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T cells recognizing a peptide contaminant undetectable by mass spectrometry.
Brezar, Vedran; Culina, Slobodan; Østerbye, Thomas; Guillonneau, François; Chiappetta, Giovanni; Verdier, Yann; Vinh, Joelle; Wong, F Susan; Buus, Søren; Mallone, Roberto.
Afiliação
  • Brezar V; INSERM, U986, DeAR Lab Avenir, Saint Vincent de Paul Hospital, Paris, France.
PLoS One ; 6(12): e28866, 2011.
Article em En | MEDLINE | ID: mdl-22194932
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 ß-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 IGRP(206-214) using a novel method confirmed the identity of the contaminant, further underlining the immunodominance of IGRP(206-214). If left undetected, minute impurities in synthetic peptide preparations may thus give spurious results.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Peptídeos / Espectrometria de Massas / Linfócitos T CD8-Positivos Tipo de estudo: Prognostic_studies Limite: Animals / Humans Idioma: En Revista: PLoS One Ano de publicação: 2011 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Peptídeos / Espectrometria de Massas / Linfócitos T CD8-Positivos Tipo de estudo: Prognostic_studies Limite: Animals / Humans Idioma: En Revista: PLoS One Ano de publicação: 2011 Tipo de documento: Article