1. T cells recognizing a peptide contaminant undetectable by mass spectrometry
- Author
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Slobodan Culina, Vedran Brezar, Thomas Osterbye, Yann Verdier, Giovanni Chiappetta, F. Susan Wong, Joëlle Vinh, Roberto Mallone, Søren Buus, François Guillonneau, VINH, Joëlle, Immunologie et génétique du diabète de type 1, génétique multifactorielle en endocrinologie pédiatrique (U986), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratory of Experimental Immunology [Copenhagen], Department of Immunology and Microbiology [Copenhagen], Faculty of Health and Medical Sciences, University of Copenhagen = Københavns Universitet (UCPH)-University of Copenhagen = Københavns Universitet (UCPH)-Faculty of Health and Medical Sciences, University of Copenhagen = Københavns Universitet (UCPH)-University of Copenhagen = Københavns Universitet (UCPH), Plateforme protéomique 3P5 [Institut Cochin] (3P5), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Spectrométrie de Masse Biologique et Protéomique (USR3149 / FRE2032) (SMBP), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Centre for Endocrine and Diabetes Sciences, Cardiff University, This research was supported by the Juvenile Diabetes Research Foundation (JDRF grant 1-2008-106), the European Foundation for the Study ofDiabetes (EFSD/JDRF/Novo Nordisk European Programme in Type 1 Diabetes Research 2007) and the Domaine d’Inte´reˆt Majeur: Cardiovasculaire – Obe´site´ –Diabe`te (Ile-de-France CODDIM), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Descartes - Paris 5 (UPD5), Institut des Maladies Emergentes et des Thérapies Innovantes (IMETI), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, University of Copenhagen = Københavns Universitet (KU)-University of Copenhagen = Københavns Universitet (KU)-Faculty of Health and Medical Sciences, University of Copenhagen = Københavns Universitet (KU)-University of Copenhagen = Københavns Universitet (KU), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Verdier, Yann, Physiopathologie et immunothérapies dans l’infection VIH, GHU A. Chenevier - Henri Mondor (Inserm U955, équipe 16), Neurobiologie et diversité cellulaire (NDC), Benaroya Research Institute at Virginia Mason and Department of Immunology, University of Washington School of Medicine, Service de diabétologie [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], CHU Cochin [AP-HP], Université Paris Descartes - Paris 5 ( UPD5 ), Institut des Maladies Emergentes et des Thérapies Innovantes ( IMETI ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris-Saclay, Laboratory of Experimental Immunology, University of Copenhagen ( KU ), Plateforme protéomique 3P5 [Institut Cochin] ( 3P5 ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Unité de Spectrométrie de Masse Biologique et Protéomique, ESPCI ParisTech-Centre National de la Recherche Scientifique ( CNRS ), CNRS/ESPCI ParisTech USR 3149, Centre National de la Recherche Scientifique ( CNRS ), Department of Cellular and Molecular Medicine, University of Bristol, University Walk, Faculty of Health Sciences-University of Copenhagen ( KU ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre National de la Recherche Scientifique (CNRS)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)
- Subjects
Proteomics ,Mouse ,MESH: Mice, Inbred NOD ,MESH: Sequence Analysis, Protein ,Autoimmunity ,Peptide ,MESH: Amino Acid Sequence ,MESH : Glucose-6-Phosphatase ,CD8-Positive T-Lymphocytes ,Mass Spectrometry ,Epitope ,MESH : Proteins ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,Sequence Analysis, Protein ,MESH: Animals ,MESH: Proteins ,Immune Response ,Peptide sequence ,chemistry.chemical_classification ,0303 health sciences ,MESH : Peptides ,MESH: Peptides ,MESH : Amino Acid Sequence ,MESH: CD8-Positive T-Lymphocytes ,3. Good health ,MESH: Reproducibility of Results ,[SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology ,[ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,Glucose-6-Phosphatase ,Medicine ,endocrine system ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,Immune Cells ,Science ,Molecular Sequence Data ,Immunology ,Endocrine System ,Immunodominance ,Mass spectrometry ,Immune Activation ,Interferon-gamma ,Islets of Langerhans ,03 medical and health sciences ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,Humans ,MESH : Islets of Langerhans ,Amino Acid Sequence ,Biology ,MESH: Mass Spectrometry ,MESH: Humans ,MESH: Molecular Sequence Data ,MESH : Mice, Inbred NOD ,MESH : Reproducibility of Results ,MESH: Islets of Langerhans ,MESH : Humans ,Histocompatibility Antigens Class I ,Immunity ,MESH : Clone Cells ,Proteins ,R1 ,MESH: Glucose-6-Phosphatase ,Clone Cells ,chemistry ,Peptides ,MESH : Sequence Analysis, Protein ,MESH : Molecular Sequence Data ,Anatomy and Physiology ,Epitopes, T-Lymphocyte ,MESH: Histocompatibility Antigens Class I ,Interferon gamma ,Multidisciplinary ,T Cells ,Animal Models ,MESH : CD8-Positive T-Lymphocytes ,[SDV.IMM.IA] Life Sciences [q-bio]/Immunology/Adaptive immunology ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,MESH : Histocompatibility Antigens Class I ,Research Article ,medicine.drug ,MESH: Interferon-gamma ,Major histocompatibility complex ,MESH: Epitopes, T-Lymphocyte ,MESH : Mass Spectrometry ,Model Organisms ,MESH : Mice ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,medicine ,Animals ,Synthetic Peptide ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,MESH : Epitopes, T-Lymphocyte ,MESH: Mice ,MESH : Interferon-gamma ,030304 developmental biology ,Diabetic Endocrinology ,MESH: Clone Cells ,Reproducibility of Results ,Molecular biology ,biology.protein ,MESH : Animals ,CD8 ,030215 immunology - Abstract
International audience; 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 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.
- Published
- 2011