Abstract
BackgroundThere is a strong need for a recombinant subunit vaccine against fowl cholera. We used a reverse vaccinology approach to identify putative secreted or cell surface associated P. multocida proteins that may represent potential vaccine candidate antigens.Principal FindingsA high-throughput cloning and expression protocol was used to express and purify 71 recombinant proteins for vaccine trials. Of the 71 proteins tested, only one, PlpE in denatured insoluble form, protected chickens against fowl cholera challenge. PlpE also elicited comparable levels of protection in mice. PlpE was localized by immunofluorescence to the bacterial cell surface, consistent with its ability to elicit a protective immune response. To explore the role of PlpE during infection and immunity, a plpE mutant was generated. The plpE mutant strain retained full virulence for mice.ConclusionThese studies show that PlpE is a surface exposed protein and was the only protein of 71 tested that was able to elicit a protective immune response. However, PlpE is not an essential virulence factor. This is the first report of a denatured recombinant protein stimulating protection against fowl cholera.
Highlights
Bacterial vaccines can be divided into three main categories: killed whole cell vaccines, live attenuated vaccines and native or recombinant subunit vaccines
These studies show that PlpE is a surface exposed protein and was the only protein of 71 tested that was able to elicit a protective immune response
PlpE is not an essential virulence factor. This is the first report of a denatured recombinant protein stimulating protection against fowl cholera
Summary
Bacterial vaccines can be divided into three main categories: killed whole cell vaccines (bacterins), live attenuated vaccines and native or recombinant subunit vaccines. The concept of reverse vaccinology, first applied to Neisseria meningitidis [2], has been tested in three additional organisms [3,4,5] This method uses bioinformatics analysis of genomes to predict open reading frames (ORFs) encoding cell-surface or secreted proteins. These ORFs are expressed in a heterologous expression system and tested for their ability to elicit a protective immune response in animal models of infection. These works have expressed more than 800 proteins with 17 protective antigens identified; representing an identification rate of between 1 and 5%. We used a reverse vaccinology approach to identify putative secreted or cell surface associated P. multocida proteins that may represent potential vaccine candidate antigens
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
