Lipo-oligosaccharide immunotyping of Neisseria meningitidis by a whole-cell ELISA with monoclonal antibodies

Scholten, R. J. P. M.; Kuipers, Betsy; Valkenburg, H. A.; Dankert, J.; Zollinger, Wendell D.; Poolman, Jan

doi:10.1099/00222615-41-4-236

Cited by 117 publications

(113 citation statements)

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“…To further characterize these inner core LPS structures, wholecell lysates were prepared from various N. meningitidis strains and immunoblotting analysis performed as described previously (33) using the subset of inner core LPS MAbs (L3B5, L2-16, LPT3-1) and immunotyping MAbs (37). A summary of MAb cross-reactivity and immunotyping of 3 collections of N. meningitidis strains described in Table 2 is shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Meningococcal LPS has been classified into 12 distinct LPS immunotypes (L1 to L12), originally defined by monoclonal antibody (MAb) reactivities (37), and further defined by subsequent structural analyses of immunotypes L1/6 (5, 43), L2 (8), L3 (30), L4/7 (23), L5 (27), and L9 (12). A key determinant of the immunotyping scheme is the location of a phosphoethanolamine (PEtn) moiety on the distal inner core heptose residue (HepII) at either position 3 or position 6 or this moiety's absence.…”

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confidence: 99%

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Development, Characterization, and Functional Activity of a Panel of Specific Monoclonal Antibodies to Inner Core Lipopolysaccharide Epitopes inNeisseria meningitidis

et al. 2004

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A panel of six murine monoclonal antibodies (MAbs) recognizing inner core lipopolysaccharide (LPS) epitopes of Neisseria meningitidis was prepared and characterized in order to determine the diversity of inner core LPS glycoforms among disease and carrier isolates. Two of these MAbs, L2-16 (immunoglobulin G2b [IgG2b]) and LPT3-1 (IgG2a), together with a third, previously described MAb, L3B5 (IgG3), showed reactivity, either individually or in combination, with all except 3 of 143 disease and carriage isolates (125 of 126 strains from blood, cerebrospinal fluid, or skin biopsy samples and 15 of 17 from nasopharyngeal cultures). MAbs L3B5, L2-16, and LPT3-1 were further characterized in an indirect immunofluorescence assay. All three MAbs bound to the bacterial cell surface, findings that correlated strongly with whole-cell enzyme-linked immunosorbent assay and immunodot blots. However, in contrast to our findings with L3B5, cell surface binding of L2-16 or LPT 3-1 did not correlate with functional activity as determined by bactericidal or infant rat passive protection assays against wild-type N. meningitidis strains. These findings are provocative with respect to the requirements for protective activity of antibodies and the development of inner core LPS vaccines against invasive meningococcal disease.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Development, Characterization, and Functional Activity of a Panel of Specific Monoclonal Antibodies to Inner Core Lipopolysaccharide Epitopes inNeisseria meningitidis

et al. 2004

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“…The LOS of strain Y2220 expresses phosphoethanolamine (PEA) substitutions simultaneously at the 3 and 6 positions of HepII, a feature that is not described in any of the standard meningococcal LOS immunotypes, L1 through L12 (36). However, the inability of sialylated Y2220 to bind factor H may not be related solely to this relatively unusual HepII PEA configuration, because the LOS of a gonococcal strain, called 398079, is structurally identical to Y2220 (LNT substitution of HepI and simultaneous 3-and 6-PEA on HepII); sialylation of this gonococcal strain results in enhanced factor H binding and serum resistance (16).…”

Section: Discussionmentioning

confidence: 99%

Factor H Binding and Function in Sialylated Pathogenic Neisseriae is Influenced by Gonococcal, but Not Meningococcal, Porin

