Immunogenicity of recombinant outer membrane porin protein and protective efficacy against lethal challenge with Bordetella bronchiseptica in rabbits

Abstract

Aims The outer membrane porin protein (OMPP) of Bordetella bronchiseptica is an important adhesion factor and protective immunogen. The aim of this study was to verify the immunogenicity of recombinant OMPP and its protective efficacy against a lethal challenge with B. bronchiseptica in rabbits. Methods and Results Soluble rOMPP was successfully expressed in Escherichia coli, and the purified recombinant protein was mixed with the ISA 201 VG adjuvant to prepare a subunit vaccine for B. bronchiseptica. Rabbits were immunized with the rOMPP subunit vaccine and then infected with the virulent B. bronchiseptica strain QDBb01. Rabbits immunized with the subunit vaccine were completely protected compared to the control group, and the protective effect was obviously better than that of the inactivated whole‐cell vaccine. Moreover, analysis of the immunization duration showed that the rOMPP subunit vaccine provided immune protection for at least 4 months after the second immunization. Conclusions The rOMPP subunit vaccine completely protected rabbits from a subsequent B. bronchiseptica challenge. Significance and Impact of the Study The results will provide key information for the development of a safe and effective recombinant subunit vaccine against B. bronchiseptica in rabbits.

A Marker-Free Bordetella bronchiseptica aroA/bscN Double Deleted Mutant Confers Protection against Lethal Challenge

“…While several types of anti-B. bronchiseptica vaccines such as the inactivated vaccines and/or the subunit vaccines including the recombinant OMPs or the novel adjuvant BfcA are commercially available or reported in laboratory phase [10,15,16], attenuated live vaccines are still proposed to be the preferable vaccines [17]. First, compared to the inactivated B. bronchiseptica vaccines and/or subunit vaccines which are often administered intramuscularly, the attenuated live B. bronchiseptica vaccines are administered through intranasal inoculation, which represents an easier way for administration [6,[17][18][19].…”

Immunogenic characteristics of the outer membrane phosphoporin as a vaccine candidate against Klebsiella pneumoniae

“…The molecular weight and purity of the PhoE was examined by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), after which the protein bands were transferred onto NC membranes (Sangon Biotech, Shanghai, China). Western blot was conducted as described in another study [ 24 ]. For blotting, the purified protein was detected using a primary monoclonal antibody against the His tag (Roche, Basel, Switzerland) with a 1:5000 dilution and a second antibody horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Sangon Biotech, Shanghai, China), diluted 1:10 000.…”

“…Before conducting the immunization experiment, the mouse sera were confirmed to be negative for antibodies against KP by indirect ELISA (iELISA, described in following section). Mice in groups 1 and 2 were subcutaneously injected with 100 µL of a solution containing 10 μg of purified PhoE mixed with Freund's adjuvant at a 1:1 (v/v) ratio or inactivated whole bacteria (CVCC4080, 1 × 10 8 CFU/mL, IWB) mixed with Freund's adjuvant according to previous studies [ 24 , 25 ]. As a control, mice in group 3 were mock-immunized with PBS only.…”

Reduction of endotoxicity in Bordetella bronchiseptica by lipid A engineering: Characterization of lpxL1 and pagP mutants

“…In addition, the presence of several proteins that are considered as potential vaccine antigens, i.e. the iron-regulated siderophore receptor FauA [37,38] the major porin OmpP [39], and the Bordetella resistance-to-killing autotransporter BrkA [40], was investigated by Western blotting. All these proteins were equally well detectable in the outer-membrane preparations of both strains (Figure 9b).…”