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 ORCID: https://orcid.org/0000-0002-2812-8845, 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:	Schools > Medicine Research Institutes & Centres > 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:	18 May 2023 14:46
URI:	https://orca.cardiff.ac.uk/id/eprint/21925

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