title: Peanut T-cell epitope discovery: Ara h 1.
abstract: Identification of potential T-cell epitopes in the peanut major allergens is essential for development of peptide-based immunotherapy. Traditional methods of T-cell epitope discovery use overlapping short peptides spanning the full length of the protein in T-cell proliferation assays. Because large proteins, such as Ara h 1, require a large number of peptides, this limits screening to a small number of allergic subject-derived T-cell lines.
  
  
  
  We sought to identify candidate peptides of Ara h 1 that display promiscuous binding to MHC class II and induce TH2 cytokine production by T cells.
  
  
  
  In silico MHC class II binding prediction was performed with NetMHCIIpan 2.0 (peptide length, 15; 1-mer offset) and the most abundant class II alleles in the North American population and with an in vitro MHC class II peptide reporter assay performed in parallel, which used synthetic 15-mer peptides offset by 5 mer spanning the protein. High-resolution MHC class II typing and a T-cell proliferation assay using preselected peptides were performed with PBMCs from 98 subjects with peanut allergy and 14 healthy control subjects. IL-4, IL-13, IL-5, IFN-γ, and TNF-α levels were measured in culture supernatants.
  
  
  
  Thirty-six Ara h 1 peptides were identified by using in silico predictions and MHC class II binding assays. In combination with T-cell proliferation and cytokines secreted in T-cell assays, we have identified 4 vaccine candidate Ara h 1 peptides.
  
  
  
  Preselection of peptides by using in silico and in vitro approaches in combination with conventional methods appears to be an effective strategy for identifying peanut T-cell peptide vaccine candidates.
authors.affiliation: Division of Allergy and Immunology, Department of Medicine, Montefiore Medical Center, Bronx, NY.
authors.forename: Manish
authors.initials: M
authors.lastname: Ramesh
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY; Division of Pediatric Allergy and Immunology, Department of Pediatrics, Faculty of Medicine, Prince of Songkla University, Hat-Yai, Songkhla, Thailand.
authors.forename: Araya
authors.initials: A
authors.lastname: Yuenyongviwat
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY; Department of Allergy and Clinical Immunology, 424 General Military Training Hospital, Thessaloniki, Greece.
authors.forename: George N
authors.initials: GN
authors.lastname: Konstantinou
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY; University of Tennessee Health Science Center College of Medicine, Memphis, Tenn.
authors.forename: Jay
authors.initials: J
authors.lastname: Lieberman
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY; Immunology Department, CDB Hospital Clinic de Barcelona, Universitat de Barcelona, Barcelona, Spain.
authors.forename: Mariona
authors.initials: M
authors.lastname: Pascal
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY.
authors.forename: Madhan
authors.initials: M
authors.lastname: Masilamani
authors.affiliation: Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY. Electronic address: hugh.sampson@mssm.edu.
authors.forename: Hugh A
authors.initials: HA
authors.lastname: Sampson
mesh_terms: D000293:Adolescent; D000918:Antibody Specificity / Q000276:immunology; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology*; D010367:Arachis / Q000276:immunology*; D016022:Case-Control Studies; D002648:Child; D016207:Cytokines / Q000378:metabolism; D018984:Epitopes, T-Lymphocyte / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D008297:Male; D008565:Membrane Proteins; D021183:Peanut Hypersensitivity / Q000175:diagnosis / Q000276:immunology / Q000628:therapy; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology*; D011485:Protein Binding
chemical_list: D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D006023:Glycoproteins; D000949:Histocompatibility Antigens Class II; D008565:Membrane Proteins; D010455:Peptides; D010940:Plant Proteins; D007073:Immunoglobulin E
keywords: Ara h 1; food allergy; peanut; peptide epitope