File(s) not publicly available

Construction of recombinant S-layer proteins (rSbsA) and their expression in bacterial ghosts : a delivery system for the nontypeable Haemophilus influenzae antigen Omp26

journal contribution
posted on 06.12.2017, 00:00 by E Riedmann, Jennelle KydJennelle Kyd, A Smith, S Gomez Gallego, K Jalava, W Lubitz, A Cripps
This study has investigated the feasibility of a combination of recombinant surface layer (S-layer) proteins and empty bacterial cell envelopes (ghosts) to deliver candidate antigens for a vaccine against nontypeable Haemophilus influenzae (NTHi) infections. The S-layer gene sbsA from Bacillus stearothermophilus PV72 was used for the construction of fusion proteins. Fusion of maltose binding protein (MBP) to the N-terminus of SbsA allowed expression of the S-layer in the periplasm of Escherichia coli. The outer membrane protein (Omp) 26 of NTHi was inserted into the N-terminal and C-terminal regions of SbsA. The presence of the fused antigen Omp26 was demonstrated by Western blot experiments using anti-Omp26 antisera. Electron microscopy showed that the recombinant SbsA maintained the ability to self-assemble into sheet-like and cylindrical structures. Recombinant E. coli cell envelopes (ghosts) were produced by the expression of SbsA/Omp26 fusion proteins prior to gene E-mediated lysis. Intraperitoneal immunization with these recombinant bacterial ghosts induced an Omp26-specific antibody response in BALB/c mice. These results demonstrate that the NTHi antigen, Omp26, was expressed in the S-layer self-assembly product and this construct was immunogenic for Omp26 when administered to mice in bacterial cell envelopes.

History

Volume

37

Issue

2-3

Start Page

185

End Page

192

Number of Pages

8

eISSN

1574-695X

ISSN

0928-8244

Location

United Kingdom

Publisher

Wiley-Blackwell Publishing

Language

en-aus

Peer Reviewed

Yes

Open Access

No

External Author Affiliations

Bird-C GmbH & CoKG (Austria); Universität Wien; University of Canberra;

Era Eligible

Yes

Journal

FEMS immunology and medical microbiology.