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Vaccine potential and diversity of the putative cell binding factor (CBF, NMB0345/NEIS1825) protein of Neisseria meningitidis

Vaccine potential and diversity of the putative cell binding factor (CBF, NMB0345/NEIS1825) protein of Neisseria meningitidis
Vaccine potential and diversity of the putative cell binding factor (CBF, NMB0345/NEIS1825) protein of Neisseria meningitidis
The cbf gene from Neisseria meningitidis strain MC58 encoding the putative Cell Binding Factor (CBF, NMB0345/NEIS1825) protein was cloned into the pRSETA system and a ~36-kDa recombinant (r)CBF protein expressed in Escherichia coli and purified by metal affinity chromatography. High titres of rCBF antibodies were induced in mice following immunization with rCBF-saline, rCBF-Al(OH)3, rCBF-Liposomes or rCBF-Zwittergent (Zw) 3–14 micelles, both with and without incorporated monophosphoryl lipid A (MPLA) adjuvant. Anti-rCBF sera reacted in western blots of meningococcal lysates with a single protein band of molecular mass ~29.5 kDa, indicative of mature CBF protein, but did not react with a lysate of a ?nmb0345 mutant (CBF-), demonstrating specificity of the murine immune responses. CBF protein was produced by all strains of meningococci studied thus far and the protein was present on the surface of MC58 (CBF+) bacteria, but absent on ?nmb0345 mutant (CBF-) bacteria, as judged by FACS reactivity of anti-rCBF sera. Analysis of the NEIS1825 amino acid sequences from 6644 N. meningitidis isolates with defined Alleles in the pubmlst.org/Neisseria database showed that there were 141 ST types represented and there were 136 different allelic loci encoding 49 non-redundant protein sequences. Only 6/6644 (<0.1%) of N. meningitidis isolates lacked the nmb0345 gene. Amongst serogroup B isolates worldwide, ~68% and ~20% expressed CBF encoded by Allele 1 and 18 respectively, with the proteins sharing >99% amino acid identity. Murine antisera to rCBF in Zw 3–14 micelles + MPLA induced significant serum bactericidal activity (SBA) against homologous Allele 1 and heterologous Allele 18 strains, using both baby rabbit serum complement and human serum complement (h)SBA assays, but did not kill strains expressing heterologous protein encoded by Alelle 2 or 3. Furthermore, variable bactericidal activity was induced by murine antisera against different meningococcal strains in the hSBA assay, which may correlate with variable surface exposure of CBF. Regardless, the attributes of amino acid sequence conservation and protein expression amongst different strains and the ability to induce cross-strain bactericidal antibodies indicates that rCBF could be a potential meningococcal vaccine antigen and merits further testing
1932-6203
e0160403
Humbert, Maria
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Hung, Miao-Chiu
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Phillips, Renee
0d918224-043e-4492-9a20-8e7e6f632d42
Akoto, Charlene
1cd21eed-0716-468d-afa8-1c5649a70af3
Hill, Alison
de7a9d4f-7c5c-440c-9619-5b390f6347fd
Tan, Wei-Ming
4474483b-5b0f-460a-93f5-f732aba6e1f9
Heckels, John
fcfcfafe-5ca8-4728-9c5e-cb67f9af7e31
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Humbert, Maria
82134d25-24b8-4fdd-bd1c-461683b5322e
Hung, Miao-Chiu
46dfe1db-123e-427e-8b58-f7caf50a627e
Phillips, Renee
0d918224-043e-4492-9a20-8e7e6f632d42
Akoto, Charlene
1cd21eed-0716-468d-afa8-1c5649a70af3
Hill, Alison
de7a9d4f-7c5c-440c-9619-5b390f6347fd
Tan, Wei-Ming
4474483b-5b0f-460a-93f5-f732aba6e1f9
Heckels, John
fcfcfafe-5ca8-4728-9c5e-cb67f9af7e31
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078

Humbert, Maria, Hung, Miao-Chiu, Phillips, Renee, Akoto, Charlene, Hill, Alison, Tan, Wei-Ming, Heckels, John and Christodoulides, Myron (2016) Vaccine potential and diversity of the putative cell binding factor (CBF, NMB0345/NEIS1825) protein of Neisseria meningitidis. PLoS ONE, 11 (8), e0160403. (doi:10.1371/journal.pone.0160403.). (PMID:27505005)

Record type: Article

Abstract

The cbf gene from Neisseria meningitidis strain MC58 encoding the putative Cell Binding Factor (CBF, NMB0345/NEIS1825) protein was cloned into the pRSETA system and a ~36-kDa recombinant (r)CBF protein expressed in Escherichia coli and purified by metal affinity chromatography. High titres of rCBF antibodies were induced in mice following immunization with rCBF-saline, rCBF-Al(OH)3, rCBF-Liposomes or rCBF-Zwittergent (Zw) 3–14 micelles, both with and without incorporated monophosphoryl lipid A (MPLA) adjuvant. Anti-rCBF sera reacted in western blots of meningococcal lysates with a single protein band of molecular mass ~29.5 kDa, indicative of mature CBF protein, but did not react with a lysate of a ?nmb0345 mutant (CBF-), demonstrating specificity of the murine immune responses. CBF protein was produced by all strains of meningococci studied thus far and the protein was present on the surface of MC58 (CBF+) bacteria, but absent on ?nmb0345 mutant (CBF-) bacteria, as judged by FACS reactivity of anti-rCBF sera. Analysis of the NEIS1825 amino acid sequences from 6644 N. meningitidis isolates with defined Alleles in the pubmlst.org/Neisseria database showed that there were 141 ST types represented and there were 136 different allelic loci encoding 49 non-redundant protein sequences. Only 6/6644 (<0.1%) of N. meningitidis isolates lacked the nmb0345 gene. Amongst serogroup B isolates worldwide, ~68% and ~20% expressed CBF encoded by Allele 1 and 18 respectively, with the proteins sharing >99% amino acid identity. Murine antisera to rCBF in Zw 3–14 micelles + MPLA induced significant serum bactericidal activity (SBA) against homologous Allele 1 and heterologous Allele 18 strains, using both baby rabbit serum complement and human serum complement (h)SBA assays, but did not kill strains expressing heterologous protein encoded by Alelle 2 or 3. Furthermore, variable bactericidal activity was induced by murine antisera against different meningococcal strains in the hSBA assay, which may correlate with variable surface exposure of CBF. Regardless, the attributes of amino acid sequence conservation and protein expression amongst different strains and the ability to induce cross-strain bactericidal antibodies indicates that rCBF could be a potential meningococcal vaccine antigen and merits further testing

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Accepted/In Press date: 19 July 2016
Published date: 9 August 2016
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 399262
URI: https://eprints.soton.ac.uk/id/eprint/399262
ISSN: 1932-6203
PURE UUID: 5cc34c13-4223-45d0-bd37-b072d1e73a0f
ORCID for Alison Hill: ORCID iD orcid.org/0000-0001-5397-873X

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Date deposited: 11 Aug 2016 12:40
Last modified: 19 Nov 2019 01:27

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Contributors

Author: Maria Humbert
Author: Miao-Chiu Hung
Author: Renee Phillips
Author: Charlene Akoto
Author: Alison Hill ORCID iD
Author: Wei-Ming Tan
Author: John Heckels

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