title: Utilization of soluble fusion proteins for induction of T cell proliferation.
abstract: A peptide display library was evaluated as a means to identify peptide binding motifs for class II molecules. Peptides expressed as part of a soluble fusion protein with a maltose binding protein (malE) were produced by Escherichia coli. Constructs containing the high-affinity binding influenza hemagglutinin peptide 307W-319 (mal-HA) or the low-affinity binding tetanus toxoid peptide 830-843 (mal-TT) were used as controls. mal-HA, but not mal-TT, inhibited synthetic biotinylated-HA peptide from binding to purified DR4 Dw4 molecules in a dose-dependent manner. The fusion-peptide presentation system was also evaluated for its ability to induce antigen-specific T cell proliferation. DR4 Dw4+ B cells pulsed with mal-HA, but not mal-TT, induced dose-dependent proliferation of an HA-specific DR4 Dw4-restricted T cell line to the same extent as synthetic HA peptide. Using this type of peptide display library, it may be possible to determine the antigenic specificity of T cell clones isolated from patients with autoimmune diseases.
authors.affiliation: Department of Immunology and Glycobiology, Monsanto Corporate Research/G. D. Searle, St. Louis, Missouri 63198, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Kirschmann
authors.forename: P A
authors.initials: PA
authors.lastname: De Ciechi
authors.forename: C P
authors.initials: CP
authors.lastname: Bono
authors.forename: M L
authors.initials: ML
authors.lastname: Zacheis
authors.forename: B D
authors.initials: BD
authors.lastname: Schwartz
authors.forename: S L
authors.initials: SL
authors.lastname: Woulfe
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D017951:Antigen Presentation; D001402:B-Lymphocytes / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D006681:HLA-D Antigens / Q000276:immunology*; D006684:HLA-DR Antigens / Q000276:immunology; D059810:HLA-DR Serological Subtypes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology*; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D013745:Tetanus Toxoid / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D006681:HLA-D Antigens; D006684:HLA-DR Antigens; D059810:HLA-DR Serological Subtypes; C080766:HLA-DR11 antigen; C053495:HLA-Dw4 antigen; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010446:Peptide Fragments; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; D013745:Tetanus Toxoid; C081942:influenza hemagglutinin (307-319); C461599:maltose transport system, E coli; C093151:tetanus toxoid (830-843)