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
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Research Support, Non-U.S. Gov't
MeSH terms
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Autoantibodies / analysis
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Cell Separation
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Chromatography, High Pressure Liquid
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Clone Cells
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Enzyme-Linked Immunosorbent Assay
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Epitope Mapping
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HLA Antigens / immunology
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Humans
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In Vitro Techniques
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Insulin
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Mass Spectrometry
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Peptide Fragments / chemistry
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Peptide Fragments / isolation & purification*
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Proinsulin / chemistry
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Proinsulin / immunology*
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Proinsulin / isolation & purification*
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Protein Precursors / chemistry
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Protein Precursors / immunology*
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T-Lymphocytes / immunology*
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Thyroiditis, Autoimmune / immunology
Substances
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9-fluorenylmethoxycarbonyl-arginyl-glycyl-isoleucyl-valyl-glutamyl-glutaminyl-cysteinyl-cysteinyl-threonyl-serine
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Autoantibodies
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HLA Antigens
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Insulin
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Peptide Fragments
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Protein Precursors
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preproinsulin
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Proinsulin