title: Meningococcal lipopolysaccharide (LPS)-derived oligosaccharide-protein conjugates evoke outer membrane protein- but not LPS-specific bactericidal antibodies in mice: influence of adjuvants.
abstract: Meningococcal lipopolysaccharide (LPS)-derived oligosaccharides (OS) were coupled to tetanus toxoid (TT) and purified P1.7,16 outer membrane proteins (OMP). The immunogenicities of the conjugates with and without the addition of the adjuvant Quil A or the nonionic block polymer L121 were studied in mice. Immunotype L2 and L3,7,9 OS-TT conjugates induced immunoglobulin G (IgG) responses that were strongly augmented by Quil A and L121. These adjuvants not only enhanced the amount of IgG evoked but also shifted the IgG subclass distribution from mainly IgG1 toward the complement-activating subclasses IgG2a and IgG2b. The antibodies induced were directed against the OS part of meningococcal LPS. They were not bactericidal for group B meningococci. Both the L3,7,9 OS-P1.7,16 OMP conjugate and purified P1.7,16 OMP evoked a strong IgG response against the P1.7,16 OMP but not against the L3,7,9 LPS. These anti-OMP IgG responses were comparable to the IgG OMP-specific responses induced by the H44/76 or HIII-5 outer membrane vesicles but still did not lyse group B meningococcal strains. The IgG response evoked with OS-OMP or purified OMP consisted mainly of the IgG1 subclass, whereas the H44/76 or HIII-5 outer membrane vesicles induced high amounts of bactericidal IgG2a and IgG2b antibodies next to the IgG1 antibodies. The addition of the adjuvant Quil A or L121 to OS-OMP or OMP resulted in the induction of high levels of bactericidal anti-P1.7,16-specific OMP antibodies, as reflected by the presence of substantial amounts of IgG2a and IgG2b antibodies. These results indicate that (i) mouse anti-LPS antibodies evoked by LPS-derived OS-protein conjugates are not bactericidal for group B meningococci, (ii) extensive purification of P1.7,16 OMP can lead to the loss of the intrinsic adjuvant properties of outer membrane vesicle preparations, and (iii) the addition of suitable adjuvants restores the ability of these purified P1.7,16 OMP to induce bactericidal antibodies.
authors.affiliation: Eijkman-Winkler Laboratorium of Medical Microbiology, Experimental Medical Microbiology, Academic Hospital, Utrecht University, The Netherlands.
authors.forename: A F
authors.initials: AF
authors.lastname: Verheul
authors.forename: J A
authors.initials: JA
authors.lastname: Van Gaans
authors.forename: E J
authors.initials: EJ
authors.lastname: Wiertz
authors.forename: H
authors.initials: H
authors.lastname: Snippe
authors.forename: J
authors.initials: J
authors.lastname: Verhoef
authors.forename: J T
authors.initials: JT
authors.lastname: Poolman
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000378:metabolism*; D008581:Meningitis / Q000517:prevention & control; D051379:Mice; D009345:Neisseria meningitidis; D009844:Oligosaccharides / Q000276:immunology*; D011108:Polymers / Q000032:analysis; D062605:Quillaja Saponins; D012503:Saponins / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology; D013997:Time Factors
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008070:Lipopolysaccharides; D009844:Oligosaccharides; D011108:Polymers; D062605:Quillaja Saponins; D012503:Saponins; D013745:Tetanus Toxoid; C046386:Quil A