title: Human Immunoglobulin Heavy Gamma Chain Polymorphisms: Molecular Confirmation Of Proteomic Assessment.
abstract: Immunoglobulin G (IgG) proteins are known for the huge diversity of the variable domains of their heavy and light chains, aimed at protecting each individual against foreign antigens. The IgG also harbor specific polymorphism concentrated in the CH2 and CH3-CHS constant regions located on the Fc fragment of their heavy chains. But this individual particularity relies only on a few amino acids among which some could make accurate sequence determination a challenge for mass spectrometry-based techniques.The purpose of the study was to bring a molecular validation of proteomic results by the sequencing of encoding DNA fragments. It was performed using ten individual samples (DNA and sera) selected on the basis of their Gm (gamma marker) allotype polymorphism in order to cover the main immunoglobulin heavy gamma (IGHG) gene diversity. Gm allotypes, reflecting part of this diversity, were determined by a serological method. On its side, the IGH locus comprises four functional IGHG genes totalizing 34 alleles and encoding the four IgG subclasses. The genomic study focused on the nucleotide polymorphism of the CH2 and CH3-CHS exons and of the intron. Despite strong sequence identity, four pairs of specific gene amplification primers could be designed. Additional primers were identified to perform the subsequent sequencing. The nucleotide sequences obtained were first assigned to a specific IGHG gene, and then IGHG alleles were deduced using a home-made decision tree reading of the nucleotide sequences. IGHG amino acid (AA) alleles were determined by mass spectrometry. Identical results were found at 95% between alleles identified by proteomics and those deduced from genomics. These results validate the proteomic approach which could be used for diagnostic purposes, namely for a mother-and-child differential IGHG detection in a context of suspicion of congenital infection.
authors.affiliation: From the ‡Institut de Recherche pour le Développement, UMR 216 Mère et enfant face aux infections tropicales, Paris, France.|§COMUE Sorbonne Paris Cité, Faculté de Pharmacie, Université Paris Descartes, Paris, France.|¶¶Magalie Dambrun, Célia Dechavanne and Alexandra Emmanuel contributed equally to this work.
authors.forename: Magalie
authors.initials: M
authors.lastname: Dambrun
authors.affiliation: From the ‡Institut de Recherche pour le Développement, UMR 216 Mère et enfant face aux infections tropicales, Paris, France.|§COMUE Sorbonne Paris Cité, Faculté de Pharmacie, Université Paris Descartes, Paris, France.|¶¶Magalie Dambrun, Célia Dechavanne and Alexandra Emmanuel contributed equally to this work.
authors.forename: Célia
authors.initials: C
authors.lastname: Dechavanne
authors.affiliation: From the ‡Institut de Recherche pour le Développement, UMR 216 Mère et enfant face aux infections tropicales, Paris, France.|§COMUE Sorbonne Paris Cité, Faculté de Pharmacie, Université Paris Descartes, Paris, France.|¶ESPCI Paris, PSL Research University, Spectrométrie de Masse Biologique et Protéomique (SMBP), CNRS USR 3149, Paris, France.|¶¶Magalie Dambrun, Célia Dechavanne and Alexandra Emmanuel contributed equally to this work.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Emmanuel
authors.affiliation: From the ‡Institut de Recherche pour le Développement, UMR 216 Mère et enfant face aux infections tropicales, Paris, France.|§COMUE Sorbonne Paris Cité, Faculté de Pharmacie, Université Paris Descartes, Paris, France.
authors.forename: Florentin
authors.initials: F
authors.lastname: Aussenac
authors.affiliation: ‖Plate-forme protéomique de l'Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Marjorie
authors.initials: M
authors.lastname: Leduc
authors.affiliation: ¶ESPCI Paris, PSL Research University, Spectrométrie de Masse Biologique et Protéomique (SMBP), CNRS USR 3149, Paris, France.
authors.forename: Chiara
authors.initials: C
authors.lastname: Giangrande
authors.affiliation: ¶ESPCI Paris, PSL Research University, Spectrométrie de Masse Biologique et Protéomique (SMBP), CNRS USR 3149, Paris, France.
authors.forename: Joëlle
authors.initials: J
authors.lastname: Vinh
authors.affiliation: **Laboratoire d'Anthropologie Moléculaire et Imagerie de Synthèse, UMR 5288, CNRS et Université Paul Sabatier Toulouse III, Toulouse, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Dugoujon
authors.affiliation: ‡‡IMGT®, the international ImMunoGeneTics information system®, Laboratoire d'ImmunoGénétique Moléculaire, LIGM, Institut de Génétique Humaine, IGH, UMR 9002, CNRS et Université de Montpellier, Montpellier, France.|§§Institut Universitaire de France, Paris, France.
authors.forename: Marie-Paule
authors.initials: MP
authors.lastname: Lefranc
authors.affiliation: ‖Plate-forme protéomique de l'Université Paris Descartes, Sorbonne Paris Cité, Paris, France.|‖‖François Guillonneau and Florence Migot-Nabias contributed equally to this work.
authors.forename: François
authors.initials: F
authors.lastname: Guillonneau
authors.affiliation: From the ‡Institut de Recherche pour le Développement, UMR 216 Mère et enfant face aux infections tropicales, Paris, France; florence.migot-nabias@ird.fr.|§COMUE Sorbonne Paris Cité, Faculté de Pharmacie, Université Paris Descartes, Paris, France.|‖‖François Guillonneau and Florence Migot-Nabias contributed equally to this work.
authors.forename: Florence
authors.identifier: 0000-0001-9982-594X
authors.initials: F
authors.lastname: Migot-Nabias
mesh_terms: D000483:Alleles; D002648:Child; D002675:Child, Preschool; D030562:Databases, Protein; D005260:Female; D006801:Humans; D007129:Immunoglobulin Gm Allotypes; D007142:Immunoglobulin gamma-Chains / Q000235:genetics*; D008297:Male; D013058:Mass Spectrometry; D010455:Peptides / Q000378:metabolism; D011110:Polymorphism, Genetic; D040901:Proteomics / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D007129:Immunoglobulin Gm Allotypes; D007142:Immunoglobulin gamma-Chains; D010455:Peptides