Open AccessConstruction of recombinant S‐layer proteins (rSbsA) and their expression in bacterial ghosts – a delivery system for the nontypeable Haemophilus influenzae antigen Omp26
Author(s) -
Riedmann Eva M,
Kyd Jennelle M,
Smith Adam M,
GomezGallego Sara,
Jalava Katri,
Cripps Allan W,
Lubitz Werner
Publication year - 2003
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1016/s0928-8244(03)00070-1
Subject(s) - recombinant dna , biology , s layer , periplasmic space , haemophilus influenzae , microbiology and biotechnology , bacterial outer membrane , fusion protein , antigen , escherichia coli , bacterial capsule , gene , biochemistry , immunology , virulence , antibiotics
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.