Abstract

A DNA vaccine for use against Brucella maris was designed and developed in the Laboratory of Clinical Immunology and Microbiology (LCIM) at the U.S. Navy Marine Mammal Program (San Diego, CA). Gene targets were selected based on their role in pathogenesis and/or as known immunogens. Bacterial genes were amplified from RNA that was extracted from a clinical isolate of B. maris using RT-PCR with primers designed in-house. Subsequently, each gene was cloned into the commercially available pVax expression vector (Invitrogen, Carlsbad, CA), followed by sequence and expression analysis. Vaccine constructs will be tested initially in mice to confirm and characterize immunogenicity and immuno-kinetics. Correlates of protection for brucellosis indicate that the induction of balanced humoral and cellular immunity is required. Thus, this vaccine formulation has been designed to elicit both humoral and cellular immune responses. Immunogenicity will be determined by: i) the measurement of vaccine-specific antibodies, ii) demonstration of vaccine-specific cell-mediated immunity, and iii) cytokine analysis. Following successful demonstration of effectiveness in mice, the Brucella vaccine formulation will be evaluated for immunogenicity in a small group of Atlantic bottlenose dolphins (Tursiops truncatus).