title: Induction of immune responses by purified outer membrane protein complexes from Neisseria meningitidis.
abstract: A broad-spectrum vaccine against disease caused by serogroup B of Neisseria meningitidis is still a challenge due to antigenic variability. In the present study outer membrane protein complexes and their components were analysed using non-denaturing 2D electrophoresis and identified using LC/MS-MS and MALDI-TOF. Outer membrane protein complexes were purified from both the wild-type strain H44/76 and their knock-out mutants lacking PorA, PorB, RmpM or FetA. The immune responses elicited by the whole outer membrane vesicles (OMV) and the purified complexes were analysed for bactericidal activity, antibody surface binding, antibody-mediated C3b/iC3b deposition, membrane attack complex (MAC) deposition and induction of opsonophagocytosis, both on the homologous and several heterologous strains. The main antigenic complexes found were homomeric, formed by the 60 kDa chaperonin (MSP63) or PorB, or heteromeric, formed by different combinations of PorA, PorB and/or RmpM. The lack of some of these proteins in the OMVs from the knock-out mutants did not affect significantly the immune responses analysed except MAC, which was significantly reduced in the anti-PorA- and anti-PorB- sera, and bactericidal activity, which was absent in the anti-PorA- serum. The sera against purified native complexes showed variable activities against the homologous strain, with greatest responses observed for anti-chaperonin and anti-PorA/PorB/RmpM sera. When tested against heterologous strains, the only anti-complex serum showing consistent responses was that against the 60 kDa chaperonin. The comparison of the responses elicited by the different sera suggests an important role of conformational epitopes, present only in native complexes, in the induction of more effective responses against N. meningitidis.
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Farmacia, Campus Sur, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Marzoa
authors.forename: S
authors.initials: S
authors.lastname: Sánchez
authors.forename: L
authors.initials: L
authors.lastname: Costoya
authors.forename: E
authors.initials: E
authors.lastname: Diéguez-Casal
authors.forename: P
authors.initials: P
authors.lastname: Freixeiro
authors.forename: C
authors.initials: C
authors.lastname: Brookes
authors.forename: L
authors.initials: L
authors.lastname: Allen
authors.forename: S
authors.initials: S
authors.lastname: Taylor
authors.forename: A R
authors.initials: AR
authors.lastname: Gorringe
authors.forename: C M
authors.initials: CM
authors.lastname: Ferreirós
authors.forename: M T
authors.initials: MT
authors.lastname: Criado
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood* / Q000276:immunology; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015938:Complement Membrane Attack Complex; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000276:immunology; D018922:HL-60 Cells; D006801:Humans; D007106:Immune Sera; D007114:Immunization; D022401:Meningococcal Vaccines / Q000008:administration & dosage / Q000276:immunology*; D051379:Mice; D008808:Mice, Inbred CBA; D009154:Mutation; D009345:Neisseria meningitidis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010587:Phagocytosis; D011487:Protein Conformation
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D015938:Complement Membrane Attack Complex; D000939:Epitopes; D007106:Immune Sera; D022401:Meningococcal Vaccines