title: Immunogenicity and efficacy of highly purified invasin complex vaccine from Shigella flexneri 2a.
abstract: Development of a subunit vaccine for shigellosis requires identification of protective antigens and delivering these antigens in a manner that stimulates immunity comparable to that induced by natural infection. The Shigella invasin complex (Invaplex) vaccine is an ion-exchange-purified extract from virulent Shigella that consists of LPS and several other proteins, including the invasins IpaB and IpaC. Intranasal delivery of Invaplex stimulates protective immunity in small animal models for shigellosis. To identify the active component(s) of Invaplex responsible for its immunogenicity and efficacy, size-exclusion chromatography (SEC) was used to separate Invaplex into several different fractions. A high-molecular mass complex with a molecular mass between 669 MDa and 2 MDa consisted primarily of LPS, IpaB and IpaC and was considered to be a highly purified (HP) form of Invaplex. Using the mouse lung model to evaluate the immunogenicity and efficacy of the SEC fractions it was clearly demonstrated that the high-molecular mass complex of the invasins and LPS was responsible for the protective capacity of parent native Invaplex. Other smaller mass SEC fractions were mostly non-immunogenic and did not stimulate solid protection. In guinea pigs, the HP Invaplex stimulated an enhanced immune response as compared to the parent Invaplex and was fully protective. Isolation and characterization of the immunogenic and protective moiety within Invaplex will allow better standardization of the Invaplex product and may allow future development of an Invaplex assembled from purified components.
authors.affiliation: Division of Bacterial and Rickettsial Diseases, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910-7500, USA.
authors.forename: K Ross
authors.initials: KR
authors.lastname: Turbyfill
authors.forename: Robert W
authors.initials: RW
authors.lastname: Kaminski
authors.forename: Edwin V
authors.initials: EV
authors.lastname: Oaks
mesh_terms: D018829:Adhesins, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D007118:Immunoassay; D007070:Immunoglobulin A / Q000096:biosynthesis / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D007637:Keratoconjunctivitis / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D008070:Lipopolysaccharides / Q000276:immunology; D008970:Molecular Weight; D022281:Shigella Vaccines / Q000276:immunology*; D012763:Shigella flexneri / Q000276:immunology*
chemical_list: D018829:Adhesins, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D008070:Lipopolysaccharides; D022281:Shigella Vaccines