Immunochemical and biochemical analysis of the polyvalent Pseudomonas aeruginosa vaccine PEV

Abstract

The Pseudomonas aeruginosa polyvalent vaccine PEV and its 16 constituent monovalent extracts from International Antigenic Typing System serotypes 1 through 13 and 15 through 17 (J. J. Miler, J. F. Spilsbury, R. J. Jones, E. A. Roe, and E. J. L. Lowbury, J. Med. Microbiol. 10:19-27, 1977) were subjected to biochemical analysis and to detailed immunochemical analysis with rabbit anti-PEV immunoglobulins. The results of chemical analysis, of analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis performed in conjunction with silver staining, and of analysis by crossed immunoelectrophoresis, sodium dodecyl sulfate-polyacrylamide gel-crossed immunoelectrophoresis, and Western blotting showed clearly that lipopolysaccharide was a major constituent of each monovalent extract and that it was probably the dominant antigen present in at least 15 of the 16 monovalent extracts. A 16.2-kilodalton protein, which was pronase resistant and nonsedimentable at 105,000 X g and which appeared to be biochemically and antigenically unrelated to pili, was a common although minor antigen for all extracts. Several other proteins, some of outer membrane origin, were also detected in unformalinized extracts, but these were also minor antigenic constituents of the vaccine. Neither pilin nor flagellin appeared to be major protein constituents of tested monovalent extracts, although anti-flagella antibodies could be demonstrated in rabbit anti-PEV by Western blotting. Preliminary analysis by crossed immunoelectrophoresis of serum raised in volunteers to PEV also indicated the presence therein of antibodies to lipopolysaccharide antigens.