Factor H-Binding Protein Is Important for Meningococcal Survival in Human Whole Blood and Serum and in the Presence of the Antimicrobial Peptide LL-37

Seib, Kate L.; Serruto, Davide; Oriente, Francesca; Delany, Isabel; Adu‐Bobie, Jeannette; Veggi, Daniele; Aricò, Beatrice; Rappuoli, Rino; Pizza, Mariagrazia

doi:10.1128/iai.01071-08

Cited by 100 publications

(85 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this work, the total neisserial transcriptome was analyzed in a time course experiment of incubation in an ex vivo model of bacteremia, using a tiling array. The ex vivo human whole-blood model has shown potential to examine a number of parameters that are likely to be important in the cascade of events associated with acute systemic meningococcal infection (26) and to characterize N. meningitidis factors involved in the survival of the bacterium during infection (13,52).…”

Section: Discussionmentioning

confidence: 99%

Analysis of the Regulated Transcriptome of Neisseria meningitidis in Human Blood Using a Tiling Array

et al. 2012

View full text Add to dashboard Cite

Neisseria meningitidis is the major cause of septicemia and meningococcal meningitis. During the course of infection, the bacterium must adapt to different host environments as a crucial factor for survival and dissemination; in particular, one of the crucial factors in N. meningitidis pathogenesis is the ability to grow and survive in human blood. We recently showed that N. meningitidis alters the expression of 30% of the open reading frames (ORFs) of the genome during incubation in human whole blood and suggested the presence of fine regulation at the gene expression level in order to control this step of pathogenesis. In this work, we used a customized tiling oligonucleotide microarray to define the changes in the whole transcriptional profile of N. meningitidis in a time course experiment of ex vivo bacteremia by incubating bacteria in human whole blood and then recovering RNA at different time points. The application of a newly developed bioinformatic tool to the tiling array data set allowed the identification of new transcripts-small intergenic RNAs, cis-encoded antisense RNAs, mRNAs with extended 5= and 3= untranslated regions (UTRs), and operons-differentially expressed in human blood. Here, we report a panel of expressed small RNAs, some of which can potentially regulate genes involved in bacterial metabolism, and we show, for the first time in N. meningitidis, extensive antisense transcription activity. This analysis suggests the presence of a circuit of regulatory RNA elements used by N. meningitidis to adapt to proliferate in human blood that is worthy of further investigation.

show abstract

Section: Discussionmentioning

confidence: 99%

Analysis of the Regulated Transcriptome of Neisseria meningitidis in Human Blood Using a Tiling Array

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Anti-fHbp antibodies were found in convalescent sera from subjects that contracted meningococcal disease, supporting the expression of fHbps on meningococcal bacteria and their in vivo immunogenicity (8). fHbp is also an important virulence factor expressed by N. meningitidis to evade innate immune defenses by specifically binding to complement factor H (fH) (9). It was suggested that anti-fHbp antibodies can elicit protection by two mechanisms: direct complement-mediated killing of the bacterium and blocking fH binding to the bacteria to increase the susceptibility of the bacterium to killing by the alternative complement pathway (7,10).…”

Section: Introductionmentioning

confidence: 99%

The Dual Role of Lipids of the Lipoproteins in Trumenba, a Self-Adjuvanting Vaccine Against Meningococcal Meningitis B Disease

Luo

Friese

Runnels

et al. 2016

AAPS J

View full text Add to dashboard Cite

ABSTRACT.Trumenba (bivalent rLP2086) is a vaccine licensed for the prevention of meningococcal meningitis disease caused by Neisseria meningitidis serogroup B (NmB) in individuals 10-25 years of age in the USA. The vaccine is composed of two factor H binding protein (fHbp) variants that were recombinantly expressed in Escherichia coli as native lipoproteins: rLP2086-A05 and rLP2086-B01. The vaccine was shown to induce potent bactericidal antibodies against a broad range of NmB isolates expressing fHbp that were different in sequence from the fHbp vaccine antigens. Here, we describe the characterization of the vaccine antigens including the elucidation of their structure which is characterized by two distinct motifs, the polypeptide domain and the N-terminal lipid moiety. In the vaccine formulation, the lipoproteins self-associate to form micelles driven by the hydrophobicity of the lipids and limited by the size of the folded polypeptides. The micelles help to increase the structural stability of the lipoproteins in the absence of bacterial cell walls. Analysis of the lipoproteins in Toll-like receptor (TLR) activation assays revealed their TLR2 agonist activity. This activity was lost with removal of the O-linked fatty acids, similar to removal of all lipids, demonstrating that this moiety plays an adjuvant role in immune activation. The thorough understanding of the structure and function of each moiety of the lipoproteins, as well as their relationship, lays the foundation for identifying critical parameters to guide vaccine development and manufacture.

show abstract

“…fHbp is a 28 kDa surface-exposed lipoprotein and an important component of two meningococcal vaccines currently in clinical development (6)(7)(8)(9). fHbp plays a prominent role in meningococcal pathogenesis by recruiting human factor H (hfH) to the bacterial surface, thus protecting the bacterium from complement-mediated killing (10)(11)(12). fHbp is expressed by the majority of virulent meningococcal strains; its sequence displays extensive variability, allowing classification into three main variant groups (var1, -2, and -3) or two subfamilies (A and B) sharing limited cross-protection (13).…”

mentioning

confidence: 99%

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Malito

Faleri

Surdo

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

170

View full text Add to dashboard Cite

Mapping of epitopes recognized by functional monoclonal antibodies (mAbs) is essential for understanding the nature of immune responses and designing improved vaccines, therapeutics, and diagnostics. In recent years, identification of B-cell epitopes targeted by neutralizing antibodies has facilitated the design of peptide-based vaccines against highly variable pathogens like HIV, respiratory syncytial virus, and Helicobacter pylori; however, none of these products has yet progressed into clinical stages. Linear epitopes identified by conventional mapping techniques only partially reflect the immunogenic properties of the epitope in its natural conformation, thus limiting the success of this approach. To investigate antigen-antibody interactions and assess the potential of the most common epitope mapping techniques, we generated a series of mAbs against factor H binding protein (fHbp), a key virulence factor and vaccine antigen of Neisseria meningitidis. The interaction of fHbp with the bactericidal mAb 12C1 was studied by various epitope mapping methods. Although a 12-residue epitope in the C terminus of fHbp was identified by both Peptide Scanning and Phage Display Library screening, other approaches, such as hydrogen/ deuterium exchange mass spectrometry (MS) and X-ray crystallography, showed that mAb 12C1 occupies an area of ∼1,000 Å 2 on fHbp, including >20 fHbp residues distributed on both N-and C-terminal domains. Collectively, these data show that linear epitope mapping techniques provide useful but incomplete descriptions of B-cell epitopes, indicating that increased efforts to fully characterize antigen-antibody interfaces are required to understand and design effective immunogens.meningococcus | structure | surface plasmon resonance | structural mass spectrometry | antigen-antibody complex

show abstract

Factor H-Binding Protein Is Important for Meningococcal Survival in Human Whole Blood and Serum and in the Presence of the Antimicrobial Peptide LL-37

Cited by 100 publications

References 42 publications

Analysis of the Regulated Transcriptome of Neisseria meningitidis in Human Blood Using a Tiling Array

Analysis of the Regulated Transcriptome of Neisseria meningitidis in Human Blood Using a Tiling Array

The Dual Role of Lipids of the Lipoproteins in Trumenba, a Self-Adjuvanting Vaccine Against Meningococcal Meningitis B Disease

Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen

Contact Info

Product

Resources

About