title: Neutralizing epitopes of human parainfluenza virus type 3 are conformational and cannot be imitated by synthetic peptides.
abstract: The possibility that linear epitopes on the haemagglutinin-neuraminidase (HN) surface glycoprotein of human parainfluenza virus type 3 (PIV-3) might induce neutralizing antibodies after virus infection was investigated. Thirty-seven peptides, representing 64% of the extramembranous portion of the HN molecule of PIV-3, were synthesized. Their ability to bind to 14 neutralizing murine monoclonal antibodies (mAbs) specific for HN or 26 high-titre human serum samples were tested in a direct enzyme-linked immunosorbent assay (ELISA) and in an indirect competition ELISA. None of the synthetic peptides reacted with any of the mAbs or serum samples in the direct test and none of 11 synthetic peptides tested blocked mAbs from binding to HN in the competition ELISA. These findings suggest that synthetic peptides cannot be used to imitate the known neutralizing epitopes on the HN. Analyses of reduced and non-reduced HN in ELISA and immunoblot assays confirmed that protein folding and tertiary structure are essential for epitope formation in these neutralizing sites. However, some children's sera analysed by immunoblotting contained antibodies to an uncharacterized linear epitope(s) not recognized by our panel of mAbs, raising the possibility that a neutralizing linear epitope does exist on the HN of PIV-3.
authors.affiliation: Department of Infectious Diseases, St Jude Children's Research Hospital, Memphis, Tennessee 38101-0318.
authors.forename: K J
authors.initials: KJ
authors.lastname: Henrickson
authors.forename: D W
authors.initials: DW
authors.lastname: Kingsbury
authors.forename: K L
authors.initials: KL
authors.lastname: van Wyke Coelingh
authors.forename: C W
authors.initials: CW
authors.lastname: Naeve
authors.forename: A
authors.initials: A
authors.lastname: Portner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry*; D015153:Blotting, Western; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D015707:HN Protein / Q000737:chemistry / Q000276:immunology*; D000882:Haplorhini; D006801:Humans; D008969:Molecular Sequence Data; D010224:Parainfluenza Virus 3, Human / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D011487:Protein Conformation; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D000939:Epitopes; D015707:HN Protein; D010455:Peptides; D014614:Vaccines, Synthetic