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T cells recognizing a peptide contaminant undetectable by mass spectrometry

Oresic, Matej, Brezar, Vedran, Culina, Slobodan, Østerbye, Thomas, Guillonneau, François, Chiappetta, Giovanni, Verdier, Yann, Vinh, Joelle, Wong, Florence Susan, Buus, Søren and Mallone, Roberto 2011. T cells recognizing a peptide contaminant undetectable by mass spectrometry. PLoS ONE 6 (12) , e28866. 10.1371/journal.pone.0028866

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Abstract

Synthetic peptides are widely used in immunological research as epitopes to stimulate their cognate T cells. These preparations are never completely pure, but trace contaminants are commonly revealed by mass spectrometry quality controls. In an effort to characterize novel major histocompatibility complex (MHC) Class I-restricted β-cell epitopes in non-obese diabetic (NOD) mice, we identified islet-infiltrating CD8+ T cells recognizing a contaminating peptide. The amount of this contaminant was so small to be undetectable by direct mass spectrometry. Only after concentration by liquid chromatography, we observed a mass peak corresponding to an immunodominant islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)206-214 epitope described in the literature. Generation of CD8+ T-cell clones recognizing IGRP206-214 using a novel method confirmed the identity of the contaminant, further underlining the immunodominance of IGRP206-214. If left undetected, minute impurities in synthetic peptide preparations may thus give spurious results.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Subjects: R Medicine > R Medicine (General)
Additional Information: This research was supported by the Juvenile Diabetes Research Foundation (JDRF grant 1-2008-106), the European Foundation for the Study of Diabetes (EFSD/JDRF/Novo Nordisk European Programme in Type 1 Diabetes Research 2007) and the Domaine d'Intérêt Majeur: Cardiovasculaire – Obésité – Diabète (Ile-de-France CODDIM). R.M. is an INSERM Avenir Investigator. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher: PLoS
ISSN: 1932-6203
Last Modified: 04 Jun 2017 03:29
URI: http://orca.cf.ac.uk/id/eprint/21925

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