The use of green fluorescent protein as a marker for Brucella vaccines

6 p., 3 figures, 1 table and referencesBrucellosis is an important malady of productive and wildlife animals and a worldwide zoonosis. The use of effective vaccines and the corresponding diagnostic tests that allow differentiating infected from vaccinated animals are essential tools to control the disease. For this, a prototype of Brucella abortus S19 vaccine expressing green fluorescent protein (S19-GFP) was constructed. The S19-GFP was readily identified under ultraviolet light by macroscopic and microscopic examination and maintained all the biochemical characteristics of the parental S19 vaccine. S19-GFP replicated ex vivo and in vivo, and protected mice against challenge with virulent B. abortus to the same extent as the isogenic S19. An immunoenzymatic assay designed to measure anti-GFP antibodies allowed the discrimination between mice vaccinated with S19-GFP and those immunized with S19. Both vaccines raised antibodies against lipopolysaccharide molecule to similar levels. This experimental model constitutes a “proof of concept” for the use of Brucella-GFP vaccines and associated diagnostic tests to distinguish vaccinated from naturally Brucella infected animals.This work was supported by NeTropica, Florida Ice and Farm, FIDA from Universidad Nacional, FS from CONARE, MICIT/CONICIT from Costa Rica and joint Costa Rica-Spain Bilateral Cooperation CRUSA-CSIC (2008CR0006).Peer reviewe

The use of green fluorescent protein as a marker for Brucella vaccines

La brucelosis es una importante enfermedad de los animales de producción y de la fauna silvestre y una zoonosis mundial. El uso de vacunas eficaces y las correspondientes pruebas de diagnóstico que permiten diferenciar los animales infectados de los vacunados son herramientas esenciales para controlar la enfermedad. Para ello se construyó un prototipo de vacuna de Brucella abortus S19 que expresa la proteína verde fluorescente (S19-GFP). La S19-GFP fue fácilmente identificada bajo luz ultravioleta por medio de un examen macroscópico y microscópico y mantuvo todas las características bioquímicas de la vacuna parental S19. La S19-GFP se replicó ex vivo e in vivo, y los ratones protegidos contra el desafío con la virulenta B. abortus en la misma medida que la isogénica S19. Un inmunoenzimático El ensayo diseñado para medir los anticuerpos anti-GFP permitió la discriminación entre los ratones vacunados con S19-GFP y los inmunizados con S19. Ambas vacunas elevaron los anticuerpos contra la molécula de lipopolisacárido a niveles similares. Este modelo experimental constituye una "prueba de concepto" para el uso de Vacunas de Brucella-GFP y pruebas diagnósticas asociadas para distinguir la vacuna de la Brucella natural. animales infectados.Brucellosis is an important malady of productive and wildlife animals and a worldwide zoonosis. The use of effective vaccines and the corresponding diagnostic tests that allow differentiating infected from vaccinated animals are essential tools to control the disease. For this, a prototype of Brucella abortus S19 vaccine expressing green fluorescent protein (S19-GFP) was constructed. The S19-GFP was readily identified under ultraviolet light by macroscopic and microscopic examination and maintained all the biochemical characteristics of the parental S19 vaccine. S19-GFP replicated ex vivo and in vivo, and protected mice against challenge with virulent B. abortus to the same extent as the isogenic S19. An immunoenzymatic assay designed to measure anti-GFP antibodies allowed the discrimination between mice vaccinated with S19-GFP and those immunized with S19. Both vaccines raised antibodies against lipopolysaccharide molecule to similar levels. This experimental model constitutes a “proof of concept” for the use of Brucella-GFP vaccines and associated diagnostic tests to distinguish vaccinated from naturally Brucella infected animals.Universidad Nacional, Costa RicaEscuela de Medicina Veterinari