Use of a purified outer membrane protein F (porin) preparation of Pseudomonas aeruginosa as a protective vaccine in mice.
The outer membrane protein F (porin) from the PAO1 strain of Pseudomonas aeruginosa was purified by two different methods. One procedure involved separation by column chromatography of proteins extracted from isolated outer membranes, whereas the other involved extraction from gels after slab polyacrylamide gel electrophoresis of proteins extracted from cell envelopes. Both procedures yielded protein F preparations which successfully immunized mice from subsequent challenge with the PAO1 strain. The protein F preparations contained small quantities of lipopolysaccharide (LPS). This level of LPS contamination protected immunized mice from challenge with the homologous LPS serotype strain. However, immunization of mice with protein F preparations from the PAO1 strain also afforded protection against challenge with two different LPS serotype strains. This protective ability was lost when the protein F preparation was treated with papain before use as a vaccine. These observations support the conclusion that protein F has protective ability, which is not due to LPS contamination, when given as a vaccine. After immunization with the protein F preparation, mice showed an increase in antibody titer to the purified protein F preparation by enzyme-linked immunosorbent assay. Mice were protected passively by administration of rabbit antisera raised to the protein F preparation. These results indicate that the protein F preparation elicits a specific humoral antibody response in immunized animals. Our results suggest that purified protein F has potential as an effective vaccine for P. aeruginosa.
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D001426:Bacterial Proteins / Q000627:therapeutic use; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007116:Immunization, Passive; D008070:Lipopolysaccharides / Q000627:therapeutic use; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification / Q000627:therapeutic use*; D051379:Mice; D018272:Porins; D011552:Pseudomonas Infections / Q000517:prevention & control*; D011550:Pseudomonas aeruginosa / Q000032:analysis; D014611:Vaccination
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D018272:Porins