Pseudomonas flagella Vaccine in Burns (Trauma).
Annual summary rept. 15 Jul 83-14 Jul 84,
TENNESSEE UNIV KNOXVILLE DEPT OF MICROBIOLOGY
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Contract goals are directed toward a establishing the capacity of a Pseudomonas aeruginosa flagellar vaccine to protect burned rodents both actively and passively and b understanding the importance of motility as a virulence factor in the burns animal models available. Progress has been made in isolating flagellar protein away from contaminating lipopolysaccharide. Experiments are underway to isolate large amounts of this material to establish the most efficient purification procedures. Carefully adsorbed antisera, anti-M-2 b type as well as a type, have been prepared. Contaminating lipopolysaccharide antibody was successfully eliminated because the antisera gave complete protection against challenge corresponding to the homologous flagellar antigenic type and not the 0 type or heterologous H type. Preliminary experiments indicate passive protection can be obtained against topical challenge. Experiments concerned with virulence mechanisms using four Fla- isogenic mutants and aMot- genetically constructed mutant, support the hypothesis that a functionally active flagellum is required for maximum virulence. Virulence was compared using three different challenge sites, subcutaneous, topical and intraperitoneal. Depending on the mutant and the inoculation site, decreases in virulence from two to five logs were recorded. Data from sequential tissue assays following challenge with the mutants and in protection experiments indicate that motility is particularly important in the transition from skin colonization to the circulatory system.