title: Mapping of antigenic and immunogenic sites of Haemophilus influenzae outer membrane protein P6 using synthetic lipopeptides.
abstract: The immune response to the P6 protein of Haemophilus influenzae was characterized with 24 synthetic icosapeptides and 45 dodecapeptides conjugated to the immune stimulator N-palmitoyl-S-[2,3-(bispalmitoyloxy)-(2RS)-propyl]-(R)-cysteinyl-( S)-serine . The antigenicity of these lipopeptides was investigated by enzyme-linked immunosorbent assays with rabbit anti-P6 serum. The epitopes of P6 protein were identified and localized within residues 31-46 and 59-70 and in the C-terminal part of the P6 protein. Mice were immunized with lipoicosapeptides without using additional adjuvants or carriers and the antibody titers were measured with isolated P6 protein and lipopeptides in a dot blot assay. Lipopeptides containing the sequence pattern QILDAHAA (P6 47-54) and the mouse B cell epitope GEYV (P6 43-46) induced high titers of anti P6 antibodies. These murine antibodies were able to neutralize the intact bacterium Haemophilus influenzae.
authors.affiliation: Institut für Organische Chemie, Universität Tübingen, Germany.
authors.forename: A G
authors.initials: AG
authors.lastname: Beck-Sickinger
authors.forename: H
authors.initials: H
authors.lastname: Rotering
authors.forename: K H
authors.initials: KH
authors.lastname: Wiesmüller
authors.forename: F
authors.initials: F
authors.lastname: Dorner
authors.forename: G
authors.initials: G
authors.lastname: Jung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D018073:Haemophilus Vaccines / Q000737:chemistry / Q000276:immunology*; D006193:Haemophilus influenzae / Q000737:chemistry / Q000276:immunology*; D015151:Immunoblotting; D008074:Lipoproteins / Q000032:analysis / Q000138:chemical synthesis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis* / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D014614:Vaccines, Synthetic / Q000032:analysis* / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D018073:Haemophilus Vaccines; D008074:Lipoproteins; C069990:P6 outer membrane protein, Haemophilus; D010455:Peptides; D014614:Vaccines, Synthetic