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A DNA Vaccine Encoding Lumazine Synthase from Brucella abortus Induces Protective Immunity in BALB/c Mice

Facultad de Farmacia y Bioquímica Instituto de Estudios de la Inmunidad Humoral, and Facultad de Medicina,,¹ Laboratorio de Inmunogenética, Hospital de Clinicas "José de San Martin,",⁴ UBA, and Instituto de Investigaciones Bioquímicas (Fundación Campomar, IIBBA-CONICET, FCEN-UBA), Buenos Aires,² Laboratorio de Inmunoquímica y Biotecnología, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Ciencias Veterinarias, UNICEN, Tandil, Argentina³

Received 19 January 2001/ Returned for modification 12 April 2001/ Accepted 14 January 2002

This study was conducted to evaluate the immunogenicity of the Brucella abortus lumazine synthase (BLS) gene cloned into the pcDNA3 plasmid, which is driven by the cytomegalovirus promoter. Injection of plasmid DNA carrying the BLS gene (pcDNA-BLS) into BALB/c mice elicited both humoral and cellular immune responses. Antibodies to the encoded BLS included immunoglobulin G1 (IgG1) IgG2a, IgG2b, IgG3, and IgM isotypes. Animals injected with pcDNA-BLS exhibited a dominance of IgG2a over IgG1. In addition, spleen cells from vaccinated animals produced interleukin-2 and gamma interferon but not IL-10 or IL-4 after in vitro stimulation with recombinant BLS (rBLS), suggesting the induction of a Th1 response. Protection was evaluated by comparing the levels of infection in the spleens of vaccinated mice challenged with B. abortus 544. Immunization with pcDNA-BLS- reduced the bacterial burden relative to those in the control groups. Mice immunized with rBLS produced a significant humoral response but did not show a specific cellular response or any protection from challenge. Altogether, these data suggest that pcDNA-BLS is a good immunogen for the production of humoral and cell-mediated responses in mice and is a candidate for use in future studies of vaccination against brucellosis.

Editor: R. N. Moore

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