title: Function of neisserial outer membrane phospholipase a in autolysis and assessment of its vaccine potential.
abstract: Outer membrane phospholipase A (OMPLA) is an outer membrane-localized enzyme, present in many gram-negative bacterial species. It is implicated in the virulence of several pathogens. Here, we investigated the presence, function, and vaccine potential of OMPLA in the human pathogen Neisseria meningitidis. Immunoblot analysis showed the presence of OMPLA in 28 out of 33 meningococcal strains investigated. The OMPLA-negative strains all contained a pldA gene, but these alleles contained premature stop codons. All six Neisseria gonorrhoeae strains tested, but only two out of seven commensal neisserial strains investigated, expressed OMPLA, showing that OMPLA is expressed by, but not limited to, many pathogenic neisserial strains. The function of OMPLA was investigated by assessing the phenotypes of isogenic strains, expressing no OMPLA, expressing wild-type levels of OMPLA, or overexpressing OMPLA. OMPLA exhibited phospholipase activity against endogenous phospholipids. Furthermore, OMPLA was characterized as an autolysin that acted under specific conditions, such as prolonged growth of the bacteria. The vaccine potential of the protein was investigated by immunizing mice with in vitro refolded, recombinant OMPLA. High levels of antibody titers were obtained, but the murine sera were neither bactericidal nor protective. Also, convalescent patients and vaccinee sera did not contain detectable levels of anti-OMPLA antibodies, indicating that OMPLA may not be sufficiently immunogenic to be included in a meningococcal vaccine.
authors.affiliation: Department of Molecular Microbiology and Institute for Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. m.p.bos@bio.uu.nl
authors.forename: Martine P
authors.initials: MP
authors.lastname: Bos
authors.forename: Boris
authors.initials: B
authors.lastname: Tefsen
authors.forename: Pierre
authors.initials: P
authors.lastname: Voet
authors.forename: Vincent
authors.initials: V
authors.lastname: Weynants
authors.forename: Jos P M
authors.initials: JP
authors.lastname: van Putten
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D001428:Bacterial Vaccines / Q000276:immunology*; D001433:Bacteriolysis / Q000502:physiology*; D001483:Base Sequence; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000201:enzymology* / Q000276:immunology; D009345:Neisseria meningitidis / Q000201:enzymology* / Q000276:immunology; D010741:Phospholipases A / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D010741:Phospholipases A