Detection of low avidity desmoglein 3-reactive T cells in pemphigus vulgaris using HLA-DR beta 10402 tetramers

• Published in: Clinical immunology (Orlando, Fla.) Vol. 122; no. 3; pp. 330 - 337
• Main Authors: Veldman, Christian, Eming, Rüdiger, Wolff-Franke, Sonja, Sonderstrup, Grete, Kwok, William W, Hertl, Michael
• Format: Journal Article
• Language: English
• Published: United States 01.03.2007
• Subjects: Alleles; Amino Acid Sequence; Animals; Cell Adhesion - immunology; Cells, Cultured; Desmoglein 3 - immunology; Desmoglein 3 - metabolism; HLA-DR Antigens - immunology; HLA-DR Antigens - metabolism; HLA-DRB1 Chains; Humans; Hybridomas; Mice; Mice, Transgenic; Molecular Sequence Data; Pemphigus - immunology; Pemphigus - pathology; Protein Binding - immunology; T-Lymphocyte Subsets - immunology; T-Lymphocyte Subsets - metabolism
• ISSN: 1521-6616
• DOI: 10.1016/j.clim.2006.09.014

In the present study, we developed a HLA class II tetramer-based detection system utilizing DRB1*0402 tetramers loaded with recently identified immunodominant peptides of desmoglein 3 (Dsg3), the major autoantigen of pemphigus vulgaris (PV). Initial experiments demonstrated staining of a Dsg3-reactive T cell hybridoma which was derived from HLA-DR0402-transgenic mice with loaded PE-labeled DRbeta1*0402 tetramers. However, staining of autoreactive T cell clones (TCC) derived from PV patients resulted only in positive staining by addition of exogenous peptides to the staining reactions. There was a dose-dependent specific binding of TCC to the tetramers with the agonistic Dsg3 peptide which was not altered by exogenous unrelated Dsg3 peptide. Noteworthy, the TCC did not stain with HLA-DR4 tetramers complexed with unrelated Dsg3 peptides. The findings of this study suggest that HLA class II tetramers may provide a highly specific approach to monitor ex vivo the T cellular autoimmune response against Dsg3 in patients with PV.