title: The immunogenicity of Rhodococcus equi GroEL2-based vaccines in a murine model.
abstract: Rhodococcus equi is a significant intracellular bacterial pathogen in foals. However, at present there is no commercially available vaccine for the prevention of R. equi-induced disease in these animals. Studies have shown that GroEL based vaccines can afford protection against some intracellular pathogens. In this study, the R. equi gene encoding the heat shock protein GroEL2 was cloned and sequenced, with a view to using it as a vaccine candidate. The promoter region of the gene contained two copies of controlling inverted repeat of chaperone expression (CIRCE) motifs, which are well-recognised transcriptional regulators of bacterial heat shock proteins. The R. equi GroEL2 was expressed in E. coli BL21 DE3 with a C-terminal His-tag and sequenced to confirm its identity. The R. equi purified His-tagged GroEL2 protein and a groEL2-based DNA vaccine were used in separate experiments to immunise BALB/c mice. The recombinant protein-based vaccine elicited a mixed Th1/Th2 response whereas the DNA vaccine was found to elicit a predominantly Th1 biased immune response. However, when vaccinated mice were challenged intravenously with 1.5 x 10(7) R. equi neither vaccine elicited enhanced bacterial clearance from the spleen or liver in this model. The reasons for this apparent lack of success are discussed.
authors.affiliation: School of Pharmaceutical, Molecular and Biomedical Sciences, University of South Australia, Adelaide, Australia.
authors.forename: Thiru
authors.initials: T
authors.lastname: Vanniasinkam
authors.forename: Mary D
authors.initials: MD
authors.lastname: Barton
authors.forename: Michael W
authors.initials: MW
authors.lastname: Heuzenroeder
mesh_terms: D000193:Actinomycetales Infections / Q000276:immunology / Q000517:prevention & control; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D001483:Base Sequence; D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005798:Genes, Bacterial; D006968:Hypersensitivity, Delayed; D051379:Mice; D008807:Mice, Inbred BALB C; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016929:Rhodococcus equi / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology / Q000494:pharmacology
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D018834:Chaperonin 60; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D019444:Vaccines, DNA; D022223:Vaccines, Subunit