The University of Southampton
University of Southampton Institutional Repository

Structure of the recombinant Neisseria gonorrhoeae adhesin complex protein (rNg-ACP) and generation of murine antibodies with bactericidal activity against gonococci

Structure of the recombinant Neisseria gonorrhoeae adhesin complex protein (rNg-ACP) and generation of murine antibodies with bactericidal activity against gonococci
Structure of the recombinant Neisseria gonorrhoeae adhesin complex protein (rNg-ACP) and generation of murine antibodies with bactericidal activity against gonococci
Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and no effective vaccine exists currently. In this study, the structure, biological properties, and vaccine potential of the Ng-adhesin complex protein (Ng-ACP) are presented. The crystal structure of recombinant Ng-ACP (rNg-ACP) protein was solved at 1.65 Å. Diversity and conservation of Ng-ACP were examined in different Neisseria species and gonococcal isolates (https://pubmlst.org/neisseria/ database) in silico, and protein expression among 50 gonococcal strains in the Centers for Disease Control and Prevention/Food and Drug Administration (CDCP/FDA) AR Isolate Bank was examined by Western blotting. Murine antisera were raised to allele 10 (strain P9-17)-encoded rNg-ACP protein with different adjuvants and examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and a human serum bactericidal assay. Rabbit antiserum to rNg-ACP was tested for its ability to prevent Ng-ACP from inhibiting human lysozyme activity in vitro. Ng-ACP is structurally homologous to Neisseria meningitidis ACP and MliC/PliC lysozyme inhibitors. Gonococci expressed predominantly allele 10- and allele 6-encoded Ng-ACP (81% and 15% of isolates, respectively). Murine antisera were bactericidal (titers of 64 to 512, P < 0.05) for the homologous P9-17 strain and heterologous (allele 6) FA1090 strain. Rabbit anti-rNg-ACP serum prevented Ng-ACP from inhibiting human lysozyme with ∼100% efficiency. Ng-ACP protein was expressed by all 50 gonococcal isolates examined with minor differences in the relative levels of expression. rNg-ACP is a potential vaccine candidate that induces antibodies that (i) are bactericidal and (ii) prevent the gonococcus from inhibiting the lytic activity of an innate defense molecule.
2379-5042
1-25
Almonacid Mendoza, Hannia, Liliana
5ef98c79-11f8-48c8-a9dc-fdcbec45c96b
Humbert, Maria Victoria
49d3bc00-5603-4769-9eab-455a6f47c6cd
Dijokaite, Aiste
5817f906-de62-4645-8af2-edb0b29f51dd
Cleary, David W.
dd4448b2-91f4-4eb0-ad92-656a7bd3e4f1
Soo, Yiwen
1562bb70-250d-4484-8459-b5a9e54f4154
Hung, Miao-chiu
bbf6cdc6-4a7a-403c-8d18-93e1dc9c3702
Orr, Christian M.
f64259af-4120-481a-8e12-11344d005de0
Tews, Ivo
9117fc5e-d01c-4f8d-a734-5b14d3eee8dd
Machelett, Moritz M.
c9aa7658-2a08-48d6-9ce1-8c3b6d00a58c
Almonacid Mendoza, Hannia, Liliana
5ef98c79-11f8-48c8-a9dc-fdcbec45c96b
Humbert, Maria Victoria
49d3bc00-5603-4769-9eab-455a6f47c6cd
Dijokaite, Aiste
5817f906-de62-4645-8af2-edb0b29f51dd
Cleary, David W.
dd4448b2-91f4-4eb0-ad92-656a7bd3e4f1
Soo, Yiwen
1562bb70-250d-4484-8459-b5a9e54f4154
Hung, Miao-chiu
bbf6cdc6-4a7a-403c-8d18-93e1dc9c3702
Orr, Christian M.
f64259af-4120-481a-8e12-11344d005de0
Tews, Ivo
9117fc5e-d01c-4f8d-a734-5b14d3eee8dd
Machelett, Moritz M.
c9aa7658-2a08-48d6-9ce1-8c3b6d00a58c

Almonacid Mendoza, Hannia, Liliana, Humbert, Maria Victoria, Dijokaite, Aiste, Cleary, David W., Soo, Yiwen, Hung, Miao-chiu, Orr, Christian M., Tews, Ivo and Machelett, Moritz M. (2018) Structure of the recombinant Neisseria gonorrhoeae adhesin complex protein (rNg-ACP) and generation of murine antibodies with bactericidal activity against gonococci. mSphere, 10 (3), 1-25. (doi:10.1128/mSphere.00331-18).

Record type: Article

Abstract

Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and no effective vaccine exists currently. In this study, the structure, biological properties, and vaccine potential of the Ng-adhesin complex protein (Ng-ACP) are presented. The crystal structure of recombinant Ng-ACP (rNg-ACP) protein was solved at 1.65 Å. Diversity and conservation of Ng-ACP were examined in different Neisseria species and gonococcal isolates (https://pubmlst.org/neisseria/ database) in silico, and protein expression among 50 gonococcal strains in the Centers for Disease Control and Prevention/Food and Drug Administration (CDCP/FDA) AR Isolate Bank was examined by Western blotting. Murine antisera were raised to allele 10 (strain P9-17)-encoded rNg-ACP protein with different adjuvants and examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and a human serum bactericidal assay. Rabbit antiserum to rNg-ACP was tested for its ability to prevent Ng-ACP from inhibiting human lysozyme activity in vitro. Ng-ACP is structurally homologous to Neisseria meningitidis ACP and MliC/PliC lysozyme inhibitors. Gonococci expressed predominantly allele 10- and allele 6-encoded Ng-ACP (81% and 15% of isolates, respectively). Murine antisera were bactericidal (titers of 64 to 512, P < 0.05) for the homologous P9-17 strain and heterologous (allele 6) FA1090 strain. Rabbit anti-rNg-ACP serum prevented Ng-ACP from inhibiting human lysozyme with ∼100% efficiency. Ng-ACP protein was expressed by all 50 gonococcal isolates examined with minor differences in the relative levels of expression. rNg-ACP is a potential vaccine candidate that induces antibodies that (i) are bactericidal and (ii) prevent the gonococcus from inhibiting the lytic activity of an innate defense molecule.

Text
e00331-18.full - Version of Record
Available under License Creative Commons Attribution.
Download (3MB)

More information

Accepted/In Press date: 7 September 2018
e-pub ahead of print date: 10 October 2018
Published date: 10 October 2018

Identifiers

Local EPrints ID: 425428
URI: https://eprints.soton.ac.uk/id/eprint/425428
ISSN: 2379-5042
PURE UUID: f0bc3a4c-aa99-4f4d-82f0-d02d6757ed20
ORCID for Christian M. Orr: ORCID iD orcid.org/0000-0002-6137-8969
ORCID for Ivo Tews: ORCID iD orcid.org/0000-0002-4704-1139

Catalogue record

Date deposited: 19 Oct 2018 16:30
Last modified: 14 Mar 2019 01:37

Export record

Altmetrics

Contributors

Author: Hannia, Liliana Almonacid Mendoza
Author: Maria Victoria Humbert
Author: Aiste Dijokaite
Author: David W. Cleary
Author: Yiwen Soo
Author: Miao-chiu Hung
Author: Christian M. Orr ORCID iD
Author: Ivo Tews ORCID iD
Author: Moritz M. Machelett

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×