title: Immunization with recombinant Opc outer membrane protein from Neisseria meningitidis: influence of sequence variation and levels of expression on the bactericidal immune response against meningococci.
abstract: The opc gene from Neisseria meningitidis was cloned into the pRSETA vector, and recombinant protein was expressed at high levels in Escherichia coli. The protein was readily purified by affinity chromatography and used for immunization with conventional Al(OH)3 adjuvant or after incorporation into liposomes and Zwittergent micelles. The resulting sera were analyzed for their ability to recognize purified recombinant protein and "native" protein in an enzyme immunoassay with outer membranes and by whole-cell immunofluorescence. Immunization with Al(OH)3 induced high levels of antibodies which reacted with the purified protein but did not recognize whole cells. In contrast, liposomes and micelles induced antibodies which reacted with the native protein in whole cells. The addition of monophosphoryl lipid A (MPLA) to either liposomes or micelle preparations increased the magnitude of the immune response and induced a wider range of immunoglobulin subclasses. This was associated with the ability of the sera to induce complement-mediated killing of the homologous strain. The most effective bactericidal activity was observed with Opc protein incorporated into liposomes containing MPLA. The magnitude of the bactericidal effect was strongly influenced by the level of expression of the Opc protein and was abolished by limited variation in the sequence of the protein expressed by heterologous strains.
authors.affiliation: Molecular Microbiology Group, Division of Cell and Molecular Medicine, University of Southampton Medical School, Southampton General Hospital, United Kingdom.
authors.forename: K A
authors.initials: KA
authors.lastname: Jolley
authors.forename: L
authors.initials: L
authors.lastname: Appleby
authors.forename: J C
authors.initials: JC
authors.lastname: Wright
authors.forename: M
authors.initials: M
authors.lastname: Christodoulides
authors.forename: J E
authors.initials: JE
authors.lastname: Heckels
mesh_terms: D000276:Adjuvants, Immunologic; D000536:Aluminum Hydroxide / Q000276:immunology; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001770:Blood Bactericidal Activity; D003001:Cloning, Molecular; D014644:Genetic Variation; D005822:Genetic Vectors; D007106:Immune Sera; D007114:Immunization; D008081:Liposomes / Q000276:immunology; D008589:Meningococcal Infections / Q000382:microbiology / Q000517:prevention & control*; D022401:Meningococcal Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007106:Immune Sera; D008081:Liposomes; D022401:Meningococcal Vaccines; D011994:Recombinant Proteins; C074517:opc protein, bacteria; D000536:Aluminum Hydroxide