Human T-cells recognise N-terminally Fmoc-modified peptide

Vaccine. 2003 Sep 8;21(25-26):3638-46. doi: 10.1016/s0264-410x(03)00402-x.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantibodies / analysis
  • Cell Separation
  • Chromatography, High Pressure Liquid
  • Clone Cells
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping
  • HLA Antigens / immunology
  • Humans
  • In Vitro Techniques
  • Insulin
  • Mass Spectrometry
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification*
  • Proinsulin / chemistry
  • Proinsulin / immunology*
  • Proinsulin / isolation & purification*
  • Protein Precursors / chemistry
  • Protein Precursors / immunology*
  • T-Lymphocytes / immunology*
  • Thyroiditis, Autoimmune / immunology

Substances

  • 9-fluorenylmethoxycarbonyl-arginyl-glycyl-isoleucyl-valyl-glutamyl-glutaminyl-cysteinyl-cysteinyl-threonyl-serine
  • Autoantibodies
  • HLA Antigens
  • Insulin
  • Peptide Fragments
  • Protein Precursors
  • preproinsulin
  • Proinsulin