Bactericidal activity of antibodies elicited against the Neisseria meningitidis 37-kDa ferric binding protein (FbpA) with different adjuvants

Among 25 serogroup BNeisseria meningitidisclinical isolates, we identified four (16%) with high factor H binding protein (FHbp) expression that were resistant to complement-mediated bactericidal activity of sera from mice immunized with recombinant FHbp vaccines. Two of the four isolates had evidence of human FH-dependent complement downregulation independent of FHbp. Since alternative complement pathway recruitment is critical for anti-FHbp bactericidal activity, we hypothesized that in these two isolates binding of FH to ligands other than FHbp contributes to anti-FHbp bactericidal resistance. Knocking out NspA, a known meningococcal FH ligand, converted both resistant isolates to anti-FHbp susceptible isolates. The addition of a nonbactericidal anti-NspA monoclonal antibody to the bactericidal reaction also increased anti-FHbp bactericidal activity. To identify a role for FH ligands other than NspA or FHbp in resistance, we created double NspA/FHbp knockout mutants. Mutants from both resistant isolates bound 10-fold more recombinant human FH domains 6 and 7 fused to Fc than double knockout mutants prepared from two sensitive meningococcal isolates. In light of recent studies showing functional FH-PorB2 interactions, we hypothesized that PorB3 from the resistant isolates recruited FH. Allelic exchange ofporB3from a resistant isolate to a sensitive isolate increased resistance of the sensitive isolate to anti-FHbp bactericidal activity (and vice versa). Thus, some PorB3 variants functionally bind human FH, which in the presence of NspA enhances anti-FHbp resistance. Combining anti-NspA antibodies with anti-FHbp antibodies can overcome resistance. Meningococcal vaccines that target both NspA and FHbp are likely to confer greater protection than either antigen alone.

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Faculty Opinions recommendation of Neisseria meningitidis escape from the bactericidal activity of a monoclonal antibody is mediated by phase variation of lgtG and enhanced by a mutator phenotype.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1128820.585902 ◽

2008 ◽

Author(s):

William Shafer ◽

Maira Goytia

Keyword(s):

Monoclonal Antibody ◽

Neisseria Meningitidis ◽

Bactericidal Activity ◽

Phase Variation ◽

Mutator Phenotype

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Anion-independent Iron Coordination by the Campylobacter jejuni Ferric Binding Protein

Journal of Biological Chemistry ◽

10.1074/jbc.m412479200 ◽

2005 ◽

Vol 280 (10) ◽

pp. 9283-9290 ◽

Cited By ~ 31

Author(s):

Stacey A.L. Tom-Yew ◽

Diana T. Cui ◽

Elena G. Bekker ◽

Michael E.P. Murphy

Keyword(s):

Campylobacter Jejuni ◽

Binding Protein ◽

Ferric Binding Protein ◽

Iron Coordination

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Natural Serum Bactericidal Activity Against Neisseria meningitidis Isolates from Disseminated Infections in Normal and Complement-Deficient Hosts

The Journal of Infectious Diseases ◽

10.1093/infdis/152.6.1332 ◽

1985 ◽

Vol 152 (6) ◽

pp. 1332-1335 ◽

Cited By ~ 9

Author(s):

S. C. Ross ◽

H. M. Berberich ◽

P. Densen

Keyword(s):

Neisseria Meningitidis ◽

Bactericidal Activity ◽

Serum Bactericidal Activity

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Mosaic-Like Structure of Penicillin-Binding Protein 2 Gene (penA) in Clinical Isolates of Neisseria gonorrhoeae with Reduced Susceptibility to Cefixime

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.12.3744-3749.2002 ◽

2002 ◽

Vol 46 (12) ◽

pp. 3744-3749 ◽

Cited By ~ 132

Author(s):

Satoshi Ameyama ◽

Shoichi Onodera ◽

Masahiro Takahata ◽

Shinzaburo Minami ◽

Nobuko Maki ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Neisseria Gonorrhoeae ◽

Neisseria Meningitidis ◽

Binding Protein ◽

Clinical Isolates ◽

Penicillin Binding Protein ◽

Gene Sequences ◽

Neisseria Flavescens ◽

Neisseria Perflava

ABSTRACT Neisseria gonorrhoeae strains with reduced susceptibility to cefixime (MICs, 0.25 to 0.5 μg/ml) were isolated from male urethritis patients in Tokyo, Japan, in 2000 and 2001. The resistance to cephems including cefixime and penicillin was transferred to a susceptible recipient, N. gonorrhoeae ATCC 19424, by transformation of the penicillin-binding protein 2 gene (penA) that had been amplified by PCR from a strain with reduced susceptibility to cefixime (MIC, 0.5 μg/ml). The sequences of penA in the strains with reduced susceptibilities to cefixime were different from those of other susceptible isolates and did not correspond to the reported N. gonorrhoeae penA gene sequences. Some regions in the transpeptidase-encoding domain in this penA gene were similar to those in the penA genes of Neisseria perflava (N. sicca), Neisseria cinerea, Neisseria flavescens, and Neisseria meningitidis. These results showed that a mosaic-like structure in the penA gene conferred reductions in the levels of susceptibility of N. gonorrhoeae to cephems and penicillin in a manner similar to that found for N. meningitidis and Streptococcus pneumoniae.

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SUPREX (Stability of Unpurified Proteins from Rates of H/D Exchange) Analysis of the Thermodynamics of Synergistic Anion Binding by Ferric-Binding Protein (FbpA), a Bacterial Transferrin†

Biochemistry ◽

10.1021/bi0481848 ◽

2004 ◽

Vol 43 (50) ◽

pp. 15767-15774 ◽

Cited By ~ 27

Author(s):

Petra L. Roulhac ◽

Kendall D. Powell ◽

Suraj Dhungana ◽

Katherine D. Weaver ◽

Timothy A. Mietzner ◽

...

Keyword(s):

Binding Protein ◽

Anion Binding ◽

Ferric Binding Protein ◽

Bacterial Transferrin

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Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins

Infection and Immunity ◽

10.1128/iai.01815-14 ◽

2014 ◽

Vol 83 (2) ◽

pp. 730-742 ◽

Cited By ~ 16

Author(s):

Magdalena K. Bielecka ◽

Nathalie Devos ◽

Mélanie Gilbert ◽

Miao-Chiu Hung ◽

Vincent Weynants ◽

...

Keyword(s):

Neisseria Meningitidis ◽

Bactericidal Activity ◽

Binding Proteins ◽

Leader Peptide ◽

Type I ◽

Globular Domain ◽

His Tag ◽

Macrophage Infectivity Potentiator ◽

Fk506 Binding Proteins ◽

Macrophage Infectivity

A recombinant macrophage infectivity potentiator (rMIP) protein ofNeisseria meningitidisinduces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an N- or C-terminal His tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we determined that C-term His truncated rMIP (−LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not Δmipmutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the His tag at the N terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines.

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