Evaluation of recombinant Brachyspira pilosicoli oligopeptide-binding proteins as vaccine candidates in a mouse model of intestinal spirochaetosis.
The anaerobic intestinal spirochaete Brachyspira pilosicoli colonizes the large intestine of humans, and various species of animals and birds, in which it may induce a mild colitis and diarrhoea. The aim of the current study was to evaluate the use of putative oligopeptide-binding proteins of B. pilosicoli as vaccine components. A partial genome sequence of B. pilosicoli porcine strain 95/1000 was subjected to bioinformatics analysis, and six genes predicted to encode oligopeptide-binding proteins were selected. Following a PCR-based distribution study of the genes across different strains of the spirochaete, they were amplified from B. pilosicoli human strain WesB and cloned in Escherichia coli. The recombinant histidine-tagged proteins were purified and subjected to in vitro and in vivo immunogenicity analysis. Recombinant products (P-1 and P-3) from two genes that were immunogenic and recognized by sera from pigs that had recovered from B. pilosicoli infections were tested in a mouse model of intestinal spirochaetosis. For each recombinant protein, groups of 12 C3H/HeJ mice were vaccinated subcutaneously with 100 microg protein emulsified in Freund's incomplete adjuvant, twice with a 2 week interval. Two weeks later the vaccinated and non-vaccinated control animals were challenged orally with B. pilosicoli strain WesB. Both proteins induced systemic and local colonic IgG antibody responses, and, following experimental infection, the cumulative number of colonization days was significantly (P<0.001) less in both groups of vaccinated mice compared to the control mice. There were significantly (P=0.012) fewer mice colonized in the group vaccinated with P-1 than in the non-vaccinated control group. The results suggest that oligopeptide-binding proteins may have potential for use as components of vaccines for B. pilosicoli.
Movahedi
Abdolreza
A
Division of Health Sciences, School of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, Western Australia 6150, Australia.
Hampson
David J
DJ
Division of Health Sciences, School of Veterinary and Biomedical Sciences, Murdoch University, Murdoch, Western Australia 6150, Australia.
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology*; D017400:Brachyspira / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology*; D003092:Colitis / Q000382:microbiology / Q000517:prevention & control*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D003967:Diarrhea / Q000382:microbiology / Q000517:prevention & control*; D005243:Feces / Q000382:microbiology; D005260:Female; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology / Q000517:prevention & control*; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry / Q000276:immunology; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017422:Sequence Analysis, DNA; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D002352:Carrier Proteins; D004269:DNA, Bacterial; D008074:Lipoproteins; D011994:Recombinant Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; C046506:oligopeptide-binding protein, bacteria