Madico

Ngampasutadol

Gulati

et al. 2007

The Journal of Immunology

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Neisseria gonorrhoeae and Neisseria meningitidis both express the lacto-N-neotetraose (LNT) lipooligosaccharide (LOS) molecule that can be sialylated. Although gonococcal LNT LOS sialylation enhances binding of the alternative pathway complement inhibitor factor H and renders otherwise serum-sensitive bacteria resistant to complement-dependent killing, the role of LOS sialylation in meningococcal serum resistance is less clear. We show that only gonococcal, but not meningococcal, LNT LOS sialylation enhanced factor H binding. Replacing the porin (Por) B molecule of a meningococcal strain (LOS sialylated) that did not bind factor H with gonococcal Por1B augmented factor H binding. Capsule expression did not alter factor H binding to meningococci that express gonococcal Por. Conversely, replacing gonococcal Por1B with meningococcal PorB abrogated factor H binding despite LNT LOS sialylation. Gonococcal Por1B introduced in the background of an unsialylated meningococcus itself bound small amounts of factor H, suggesting a direct factor H-Por1B interaction. Factor H binding to unsialylated meningococci transfected with gonococcal Por1B was similar to the sialylated counterpart only in the presence of higher (20 μg/ml) concentrations of factor H and decreased in a dose-responsive manner by ∼80% at 1.25 μg/ml. Factor H binding to the sialylated strain remained unchanged over this factor H concentration range however, suggesting that LOS sialylation facilitated optimal factor H-Por1B interactions. The functional counterpart of factor H binding showed that sialylated meningococcal mutants that possessed gonococcal Por1B were resistant to complement-mediated killing by normal human serum. Our data highlight the different mechanisms used by these two related species to evade complement.

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“…Meningococcal strains used in this study were MC58 (Virji et al, 1991), C311F3 and derivatives (Virji et al, 1993), or other strains as described by Scholten et al (1994). Meningococcal strains were grown at 37 mC in 5% CO # on Brain Heart Infusion agar 5h-GGTTTTCAAACCAGAGCC-3h 6182-6199 (BHI ; Oxoid).…”

Section: Methodsmentioning

confidence: 99%

Genetic characterization of pilin glycosylation in Neisseria meningitidis The GenBank accession number for the sequence determined in this work is AF014804.

et al. 2000

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Pili of Neisseria meningitidis are a key virulence factor, being the major adhesin of this capsulate organism and contributing to specificity for the human host. Pili are post-translationally modified by addition of an O-linked trisaccharide, Gal(β1-4)Gal(α1-3)2,4-diacetimido-2,4,6-trideoxyhexose. In a previous study the authors identified and characterized a gene, pglA, encoding a galactosyltransferase involved in pilin glycosylation. In this study a set of random genomic sequences from N. meningitidis strain MC58 was used to search for further genes involved in pilin glycosylation. Initially, an open reading frame was identified, and designated pglD (pilin glycosylation gene D), which was homologous to genes involved in polysaccharide biosynthesis. The region adjacent to this gene was cloned and nucleotide sequence analysis revealed two further genes, pglB and pglC, which were also homologous with genes involved in polysaccharide biosynthesis. Insertional mutations were constructed in pglB, pglC and pglD in N. meningitidis C311F3, a strain with well-defined LPS and pilin-linked glycan structures, to determine whether these genes had a role in the biosynthesis of either of these molecules. Analysis of these mutants revealed that there was no alteration in the phenotype of LPS in any of the mutant strains as judged by SDS-PAGE gel migration. In contrast, increased gel migration of the pilin subunit molecules of pglB, pglC and pglD mutants by Western analysis was observed. Pilin from each of the pglB, pglC and pglD mutants did not react with a terminalgalactose-specific stain, confirming that the gel migration differences were due to the alteration or absence of the pilin-linked trisaccharide structure in these mutants. In addition, antisera specific for the C311F3 trisaccharide failed to react with pilin from the pglB, pglC, pglD and galE mutants. Analysis of nucleotide sequence homologies has suggested specific roles for pglB, pglC and pglD in the biosynthesis of the 2,4-diacetimido-2,4,6-trideoxyhexose structure.

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Lipo-oligosaccharide immunotyping of Neisseria meningitidis by a whole-cell ELISA with monoclonal antibodies

Cited by 117 publications

References 5 publications

Development, Characterization, and Functional Activity of a Panel of Specific Monoclonal Antibodies to Inner Core Lipopolysaccharide Epitopes inNeisseria meningitidis

Development, Characterization, and Functional Activity of a Panel of Specific Monoclonal Antibodies to Inner Core Lipopolysaccharide Epitopes inNeisseria meningitidis

Factor H Binding and Function in Sialylated Pathogenic Neisseriae is Influenced by Gonococcal, but Not Meningococcal, Porin

Genetic characterization of pilin glycosylation in Neisseria meningitidis The GenBank accession number for the sequence determined in this work is AF014804.

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