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Infection and Immunity, November 2003, p. 6264-6269, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6264-6269.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evaluation of Brucella abortus Phosphoglucomutase (pgm) Mutant as a New Live Rough-Phenotype Vaccine

Juan Esteban Ugalde,1 Diego José Comerci,1,2 M. Susana Leguizamón,3 and Rodolfo Augusto Ugalde1*

Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín, Consejo Nacional de Investigaciones Científicas,1 Grupo Pecuario, Centro Atómico Ezeiza, Comisión Nacional de Energía Atómica,2 Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina3

Received 14 May 2003/ Returned for modification 27 June 2003/ Accepted 13 July 2003

Brucella abortus S19 is the vaccine most frequently used against bovine brucellosis. Although it induces good protection levels, it cannot be administered to pregnant cattle, revaccination is not advised due to interference in the discrimination between infected and vaccinated animals during immune-screening procedures, and the vaccine is virulent for humans. Due to these reasons, there is a continuous search for new bovine vaccine candidates that may confer protection levels comparable to those conferred by S19 but without its disadvantages. A previous study characterized the phenotype associated with the phosphoglucomutase (pgm) gene disruption in Brucella abortus S2308, as well as the possible role for the smooth lipopolysaccharide (LPS) in virulence and intracellular multiplication in HeLa cells (J. E. Ugalde, C. Czibener, M. F. Feldman, and R. A. Ugalde, Infect. Immun. 68:5716-5723, 2000). In this report, we analyze the protection, proliferative response, and cytokine production induced in BALB/c mice by a {Delta}pgm deletion strain. We show that this strain synthesizes O antigen with a size of approximately 45 kDa but is rough. This is due to the fact that the {Delta}pgm strain is unable to assemble the O side chain in the complete LPS. Vaccination with the {Delta}pgm strain induced protection levels comparable to those induced by S19 and generated a proliferative splenocyte response and a cytokine profile typical of a Th1 response. On the other hand, we were unable to detect a specific anti-O-antigen antibody response by using the fluorescence polarization assay. In view of these results, the possibility that the {Delta}pgm mutant could be used as a vaccination strain is discussed.

* Corresponding author. Mailing address: IIB-UNSAM, Av General Paz entre Constituyentes y Albarellos, PO Box 30 (1650) San Martín, Provincia de Buenos Aires, Argentina. Phone: 54-11-4580-7255/57. Fax: 54-11-4752-9639. E-mail: rugalde{at}iib.unsam.edu.ar.

Editor: W. A. Petri, Jr.

Infection and Immunity, November 2003, p. 6264-6269, Vol. 71, No. 11
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.11.6264-6269.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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