title: Generation and characterization of murine antiflagellum monoclonal antibodies that are protective against lethal challenge with Pseudomonas aeruginosa.
abstract: Two murine monoclonal antibodies, IIG5 (IgG3) and IVE8 (IgG2a), that bind to Pseudomonas aeruginosa type a flagella and type b flagella, respectively, were prepared by conventional hybridoma methodology. Specificity of each monoclonal antibody for type a or type b flagella was demonstrated by enzyme-linked immunosorbent assay, indirect immunofluorescence, and immunoblotting. The percentage of P. aeruginosa isolates recognized by each monoclonal antibody was analyzed by enzyme-linked immunosorbent assay. Among a panel of 257 flagellated P. aeruginosa clinical isolates, IIG5 bound to 67.7% of the isolates and IVE8 bound to another 30.7%, for a combined coverage of 98.4%. Inhibition of motility of P. aeruginosa by the monoclonal antibodies was observed in vitro in a soft agar assay and was dose dependent. The protective efficacy of IIG5 and IVE8 was examined in a mouse burn wound sepsis model. The antiflagellum monoclonal antibodies provided specific and significant prophylactic and therapeutic protection against lethal challenge with P. aeruginosa strains.
authors.affiliation: Oncogen, Seattle, Washington 98121.
authors.forename: M J
authors.initials: MJ
authors.lastname: Rosok
authors.forename: M R
authors.initials: MR
authors.lastname: Stebbins
authors.forename: K
authors.initials: K
authors.lastname: Connelly
authors.forename: M E
authors.initials: ME
authors.lastname: Lostrom
authors.forename: A W
authors.initials: AW
authors.lastname: Siadak
mesh_terms: D000371:Agglutination; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D005260:Female; D005408:Flagellin / Q000032:analysis / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009068:Movement; D011552:Pseudomonas Infections / Q000517:prevention & control; D011550:Pseudomonas aeruginosa / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D005408:Flagellin