Evaluation of a DNA vaccine encoding Brucella BvrR in BALB/c mice.

Brucellosis is an important neglected zoonotic disease, and the pathogens responsible are Brucellae. In order to evaluate the immunogenicity and protective efficacy of a DNA vaccine encoding Brucella BvrR, the recombinant plasmid pCDNA‑BvrR was constructed by inserting the BvrR gene fragment into a pCDNA3.0 vector. The His6‑tagged BvrR was purified with His‑trap FF crude affinity chromatography and verified with an anti‑histidine monoclonal antibody by western blot analysis. The specific immunoglobulin antigens and their isotypes were detected by indirect ELISA. The recombinant His6‑BvrR protein was expressed and purified by affinity chromatography. The optical density 450 value of immunoglobulin G (IgG) in the pCDNA‑BvrR group was significantly increased compared with the pCDNA3.0 vector or PBS groups (P<0.05), and the pCDNA3.0 vector and PBS groups exhibited no significant difference (P>0.05). BvrR induced specific antibodies with a dominance of IgG2a over IgG1 and the T cell‑proliferative response, in addition to a typical T helper‑1 (Th1)‑dominated immune response in mice. The splenocytes from mice of the pCDNA‑BvrR group demonstrated significant proliferative activity compared with the pCDNA3.0 vector group. The present results indicated that immunization with BvrR induced a specific Th1‑type immune response in mice. Subsequent to challenging with B. abortus S19, it was identified that the DNA vaccine pCDNA‑BvrR induced a significant level of protection in BALB/c mice by evaluating systemic bacterial clearance. These results suggested that BvrR may be a good candidate for a DNA vaccine against brucellosis.