Development of chimeric molecules for recognition and targeting of antigen-specific B cells in pemphigus vulgaris

C. M. Proby, T. Ota, H. Suzuki, S. Koyasu, S. Gamou, N. Shimizu, J. K. Wahl, M. J. Wheelock, T. Nishikawa, Masayuki Amagai

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Abstract

Pemphigus vulgaris (PV) is an autoimmune blistering disease characterized by circulating pathogenic IgG antibodies against desmoglein 3 (Dsg3). The purpose of this study was to develop chimeric molecules for specific recognition and elimination of autoimmune B cells in PV. Mouse hybridoma cell lines producing anti-Dsg3 antibody (SH1O, 12A2) were developed as an in vitro model system for targeting B cells. Dsg3-GFP, a baculoprotein containing the entire extracellular domain of Dsg3 fused with green fluorescence protein, recognized and targeted the hybridoma cells through their surface immunoglobulin receptors in an antigen-specific way. The epitopes of these monoclonal antibodies were mapped on the amino terminal EC1 and part of EC2, a region considered functionally important in cadherins. Chimeric toxin molecules containing the mapped region (Dsg3ΔN1) and modified Pseudomonas exotoxin were produced in bacteria (Dsg3ΔN1-PE40-KDEL, PE37- Dsg3ΔN1-KDEL) and tested in vitro on hybridoma cell lines. The chimeric toxins, but not Dsg3ΔN1 alone, showed dose-dependent toxic activity with a reduction in hybridoma cell number to 40-60% of toxin-negative control cultures, compared with little or no effect on anti-Dsg3-negative hybridoma cells. Furthermore, these toxins showed toxic effects on anti-Dsg3 IgG- producing B cells from Dsg3ΔN1-immunized mice, with a 60% reduction in cell number compared with Dsg3ΔN1 alone. Thus, specific recognition and targeting of antigen-specific B cells in PV was demonstrated; this strategy may hold promise as a future therapeutic option for PV and other autoimmune diseases.

Original languageEnglish (US)
Pages (from-to)321-330
Number of pages10
JournalBritish Journal of Dermatology
Volume142
Issue number2
DOIs
StatePublished - Mar 8 2000

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Keywords

  • Autoimmune disease
  • Cadherin
  • Desmoglein
  • Recombinant toxin

ASJC Scopus subject areas

  • Dermatology

Cite this

Proby, C. M., Ota, T., Suzuki, H., Koyasu, S., Gamou, S., Shimizu, N., Wahl, J. K., Wheelock, M. J., Nishikawa, T., & Amagai, M. (2000). Development of chimeric molecules for recognition and targeting of antigen-specific B cells in pemphigus vulgaris. British Journal of Dermatology, 142(2), 321-330. https://doi.org/10.1046/j.1365-2133.2000.03328.x