Abstract
BackgroundHeat shock proteins (HSPs) are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune responses in mammals. We identified and characterized the hsp60-hsp10 bicistronic operon of the animal pathogen Corynebacterium pseudotuberculosis, a Gram-positive bacterium of the class Actinobacteria, which causes caseous lymphadenitis (CLA) in small ruminants.FindingsTo construct the DNA vaccine, the hsp60 gene of C. pseudotuberculosis was cloned in a mammalian expression vector. BALB/c mice were immunized by intramuscular injection with the recombinant plasmid (pVAX1/hsp60).ConclusionThis vaccination induced significant anti-hsp60 IgG, IgG1 and IgG2a isotype production. However, immunization with this DNA vaccine did not confer protective immunity.
Highlights
Heat shock proteins (HSPs) are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune responses in mammals
Isolation and characterization of the C. pseudotuberculosis hsp60 and hsp10 genes The C. pseudotuberculosis hsp60 gene was amplified by PCR using primers that were designed based on the genome of C. diphtheriae, because of the phylogenetic proximity between these two species
The nucleotide sequences of the C. pseudotuberculosis strain T1 60 kDa chaperonin and 10 kDa chaperonin GroES coding sequences were deposited in GenBank under accession numbers AY781285 and DQ869271, respectively
Summary
This vaccination induced significant anti-hsp IgG, IgG1 and IgG2a isotype production Immunization with this DNA vaccine did not confer protective immunity. Findings Corynebacterium pseudotuberculosis is a facultative, intracellular, Gram-positive bacterium of the class Actinobacteria, which includes the genera Mycobacterium, Nocardia and Rhodococcus. Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis (CLA), or cheesy gland, which affects small ruminants (sheep and goats) and occasionally other hosts. This chronic disease is pathognomonically characterized by the formation of suppurative abscesses in superficial and internal lymph nodes. Strategies to more effectively induce immunity with HSPs include the use of DNA vaccines. We assessed the feasibility of using DNA encoding hsp for protection against experimental challenge with C. pseudotuberculosis
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