Characterization of desmoglein-3 epitope region peptides as synthetic antigens: analysis of theirin vitroT cell stimulating efficacy, cytotoxicity, stability, and their conformational features

Abstract

Oligopeptides, representing B-and/or T-cell epitope(s) are promising candidates as future synthetic antigens for studying human autoimmune disorders. Synthetic peptides could be as efficient in inducing in vitro T-cell responses as recombinant proteins. We have studied desmoglein 3 (Dsg3) peptides representing T-cell epitope regions to determine their capability to induce in vitro T-cell response and to analyze the influence of their solution conformation.The design of synthetic peptides used in this study was based on the previously described Dsg3 T-cell epitope regions and resulted in new N-terminally truncated versions. The peptides were prepared on solid phase and chemically characterized. In order to establish structureactivity relationship, the solution conformation of the synthetic peptides has been investigated using ECD spectroscopy. The in vitro T-cell stimulating efficacy as well as in vitro cytotoxicity and stability of the peptides have been determined on peripheral blood mononuclear cells (PBMC) isolated from whole blood of Pemphigus vulgaris (PV) patients and also from healthy donors. After 20 hours stimulation the IFN-γ content of the supernatants was measured by ELISA. Our data suggested that the in vitro IFN-γ production of PBMC culture derived from healthy donors and PV patients induced by synthetic peptides as antigens are markedly different. Comparative analysis of in vitro activity and the capability of oligopeptides to form ordered or unordered secondary structure was performed. We measured different in vitro T-cell stimulating activity on PBMC derived from patients and healthy donors, especially with synthetic peptides bearing high solvent sensibility which might suggest backbone flexibility.

Biokonjugátumok – összefogni érdemes

“…mucin glikoproteinek) illetve az autoimmun betegségekben autoantigénként szerepet játszó fehérjék (pl. filaggrin, fibrin, desmoglein 14,15 ) a poszt-transzlációs módosulás elõtt nem viselkednek "idegen" fehérjeként, míg azt követõen immunreakciót váltanak ki. …”