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, James K Wahl, M. J. Wheelock, T. Nishikawa, Masayuki Amagai

Research output: Contribution to journalArticle

31 Citations (Scopus)

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

Fingerprint

Desmoglein 3
Pemphigus
Hybridomas
B-Lymphocytes
Antigens
Immunotoxins
Poisons
Autoimmune Diseases
Cell Count
Immunoglobulin G
Cell Line
Exotoxins
B-Cell Antigen Receptors
Antigen Receptors
Antibodies
Cadherins
Pseudomonas
Epitopes
Fluorescence
Monoclonal Antibodies

Keywords

  • Autoimmune disease
  • Cadherin
  • Desmoglein
  • Recombinant toxin

ASJC Scopus subject areas

  • Dermatology

Cite this

Development of chimeric molecules for recognition and targeting of antigen-specific B cells in pemphigus vulgaris. / Proby, C. M.; Ota, T.; Suzuki, H.; Koyasu, S.; Gamou, S.; Shimizu, N.; Wahl, James K; Wheelock, M. J.; Nishikawa, T.; Amagai, Masayuki.

In: British Journal of Dermatology, Vol. 142, No. 2, 08.03.2000, p. 321-330.

Research output: Contribution to journalArticle

Proby, CM, Ota, T, Suzuki, H, Koyasu, S, Gamou, S, Shimizu, N, Wahl, JK, Wheelock, MJ, 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, vol. 142, no. 2, pp. 321-330. https://doi.org/10.1046/j.1365-2133.2000.03328.x
Proby, C. M. ; Ota, T. ; Suzuki, H. ; Koyasu, S. ; Gamou, S. ; Shimizu, N. ; Wahl, James K ; Wheelock, M. J. ; Nishikawa, T. ; Amagai, Masayuki. / Development of chimeric molecules for recognition and targeting of antigen-specific B cells in pemphigus vulgaris. In: British Journal of Dermatology. 2000 ; Vol. 142, No. 2. pp. 321-330.
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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.",
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AU - Shimizu, N.

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