title: Complement activation and formation of the membrane attack complex on serogroup B Neisseria meningitidis in the presence or absence of serum bactericidal activity.
abstract: Encapsulated meningococci are complement sensitive only in the presence of bactericidal antibodies by yet-unexplored mechanisms. The objective of this study was to investigate the involvement of major bacterial surface constituents on complement activation and membrane attack complex (MAC) formation on serogroup B meningococci in the presence or absence of antibody-dependent serum bactericidal activity (SBA). The strains used were the encapsulated H44/76, five of its variants differing in capsulation and expression of the class 1 porin (PorA), and its lipopolysaccharide (LPS)-deficient isogenic mutant (LPS(-)) pLAK33. Two normal sera, one with high SBA (SBA(+)) and one with no bactericidal activity (SBA(-)) against H44/76 as well as an a-gamma-globulinemic serum were used for sensibilization of the bacteria. C3b and iC3b deposition on H44/76, its unencapsulated variant v24, and pLAK33 was similar in SBA(+) and SBA(-) serum, and no difference was present between the strains. MAC deposition on H44/76 was higher in SBA(+) serum than in SBA(-) serum and the a-gamma-globulinemic serum. The amounts of C3b on H44/76, v24, and pLAK33 in the a-gamma-globulinemic serum were also not different, indicating immunoglobulin G (IgG)- and LPS-independent complement activation. H44/76 PorA(+) and its PorA(-) variant and the v24 PorA(+) and its PorA(-) variant incubated in SBA(-) serum induced comparable amounts of MAC, despite their different serum sensitivities. Complement formation on the surface of the bacteria occurred almost exclusively via the classical pathway, but the considerable amounts of Bb measured in the serum indicated alternative pathway activation in the fluid phase. We conclude that complement deposition on meningococci is, for the most part, independent of classical pathway IgG and is not influenced by the presence of PorA or LPS on the meningococcal surface. Addition of an anti-PorA chimeric antibody to the nonbactericidal normal serum, while promoting a dose-related bacterial lysis, did not influence the amounts of C3b, iC3b, and MAC formed on the bacterial surface. These findings support the hypothesis that proper MAC insertion rather than the quantity of MAC formed on the bacterial surface is of importance for efficient lysis of meningococci.
authors.affiliation: Department of Medical Microbiology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. m.apiranthitou@amc.uva.nl
authors.forename: M
authors.initials: M
authors.lastname: Drogari-Apiranthitou
authors.forename: E J
authors.initials: EJ
authors.lastname: Kuijper
authors.forename: N
authors.initials: N
authors.lastname: Dekker
authors.forename: J
authors.initials: J
authors.lastname: Dankert
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology; D003167:Complement Activation / Q000276:immunology*; D003179:Complement C3b / Q000276:immunology*; D015938:Complement Membrane Attack Complex / Q000276:immunology*; D006801:Humans; D009345:Neisseria meningitidis / Q000276:immunology*; D018272:Porins / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D015938:Complement Membrane Attack Complex; D018272:Porins; C057699:porin protein, Neisseria; D003179:Complement C3b