title: Treatment of both native and deamidated gluten peptides with an endo-peptidase from Aspergillus niger prevents stimulation of gut-derived gluten-reactive T cells from either children or adults with celiac disease.
abstract: Celiac disease (CD) is characterized by an inappropriate immunological reaction against gluten driven by gluten-specific CD4+ T cells. We screened 25 proteases and tested 10 for their potential to degrade gluten in vitro. Five proteases were further tested for their ability to prevent the proliferative response by a gluten-specific CD4+ T cell clone and seven gluten-reactive T cell lines to protease-digested gluten peptides. A proline-specific endo-peptidase from Aspergillus niger (AnP2) was particularly efficient at diminishing proliferation after stimulation with cleaved antigen, and could completely block the response against both native and deamidated gluten peptides. We found that AnP2 was efficient down to a 1:64 protease:substrate ratio (w:w). When AnP2 was tested in assays using seven gluten-reactive T cell lines from individual CD patients (three adults and four children), the response to gluten was diminished in all cases. Our study indicates a therapeutic benefit of AnP2 to CD patients.
authors.affiliation: Hans Christian Andersen Children's Hospital, Odense University Hospital, University of Southern Denmark, Denmark; Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, Denmark. Electronic address: htoft-hansen@health.sdu.dk.
authors.forename: Henrik
authors.initials: H
authors.lastname: Toft-Hansen
authors.affiliation: Hans Christian Andersen Children's Hospital, Odense University Hospital, University of Southern Denmark, Denmark; Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, Denmark.
authors.forename: Karina S
authors.initials: KS
authors.lastname: Rasmussen
authors.affiliation: Hans Christian Andersen Children's Hospital, Odense University Hospital, University of Southern Denmark, Denmark; Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, Denmark.
authors.forename: Anne
authors.initials: A
authors.lastname: Staal
authors.affiliation: Immunotoxicology, Novozymes, Bagsværd, Denmark.
authors.forename: Erwin L
authors.initials: EL
authors.lastname: Roggen
authors.affiliation: Centre for Immune Regulation, University of Oslo, Norway.
authors.forename: Ludvig M
authors.initials: LM
authors.lastname: Sollid
authors.affiliation: Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, Denmark.
authors.forename: Søren T
authors.initials: ST
authors.lastname: Lillevang
authors.affiliation: Department of Clinical Immunology, Odense University Hospital, University of Southern Denmark, Denmark.
authors.forename: Torben
authors.initials: T
authors.lastname: Barington
authors.affiliation: Hans Christian Andersen Children's Hospital, Odense University Hospital, University of Southern Denmark, Denmark.
authors.forename: Steffen
authors.initials: S
authors.lastname: Husby
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002446:Celiac Disease / Q000276:immunology / Q000473:pathology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002999:Clone Cells / Q000187:drug effects / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D005656:Fungal Proteins / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D005983:Glutens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007422:Intestines / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D005656:Fungal Proteins; D010455:Peptides; D005983:Glutens; D010450:Endopeptidases
keywords: Autoimmunity;; Celiac disease;; Gluten;; Proteases