Functional analysis of two tetanus toxin universal T cell epitopes in their interaction with DR1101 and DR1104 alleles.
In this study we have investigated the interaction between two DR11 alleles (DB*1101 and DB*1104) and two previously described tetanus toxin (tt) universal T cell epitopes P2(tt830-843) and P30(tt947-967) by means of a functional cytotoxic competition assay. Both truncation analysis and single alanine substitution analysis were performed. In addition, the capacity of truncated and single alanine substituted peptides to be recognized by human T cell clones from donors bearing the DR1101 or DR1104 alleles was assessed. In the case of truncated peptides the same binding and recognition pattern was observed with both alleles. Longer peptides were better competitors and more potent stimulators, a result that should be taken into account when these peptides are used as immunogens. None of the single alanine substitutions could abrogate or strongly diminish the inhibitory capacity of the analogues tested indicating the lack of strong "anchor residues" present in P2 and P30 and implicated in DR binding. In addition, although the original peptide sequences were presented to specific T cell clones with comparable efficiency, some of the alanine single substituted peptides were better recognized in association with one of the alleles by clones derived from individuals bearing the homologous allele. The only exception was the tt951-967 analogue ttW955A, which was preferentially recognized in association with the DR1104 allele regardless of the clone tested. This suggests that, although it binds to both alleles with comparable efficiency, the MHC-peptide complex so formed is conformationally distinguishable by specific T cell clones.
Valmori
D
D
Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland.
Sabbatini
A
A
Lanzavecchia
A
A
Corradin
G
G
Matricardi
P M
PM
D000483:Alleles; D000595:Amino Acid Sequence; D000939:Epitopes; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D059810:HLA-DR Serological Subtypes; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000276:immunology*; D013744:Tetanus Toxin / Q000276:immunology*
D000939:Epitopes; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; C080766:HLA-DR11 antigen; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D013744:Tetanus Toxin