title: Bioinspired Design and Oriented Synthesis of Immunogenic Site-Specifically Penicilloylated Peptides.
abstract: β-Lactam antibiotics allergy is recognized as a public health concern. By covalently binding to serum proteins, penicillins are known to form immunogenic complexes. The latter are recognized and digested by antigen-presenting cells into drug-hapten peptides leading to the immunization of treated persons and IgE-mediated hypersensitivity reactions encompassing anaphylaxis. If type I allergic reactions to drugs are often unpredictable, they are known to be dependent on CD4+T-cells. This fundamental study revisits the chemical basis of the benzylpenicillin (BP) allergy with the aim of identifying immunologically relevant biomimetic benzylpenicilloylated peptides through the analysis of BP-conjugated human serum albumin (BP-HSA) profile and the evaluation of the naı̈ve CD4+T-cell responses to candidate BP-HSA-derived peptides. The chemical structures of BP-HSA bioconjugates synthesized in vitro at both physiological and basic pH were investigated by mass spectrometry. From the ten most representative lysine residues grafted by BP-hapten, HSA-bioinspired 15-mer peptide sequences were designed and the potential T-cell epitope profile of each peptide was predicted using two complementary in silico approaches, i.e., HLA class II binding prediction tools from the Immune Epitope Database and Analysis Resource (IEDB) and computational alanine scanning mutagenesis. Twelve structurally diversified benzylpenicilloylated peptides (BP-Ps) were selected and synthesized with the aid of a flexible synthesis pathway using an original benzylpenicilloylated lysine monomer as common precursor. In order to corroborate their predicted "epitope" profile, the naı̈ve CD4+T-cell response specific to BP was evaluated through a coculture approach. To our knowledge, this study showed for the first time the ability of bioinspired peptides structurally stemming from BP-HSA to be recognized by naı̈ve CD4+T-cells thus identifying a pre-existing T-cell repertoire for penicillin molecules bound to proteins. It also established a promising model approach expandable to other most frequently used penicillin classes of antibiotics to reveal biomimetic drug-modified antigenic peptides relevant for qualitative and quantitative drug allergy studies.
authors.affiliation: BioCIS, Université Paris-Sud, CNRS, Université Paris-Saclay , Châtenay-Malabry, F-92296, France.
authors.forename: Noémie
authors.initials: N
authors.lastname: Scornet
authors.affiliation: BioCIS, Université Paris-Sud, CNRS, Université Paris-Saclay , Châtenay-Malabry, F-92296, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Delarue-Cochin
authors.affiliation: INSERM UMR 996, Université Paris-Sud, Université Paris-Saclay , Châtenay-Malabry, F-92296, France.
authors.forename: Marie Eliane
authors.initials: ME
authors.lastname: Azoury
authors.affiliation: Stallergenes Greer , 6 rue Alexis de Tocqueville, Antony, 92160, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Le Mignon
authors.affiliation: PRABI-LG, UMR CNRS 5305 (LBTI) , Lyon, 69367, France.|UFR Pharmacie , Lyon, 69367, France.
authors.forename: Julie-Anne
authors.initials: JA
authors.lastname: Chemelle
authors.affiliation: Stallergenes Greer , 6 rue Alexis de Tocqueville, Antony, 92160, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Nony
authors.affiliation: CEA, Institut de Biologie et de Technologies, Université Paris-Saclay , Gif-sur-Yvette, 91190, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Maillère
authors.affiliation: PRABI-LG, UMR CNRS 5305 (LBTI) , Lyon, 69367, France.|UFR Pharmacie , Lyon, 69367, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Terreux
authors.affiliation: INSERM UMR 996, Université Paris-Sud, Université Paris-Saclay , Châtenay-Malabry, F-92296, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Pallardy
authors.affiliation: BioCIS, Université Paris-Sud, CNRS, Université Paris-Saclay , Châtenay-Malabry, F-92296, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Joseph
mesh_terms: D000595:Amino Acid Sequence; D032701:Biomimetics; D060326:Chemistry Techniques, Synthetic; D003198:Computer Simulation; D015195:Drug Design; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D006241:Haptens / Q000737:chemistry; D006801:Humans; D007114:Immunization; D007073:Immunoglobulin E / Q000276:immunology; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D010400:Penicillin G / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000276:immunology*; D011487:Protein Conformation; D012709:Serum Albumin / Q000737:chemistry
chemical_list: D000939:Epitopes; D006241:Haptens; D010455:Peptides; D012709:Serum Albumin; D007073:Immunoglobulin E; D008239:Lysine; D010400:Penicillin G