patient by phage display

AU - Saleh, Marwah A.

AU - Ishii, Ken

AU - Yamagami, Jun

AU - Shirakata, Yuji

AU - Hashimoto, Koji

AU - Amagai, Masayuki

PY - 2012/4

Y1 - 2012/4

N2 - Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease associated with lymphoproliferative neoplasms and characterized by antibodies against plakins and desmoglein 3 (Dsg3). Anti-Dsg3 antibodies have a primary role in blister formation in PNP. In this study, we used phage display to clone monoclonal anti-Dsg3 antibodies from a PNP patient to further characterize their pathogenicity. We isolated 20 unique Dsg3-reactive mAbs, which we classified into four groups according to the heavy-chain complementarity-determining region 3 (CDR3) region. Genetic analyses demonstrated that three antibody groups used the VH1-46 gene (18 clones) and one group used the VH1-02 gene (2 clones). The results of an in vitro keratinocyte dissociation assay and a human skin organ culture injection assay showed that three antibodies displayed pathogenic activity in blister formation with different potencies. Epitope mapping using domain-swapped Dsg3/Dsg2 showed that these pathogenic mAbs bound Ca 2+ -dependent conformational epitopes in the middle portion of the extracellular region of Dsg3 (EC2 and EC3 domains), in contrast to most previously characterized pathogenic pemphigus vulgaris antibodies, which bound to the EC1 domain of Dsg3. These mAbs reflect the unique polyclonal nature of anti-Dsg3 antibodies in PNP and represent an important tool for detailing the pathophysiological mechanisms of blister formation in PNP.

AB - Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease associated with lymphoproliferative neoplasms and characterized by antibodies against plakins and desmoglein 3 (Dsg3). Anti-Dsg3 antibodies have a primary role in blister formation in PNP. In this study, we used phage display to clone monoclonal anti-Dsg3 antibodies from a PNP patient to further characterize their pathogenicity. We isolated 20 unique Dsg3-reactive mAbs, which we classified into four groups according to the heavy-chain complementarity-determining region 3 (CDR3) region. Genetic analyses demonstrated that three antibody groups used the VH1-46 gene (18 clones) and one group used the VH1-02 gene (2 clones). The results of an in vitro keratinocyte dissociation assay and a human skin organ culture injection assay showed that three antibodies displayed pathogenic activity in blister formation with different potencies. Epitope mapping using domain-swapped Dsg3/Dsg2 showed that these pathogenic mAbs bound Ca 2+ -dependent conformational epitopes in the middle portion of the extracellular region of Dsg3 (EC2 and EC3 domains), in contrast to most previously characterized pathogenic pemphigus vulgaris antibodies, which bound to the EC1 domain of Dsg3. These mAbs reflect the unique polyclonal nature of anti-Dsg3 antibodies in PNP and represent an important tool for detailing the pathophysiological mechanisms of blister formation in PNP.

UR - http://www.scopus.com/inward/record.url?scp=84858285073&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84858285073&partnerID=8YFLogxK

U2 - 10.1038/jid.2011.449

DO - 10.1038/jid.2011.449

M3 - Article

VL - 132

SP - 1141

EP - 1148

JO - Journal of Investigative Dermatology

JF - Journal of Investigative Dermatology

SN - 0022-202X

IS - 4

ER -