Submolecular recognition regions of the HN domain of the heavy chain of botulinum neurotoxin type A by T cells from toxin-treated cervical dystonia patients

Abstract

We have recently reported the submolecular T-cell recognition profile of the C-terminal half (H_C, residues 855-1296) of the heavy (H) chain of botulinum neurotoxin type A (BoNT/A) with peripheral blood lymphocytes (PBL) from 25 BoNT-treated cervical dystonia (CD) patients. In the current study, we describe the mapping of the T-cell responses of the patients to the N-terminal half (H_N, residues 449-859) of the heavy chain by using 29 synthetic overlapping peptides encompassing the entire H_N domain of BoNT/A. The profiles of the T-cell responses to the peptides varied among the patients. Samples from 14 patients treated solely with BoNT/A recognized 1-9 (average 3.7) peptides/sample at Z>3.0 level. Three peptide regions representing residues 631-649, 659-677 and 743-761 were frequently recognized by 29-64% of the patients. In patients with positive anti-BoNT/A antibody responses the overall positive T cell responses to the H_N peptides were significantly increased compared to antibody-negative patients. Influence of treatment parameters on the T-cell recognition of the H_N peptides was also observed. The results were compared with those of previously identified H_C region.

Keywords: Botulinum neurotoxin type A; Cervical dystonia; H(N) domain; MHC control; Synthetic peptides; T-cell epitopes.