title: Human T-cells recognise N-terminally Fmoc-modified peptide.
abstract: We aimed to generate T-cell clones specific for human pre-proinsulin. An HLA DQ8, CD4+ T-cell clone that recognised a 10mer (C65-A9) peptide from pre-proinsulin was isolated. Further analysis revealed that the clone responded neither to recombinant proinsulin nor to re-synthesised C65-A9 peptide. Analysis of the original peptide revealed minor contamination (<0.5%) with an N-terminal Fmoc adduct. This peptide was synthesised and shown to stimulate the clone. Thus, Fmoc-modified peptides, which are common contaminants in synthetic peptides, can stimulate human CD4+ T-cells. This finding has important implications for the use of synthetic peptides in screening and epitope mapping studies and their use as vaccines in humans.
authors.affiliation: Autoimmunity and Transplantation Division, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, 1G Royal Parade, Parkville, Vic. 3050, Australia. mannering@wehi.edu.au
authors.forename: Stuart I
authors.initials: SI
authors.lastname: Mannering
authors.forename: Anthony W
authors.initials: AW
authors.lastname: Purcell
authors.forename: Margo C
authors.initials: MC
authors.lastname: Honeyman
authors.forename: James
authors.initials: J
authors.lastname: McCluskey
authors.forename: Leonard C
authors.initials: LC
authors.lastname: Harrison
mesh_terms: D001323:Autoantibodies / Q000032:analysis; D002469:Cell Separation; D002851:Chromatography, High Pressure Liquid; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin; D013058:Mass Spectrometry; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D011384:Proinsulin / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D011498:Protein Precursors / Q000737:chemistry / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D013967:Thyroiditis, Autoimmune / Q000276:immunology
chemical_list: C482904:9-fluorenylmethoxycarbonyl-arginyl-glycyl-isoleucyl-valyl-glutamyl-glutaminyl-cysteinyl-cysteinyl-threonyl-serine; D001323:Autoantibodies; D006680:HLA Antigens; D007328:Insulin; D010446:Peptide Fragments; D011498:Protein Precursors; C013737:preproinsulin; D011384:Proinsulin