title: Effective mucosal immunization against respiratory syncytial virus using purified F protein and a genetically detoxified cholera holotoxin, CT-E29H.
abstract: We exploited the powerful adjuvant properties of cholera holotoxin (CT) to create a mucosally administered subunit vaccine against respiratory syncytial virus (RSV). A genetically detoxified mutant CT with an E to H substitution at amino acid 29 of the CT-A1 subunit (CT-E29H) was compared to wild type CT for toxicity and potential use as an intranasal (IN) adjuvant for the natural fusion (F) protein of RSV. When compared to CT the results demonstrated that: (1) CT-E29H binding to GM1 ganglioside was equivalent, (2) ADP-ribosylation of agmatine was 11.7%, and (3) toxicity was attenuated in both Y-1 adrenal (1.2%) and patent mouse gut weight assays. IN vaccination with F protein formulated with CT-E29H induced serum anti-CT and anti-F protein antibodies that were comparable to those obtained after vaccination with equivalent doses of CT. Vaccinations containing CT-E29H at doses of 0.1 microg were statistically equivalent to 1.0 microg in enhancing responses to F protein. Antigen-specific mucosal IgA and anti-RSV neutralizing antibodies were detected in nasal washes and sera, respectively, of mice that had received F protein and 0.1 or 1.0 microg of CT-E29H. Anti-F protein IgA was not detected in the nasal washes from mice IN vaccinated with 0.01 microg CT-E29H or IM with F protein adsorbed to AlOH adjuvant. In addition, the formulation of purified F protein and CT-E29H (0.1 and 1.0 microg) facilitated protection of both mouse lung and nose from live RSV challenge. Collectively, the data have important implications for vaccine strategies that use genetically detoxified mutant cholera holotoxins for the mucosal delivery of highly purified RSV antigens.
authors.affiliation: Department of Immunology Research, Wyeth-Lederle Vaccines, 211 Bailey Road, West Henrietta, NY 14586-9728, USA.
authors.forename: P W
authors.initials: PW
authors.lastname: Tebbey
authors.forename: C A
authors.initials: CA
authors.lastname: Scheuer
authors.forename: J A
authors.initials: JA
authors.lastname: Peek
authors.forename: D
authors.initials: D
authors.lastname: Zhu
authors.forename: N A
authors.initials: NA
authors.lastname: LaPierre
authors.forename: B A
authors.initials: BA
authors.lastname: Green
authors.forename: E D
authors.initials: ED
authors.lastname: Phillips
authors.forename: A R
authors.initials: AR
authors.lastname: Ibraghimov
authors.forename: J H
authors.initials: JH
authors.lastname: Eldridge
authors.forename: G E
authors.initials: GE
authors.lastname: Hancock
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D018893:Bronchoalveolar Lavage; D002772:Cholera Toxin / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015707:HN Protein; D018928:Immunity, Mucosal; D008168:Lung / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D009297:Nasal Mucosa / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000276:immunology*; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D015707:HN Protein; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D014765:Viral Vaccines; C116204:attachment protein G; D002772:Cholera Toxin