Identification of target antigens of antiendothelial cell antibodies in healthy individuals: A proteomic approach

Authors

  • Amélie Servettaz,

    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
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    • Both these authors contributed equally to this work.

  • Philippe Guilpain,

    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
    2. Faculté de Médecine, Pôle de Médecine Interne, Centre de Référence pour les Vascularites Nécrosantes et la Sclérodermie Systémique, Université Paris Descartes, Hôpital Cochin, Assistance Publique–Hôpitaux de Paris, Paris, France
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    • Both these authors contributed equally to this work.

  • Luc Camoin,

    1. Laboratory of Proteomics, Institut Cochin, Université Paris Descartes, CNRS - UMR 8104, Paris, France
    2. Institut Cochin, Université Paris Descartes, CNRS - UMR 8104, Paris, France
    3. INSERM U567, Paris, France
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  • Patrick Mayeux,

    1. Institut Cochin, Université Paris Descartes, CNRS - UMR 8104, Paris, France
    2. INSERM U567, Paris, France
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  • Cédric Broussard,

    1. Laboratory of Proteomics, Institut Cochin, Université Paris Descartes, CNRS - UMR 8104, Paris, France
    2. Institut Cochin, Université Paris Descartes, CNRS - UMR 8104, Paris, France
    3. INSERM U567, Paris, France
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  • Mathieu C. Tamby,

    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
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  • Nicolas Tamas,

    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
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  • Srini V. Kaveri,

    1. Centre de Recherche des Cordeliers, Université Pierre et Marie Curie -Paris 6, UMR S872, Paris, France
    2. Université Paris Descartes, UMR S872 Paris, France
    3. INSERM U872, Paris, France
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  • Loïc Guillevin,

    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
    2. Faculté de Médecine, Pôle de Médecine Interne, Centre de Référence pour les Vascularites Nécrosantes et la Sclérodermie Systémique, Université Paris Descartes, Hôpital Cochin, Assistance Publique–Hôpitaux de Paris, Paris, France
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  • Luc Mouthon Dr.

    Corresponding author
    1. Faculté de Médecine, UPRES-EA 4058, Université Paris Descartes, Paris, France
    2. Faculté de Médecine, Pôle de Médecine Interne, Centre de Référence pour les Vascularites Nécrosantes et la Sclérodermie Systémique, Université Paris Descartes, Hôpital Cochin, Assistance Publique–Hôpitaux de Paris, Paris, France
    • Laboratoire d'Immunologie, UPRES EA 4058, Pavillon Gustave Roussy, 4e étage, Hôpital Cochin, 8 rue Méchain, 75014, Paris, France Fax: +33-0-1-44-41-25-46
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Abstract

In order to identify target antigens of anti-endothelial cell (anti-EC) antibodies (AECA) in healthy individuals, we have used a proteomic approach combining 2-DE and immunoblotting. Whole cell protein extracts obtained from human umbilical vein EC (HUVEC) cultures were used as a source of antigens. Serum IgG from 12 healthy blood donors were tested at a concentration of 200 μg/mL. Targeted spots were identified by MS. The HUVEC proteome was composed of 884 protein spots. Among these, 61 ± 25.8 (mean ± SD) spots were recognized by serum IgG from healthy individuals, with marked differences from one individual to another. Among these spots, 11 were recognized by serum IgG from all healthy individuals tested. These spots corresponded to six different proteins with several spots corresponding to different isoforms of the same protein. Target antigens were: cytoskeletal proteins (β-actin, α-tubulin, and vimentin); glycolytic enzymes (glucose-3-phosphate-deshydrogenase and α-enolase); and prolyl-4-hydroxylase β subunit, a member of the disulfide isomerase family. This study shows that the repertoire of IgG AECA is heterogeneous among healthy individuals. IgG from all of the healthy individuals tested recognized a restricted set of highly conserved ubiquitous proteins playing key roles in cell biology and maintenance of homeostasis.

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